Non-glycosidic Iridoids from Cymbaria Mongolica

Phytochemistry. Mar, 2002  |  Pubmed ID: 11853749

Six non-glycosidic iridoids, i.e. (1R,4S,4aS,7S,7aS)-7-hydroxyl-4-hydroxy- methyl-7-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta[e]pyran-3-one (1), (1S,4R,4aS,7S,7aS)-7-hydroxyl-4-hydroxymethyl-7-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta[e]pyran-3-one (2), (1R,4R,4aS,7S,7aS)-7-hydroxyl-4-hydroxy-methyl-7-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta[e]pyran-3-one (3), (1R, 4R, 4aS, 7aS)-4,7-dihydroxymethyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta-6-ene[e]pyran-3-one (4), (1R, 4R, 4aS, 7aS)-4,7-dihydroxymethyl-1-hydroxyl-1,4,4a, 7a-tetrahydrocyclopenta-6-ene[e]pyran-3-one (5), (1R, 4S, 4aS, 7aS)-4,7-dihydroxy-methyl-1-methoxyl-1,4,4a,7a-tetrahydrocyclopenta-6-ene[e]pyran-3-one (6), as well as five known non-glycosidic iridoids mussaenin A (7), gardendiol (8), isoboonein (9), 4-epi-alyxialactone (10) and rehmaglutin D (11) have been isolated from the Chinese medicinal plant Cymbaria mongolica. Their structures were elucidated by spectroscopic methods. These compounds exhibit significant antitumor and antibacterial activity.

Graft Survival Assessment of MN Genotype After Allogeneic Transplantation of Hematopoietic Stem Cells of Umbilical Cord Blood

Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA. Oct, 2002  |  Pubmed ID: 12377617

To establish a simple and practical method to assess the outcome of allogeneic transplantation of hematopoietic stem cells from umbilical cord blood.

Antioxidant Effects of Green Tea Polyphenols on Free Radical Initiated Peroxidation of Rat Liver Microsomes

Chemistry and Physics of Lipids. Dec, 2002  |  Pubmed ID: 12426080

Antioxidative effects of the principal polyphenolic components extracted from green tea leaves, i.e. (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC), and gallic acid (GA), against free radical initiated peroxidation of rat liver microsomes were studied. The peroxidation was initiated by a water-soluble azo compound 2,2'-azobis(2-amidinopropane hydrochloride (AAPH). The reaction kinetics was monitored by oxygen uptake and formation of malondialdehyde (MDA). Kinetic analysis of the antioxidation process demonstrates that these green tea polyphenols (GOHs), especially EC and ECG which bear ortho-dihydroxyl functionality, are good antioxidants for microsomal peroxidation. The antioxidant synergism of these GOHs with the endogenous alpha-tocopherol (TOH) (vitamin E) is also discussed.

[Separation and Cryopreservation of Cord Blood Mononuclear Cells]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Aug, 2002  |  Pubmed ID: 12513772

The influencing factors on cord blood storage after collection and mononuclear cell separation as well as cryopreservation were studied. The mononuclear cell are separated from blood after blood collection, then cryopreserved and washed after thawed. Results showed that the cord blood kept at 4 degrees C or room temperature less than 24 hours after blood collection, mononuclear cell separated by hydroxyethylstarch and 2 centrifugations, mononuclear cell cryopreserved with 50% DMSO and autoplasma from cord blood as protectives and washing the cells after thawing. In conclusion, the optimal project in this study can effectively preserve cord blood mononuclear cells.

[Interspecies Nuclear Transfer Using Nonquiescent Somatic Cell]

Shi Yan Sheng Wu Xue Bao. Mar, 2002  |  Pubmed ID: 15344320

It is the point at issue in intraspecies nuclear transfer whether quiescence is necessary for development of nuclear transfer reconstructed embryos. In the interspecies nuclear transfer, some reports have proved that quiescent cell is able to support preimplantation development of the interspecies reconstructed embryos. Are non-quiescent cells able to support preimplantation development of the interspecies reconstructed embryos? We used non-quiescent somatic cells from C57BL/6 mice and giant pandas as donors to transfer into enucleated rabbit oocytes. After electrofusion (the electrofusion rates were 62.2% and 71.6%, respectively) and electrical activation, 5.1% of those mouse-rabbit reconstructed embryos developed to blastocyst in vitro, and 4.2% of panda-rabbit reconstructed embryos developed to blastocyst after transferring into ligated rabbit oviduct. These results indicate that non-quiescent cell from C57BL/6 mouse and giant panda could be dedifferentiated in enucleated rabbit oocytes and support early embryo development.

[Effects of Different Treatments on Oocytes Activation and Parthenogenesis in Mouse]

Shi Yan Sheng Wu Xue Bao. Sep, 2002  |  Pubmed ID: 15344387

In order to study effects of electro-fusion and strontium chloride (SrCl2) activation in nuclear transfer experiment on activation and development of mouse oocytes, concentration and treatment duration of SrCl2, electro-pulse and electro-pulse combining SrCl2 were used to activate mouse oocytes which were obtained after hCG 17h. Activated oocytes were cultured in vitro in CZB medium. The results were as follows: 82.4% activation percentage was obtained when the oocytes were treated with 10mmol/L SrCl2 for 6h, it was significantly (P>0.05) higher than those obtained from that treated with the 5mmol/L or 10mmol/L SrCl2 for 4h. The activation percentage was not significantly different between 5mmol/L and 10mmol/L SrCl2 for 6h, but the percentage of morula and blastocyst in 10mmol/L SrCl2 6h group was significantly (P > 0.05) higher than those in 5mmol/L SrCl2 6h group. In the groups of treatment with electro-pulse, the best activation percentage (70.9%) was obtained when the oocytes were treated with 1.0kv/cm, 320micros, 3 pulses, but M + B percentage (7.9%) was low. In the groups of treatment with electro-pulse combining with SrCl2, the best result was acquired (activation and M + B percentage were 75.0% and 57.3% separately) when the oocytes were treated in 10mmol/L SrCl2 for 6h interval 1h after treated with 1.8kv/cm, 10s, 1pulse. These results show that the treatment with electro-pulse combining SrCl2 is a better way to mouse parthenogenesis.

Inhibition of Free Radical-induced Peroxidation of Rat Liver Microsomes by Resveratrol and Its Analogues

Biochimica Et Biophysica Acta. Jan, 2003  |  Pubmed ID: 12527404

Resveratrol (3,5,4'-trans-trihydroxystilbene) is a natural phytoalexin present in grapes and red wine, which possesses a variety of biological activities including antioxidative activity. To find more efficient antioxidants by structural modification, resveratrol analogues, that is, 3,4-dihydroxy-trans-stilbene (3,4-DHS), 4,4'-dihydroxy-trans-stilbene (4,4'-DHS), 4-hydroxy-trans-stilbene (4-HS) and 3,5-dihydroxy-trans-stilbene (3,5-DHS), were synthesized and their antioxidant activity studied for the free radical-induced peroxidation of rat liver microsomes in vitro. The peroxidation was initiated by either a water-soluble azo compound 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH) or Fe(2+)/ascorbate, and monitored by oxygen uptake and formation of thiobarbituric acid reactive substances (TBARS). It was found that all of these trans-stilbene derivatives are effective antioxidants against both AAPH- and iron-induced peroxidation of rat liver microsomes with an activity sequence of 3,4-DHS>4,4'-DHS>resveratrol>4-HS>3,5-DHS. The remarkably higher antioxidant activity of 3,4-DHS is discussed.

Overexpression of ETS2 in Human Esophageal Squamous Cell Carcinoma

World Journal of Gastroenterology : WJG. Feb, 2003  |  Pubmed ID: 12532432

To study the expression pattern of ETS2 (erythroblastosis virus oncogene homolog 2) in human esophageal squamous cell carcinoma (ESCC).

Development of a Chinese Hamster Ovary Cell Line for Recombinant Adenovirus-mediated Gene Expression

Biotechnology Progress. Jan-Feb, 2003  |  Pubmed ID: 12573016

Recombinant human adenovirus (rhAd) has been used extensively for functional protein expression in mammalian cells including those of human and nonhuman origin. High-level protein production by rhAd vectors is expected in their permissive host cells, such as the human embryonic kidney 293 (HEK293) cell line. This is attributed primarily to the permissiveness of HEK293 to rhAd infection and their ability to support viral DNA replication by providing the missing El proteins. However, the HEK293 cells tend to suffer from cytopathic effect (CPE) as a result of virus replication. Under these circumstances, the host cell function is compromised and the culture viability will be reduced. Consequently, newly synthesized polypeptides may not be processed properly at posttranslational levels. Therefore, the usefulness of HEK293 cells for the expression of complex targets such as secreted proteins could be limited. In the search for a more robust cell line as a production host for rhAd expression vectors, a series of screening experiments was performed to isolate clones from Chinese hamster ovary-K1 (CHO-K1) cells. First, multiple rounds of infection of CHO-K1 cells were performed utilizing an rhAd expressing GFP. After each cycle of infection, a small population of CHO cells with high GFP levels was enriched by FACS. Second, individual clones more permissive to human adenovirus infection were isolated from the highly enriched subpopulation by serial dilution. A single clone, designated CHO-K1-C5, was found to be particularly permissive to rhAd infection than the parental pool and has served as a production host in the successful expression of several secreted proteins.

The Relation Between Apoptosis of Acinar Cells and Nitric Oxide During Acute Rejection of Pancreas Transplantation in Rats

Transplant Immunology. Jan-Mar, 2003  |  Pubmed ID: 12727471

Apoptosis is an important mechanism of immune-mediated graft damage. Nitric oxide (NO) generated by inducible NO synthase (iNOS) has been demonstrated to induce apoptosis. This study investigated whether apoptosis occurs during pancreas allograft rejection and examined the relationship of apoptosis of acinar cells and NO. The rats were divided into three groups: untreated isograft group, untreated allograft group and aminoguanidine (AG)-treated group. The pancreatic grafts were harvested on the post-transplantation day 3, 5 and 7 and were used to detect the histopathological rejection grade, the expression of iNOS and the apoptotic index (AI) of the graft. iNOS presented faint positive in the acinar cells of untreated isografts and did not change greatly after transplantation (P>0.05), the level of iNOS in the untreated allografts increased progressively (P<0.01) and at the same time point was significantly higher than that of untreated isograft group and AG-treated group (P<0.01). The transferase-mediated dUTP nick end labeling showed that the apoptotic cells were mainly acinar cells. A significant correlation between AI and iNOS was noted (P<0.01, r=0.611). Therefore, NO-mediated apoptosis of acinar cells plays an important role in acute rejection of pancreas transplantation, AG can mitigate the damage of pancreas allografts.

Neurotization of Oculomotor, Trochlear and Abducent Nerves in Skull Base Surgery

Chinese Medical Journal. Mar, 2003  |  Pubmed ID: 12781048

To anatomically reconstruct the oculomotor nerve, trochlear nerve, and abducent nerve by skull base surgery.

[ABO Blood Group Typing for Infants and Its Application for Clinical Transfusion]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Jun, 2003  |  Pubmed ID: 12844418

To study the correct method for determining ABO blood types in infants and its influencing factors, blood types of 33 infants under 6 months old were determined by routine serological method, micro-column gel typing system and PCR-SSP genotyping method. Of the 33 cases with discrepant results of ABO blood type by different methods, the blood types of 32 cases were discrepant between red cell and serological typings in the routine serological method, and a false coincidence in 1 case was caused by bacterial infection resulting in B-like antigen. Correct blood typing was obtained in 27 cases with a correct rate of 84.4% (27/32) by using micro-column gel typing system. PCR-SSP method gave correct results in all of 33 cases. There was a significant difference between the results of micro-column gel typing system and PCR-SSP. It is concluded that to determine ABO blood type for infants < 6 months old, it is recommended to adopt micro-column gel typing system method, and what must be taken into account is the possible false coincidence caused by bacterial infection resulting in B-like antigen. In micro-column gel typing system, if the results of red cell and serological typing are identical, the principle is that blood transfusion must be performed with same ABO blood type between recipient and donor. If not, washed O red blood cells should be used for infants, and then change to transfusion with identical blood group according to PCR-SSP typing results.

Pregnancy in a 46-year-old Woman After Autologous Granular Cell Mitochondria Transfer

Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA. Jul, 2003  |  Pubmed ID: 12865239

Growth of 4 follicles was achieved in a 46-year-old infertile woman and 2 mature oocytes were subsequently obtained after pituitary desensitization with triptorelin (1.875 mg) and hyperstimulation with recombinant human FSH (Gonal-F) and urinary human menopausal gonadotropin (HMG). The granular cells were collected from the cumulus and follicle fluid of the patient with Percol gradient centrifugation. The cells (about 1 x 10(6) /ml) were homogenized for 6 times with cold glass homogenizer, the resultant crude homogenate centrifuged for 20 min at 2000 x g. The supernatant was again centrifuged for 20 min at 9800 x g, after which the pellets of the mitochondria were resuspended in 1 microl modified human tubal fluid (mHTF) medium and maintained at 4 degrees Celsius. A microinjection needle with the inner diameter of approximately 7 microm was used to aspirate the mitochondrion suspension for about 100 microm in length on the scale of the needle for delivering the mitochondria, along with the immobilized sperms, into a single MII oocyte. About 3000 mitochondria were transferred into each oocyte. The two oocytes were both fertilized, but one had 3 pronucleus, and the normal fertilized oocyte developed into 8-blastomere embryo 3 d after its transfer into the patient's uterus. The urine test confirmed clinical pregnancy in the woman 15 d later, and 30 d after the transfer, the fetal body and heart beat could be observed by ultrasound, but unfortunately, spontaneous abortion occurred at the ninth week. According to our knowledge, this woman is the eldest person in mainland China to receive assisted reproduction operations that successfully got clinical pregnancy with her own oocyte.

[First Twins Born in Mainland China by Autologous Granular Cell Mitochondria Transfer]

Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA. Sep, 2003  |  Pubmed ID: 13129747

A 37-year-old woman with the history of twice spontaneous abortion, who received unfruitful in vitro fertilization and embryo transfer (IVF-ET) for two cycles due to poor embryo quality, underwent the third cycle for autologous granular cell mitochondria transfer of 5 matured oocytes and intracytoplasmic sperm injection (ICSI) in another 4 oocytes. Better embryo quality was resulted by the former technique than by the latter, despite that fertilization was achieved in all the 4 oocytes with the latter operation. Four of the 5 oocytes with mitochondria transfer were fertilized and developed into normal embryos, 3 of which were selected for intrauterine transplantation. Clinical pregnancy with triplets was subsequently obtained, but the development of one fetus accidentally stopped at the fifth week of pregnancy. The woman received five times of immunotherapy with her husband's leukocytes before and after the pregnancy. At the 30 th week of pregnancy, the women was admitted for severe pregnancy-induced hypertension syndrome that failed to respond to a two-week treatment course. Cesarean section was subsequently performed and two normal babies, one boy and one girl, were born on August 6th, 2003, who were taken home after care in neonate wards. This is the first report of the birth of babies born by autologous granular cell mitochondria transfer in Mainland China.

[Observation on Gene Polymorphism of Rh Blood Group in Chinese Han Nationality]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Dec, 2003  |  Pubmed ID: 14706151

To observe the gene polymorphism of Rh blood group in unrelated random individuals and families for Chinese Han nationality, polymerase chain reaction-sequence specific primer (PCR-SSP) was used to amplify the Rh C/E gene, RhD gene, exons, intron 2 and 10, insert and Rh Box in 160 blood samples of RhD positive unrelated individuals and 71 samples of RhD negative unrelated individuals and 7 samples of families whose probands were RhD-negative. The results showed that RhD genes of RhD-negative individuals with C antigens were polymorphism, three forms were found for D exon including intact, partial deletion and complete deletion exons. Insert fragments and Rh Box were found in most cases of families whose probands were RhD-negative and its inheritance accorded with the Mendel's Law, and it did not affect the expression of RhD gene. "Normal" RhD exon 4 amplifying product was not found in all of the samples. It was concluded that gene structure of the RhD-negative in Chinese was polymorphism, intact, partial deletion and complete deletion exons were found in the individuals with C antigen and probably existed specific D (nf) Ce haplotype. The function of insert was uncertain. The Rh gene sequences of Chinese Han nationality are different from those of Caucasian and the Rh gene library based on Han nationality should be established.

[A Clinical Study on Efficacy of Gemcitabine and Carboplatin Versus Paclitaxel and Carboplatin in the Treatment of Advanced Non-small Cell Lung Cancer.]

Zhongguo Fei Ai Za Zhi = Chinese Journal of Lung Cancer. Apr, 2003  |  Pubmed ID: 21262165

BACKGROUND: To evaluate the efficacy and toxicity of gemcitabine and carboplatin (GC) versus paclitaxel and carboplatin (TC) in the treatment of non-small cell lung cancer (NSCLC). METHODS: A total of 64 patients with advanced NSCLC diagnosed by pathology were randomly divided into two groups. Gemcitabine and carboplatin were administrated to the patients in GC group (n=30), and paclitaxel and carboplatin in the TC group (n=34), 28 days as a cycle. All patients received at least two cycles. RESULTS: The overall response rate was 53.3% in the GC group and 58.8% in the TC group (P > 0.05). The main toxicities were well tolerated and consisted of leukopenia, nausea, vomiting and peripheral neuritis, which occurred more frequently in the TC group than in the GC group (P < 0.05). CONCLUSIONS: Both of the two regimens of gemcitabine plus carboplatin and paclitaxel plus carboplatin are feasible, well tolerated and effective in the treatment of NSCLC, and the former may be safer than the latter.

Trapped Water of Human Erythrocytes and Its Application in Cryopreservation

Biophysical Chemistry. Feb, 2004  |  Pubmed ID: 14962599

The novel differential scanning calorimetry method as a technique for determining human red cell volume during freezing process has been reexamined and has been shown to provide a final erythrocyte volume to be 53% of its isotonic value after freezing from 0 to -40 degrees C. A new type of electronic particle counter (Multisizer 3, Beckman Coulter Inc., USA) was used to measure cell volume changes in response to equilibration in anisotonic media, and which gave out an equilibrated volume to be 57% of cell isotonic value in solution of 3186 mOsm. Both of these results indicate that 34-40% of intracellular water is trapped and is unavailable for participation in osmotic shifts. These findings are consistent with the published data that at least 20-32% (v/v) of the isotonic cell water is retained within RBCs. Then the application of trapped water in both simulation of freezing models and freezing-drying control was pointed out.

[Mitochondria Transfer from Self-granular Cells to Improve Embryos' Quality]

Zhonghua Fu Chan Ke Za Zhi. Feb, 2004  |  Pubmed ID: 15059588

To improve embryos' quality and pregnancy rate with the method of self-mitochondria transfer.

[Method of Detection of Soluble HLA-I and Soluble HLA-I Level Alteration in Storage Blood]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Jun, 2004  |  Pubmed ID: 15228668

Aim of this study was to develop the detection method of soluble human leukocyte antigens I (sHLA-I) and to explore sHLA-I level alteration in storage blood and its significance. sHLA-I level in sera of 60 Guangdong normal individuals and sHLA-I concentration in blood components from 20 donors quantitatively were detected by sandwich ELISA. The results showed that sensitivity of this assay was 2.84 ng/ml. Coefficients of variation were 5.80% within assays and 9.00% between assays respectively. The recovery rate was >/= 98.57%. The sHLA-I level of normal individuals in Guangdong was (699.54 +/- 360.10) ng/ml. sHLA-I in red blood cells stored for 28 days and in random-donor platelets were significantly higher than that in other blood components and their amount was proportionate to the number of residual donor leukocytes and to the length of storage. In conclusion, sandwich ELISA assay for detection of sHLA-I is a sensitive, specific and stable technique. Blood components with different concentration of sHLA-I may be chosen for clinical transfusion.

[Determination of L-cysteine in Enzymatic Reaction Mixture with Pre-column Derivatization by High Performance Liquid Chromatography]

Se Pu = Chinese Journal of Chromatography / Zhongguo Hua Xue Hui. May, 2004  |  Pubmed ID: 15712904

A new sensitive high performance liquid chromatographic method for the determination of L-cysteine in an enzymatic reaction mixture using ultra violet spectrometric detection was developed. The sample reacted with 5,5'-dithio-bis-nitrobenzoic acid (DTNB) and was analyzed on a Shimadzu VP-ODS column at room temperature, using gradient elution with detection at 330 nm. The L-cysteine chromatographic peak was determined in comparison with derivatives of 2-mercapto ethanol and dithiothreitol. The linear range was 5-950 micromol/L. The recoveries were 99.7%-100.5% and the relative standard deviations (RSDs) were less than 1.3%. The detection limit was 0.8 micromol/L. The method is simple and accurate.

Synergetic Inhibition of Metal Ions and Genistein on Alpha-glucosidase

FEBS Letters. Oct, 2004  |  Pubmed ID: 15474008

Inhibition of metal ions and synergetic inhibition of metal ions/genistein on alpha-glucosidase activity has been investigated. We have examined the inhibitory effect of Cu2+, Ni2+, Mg2+, Fe2+, Hg2+, Zn2+, Ca2+, Pb2+, Ag+, V5+, V4+ and Mn2+ ions. The results show that the nature of the inhibition was reversible, slow-binding, non-competitive, and the Ki values of some ions such as Cu2+, Ni2+ and Zn2+ range from 10(-5) to 10(-6) M. Moreover, synergetic inhibitory effect of metal ions and genistein on alpha-glucosidase were studied with kinetics method. It is concluded that the inhibitory effect was much greater when both of them were added to the reactant solution simultaneously than that they were added, respectively, which suggests that the inhibitors seem to bind to the different sites of alpha-glucosidase at the same time. Furthermore, the mechanism of the synergetic inhibition was examined by spectrophotometry.

Combined Use of Positive and Negative Immunomagnetic Isolation Followed by Real-time RT-PCR for Detection of the Circulating Tumor Cells in Patients with Colorectal Cancers

Journal of Molecular Medicine (Berlin, Germany). Nov, 2004  |  Pubmed ID: 15490093

To establish a novel molecular diagnostic method of detecting circulating tumor cells (CTCs) LS174T colon cancer cells were serially diluted with normal blood. Additional peripheral blood samples were collected from 25 patients with colorectal carcinoma. Mononuclear cells (MNCs) were collected, equally divided into four parts, and then cancer cells were enriched by four methods: method A, nonimmunobead method; method B, negative immunobead method: CD45 immunomagnetic beads were used to deplete the leukocytes; method C, positive immunobead method: Ber-EP4 immunomagnetic beads were used to enrich cancer cells; method D, negative-and-positive immunobead method: CD45 immunomagnetic beads were first used to deplete the leukocytes from MNC and then Ber-EP4 immunomagnetic beads were used to enrich cancer cells. Finally, real-time quantitative RT-PCR was used to monitor mRNA expression of beta2-mircoglobulin (beta2M) and carcinoembryonic antigen (CEA). The relative CEA mRNA values were corrected with reference to beta2M mRNA, to CEA mRNA/beta2M mRNA ratios according to a CEA mRNA external standards prepared with tenfold serial dilutions (1-10(4) IS174T cells) of cDNA and beta2M mRNA external standards prepared with tenfold serial dilutions (10(2)-10(7) leukocytes) of cDNA. In recovery experiments a significant correlation between the number of cancer cells and CEA mRNA expression was found when CD45 or Ber-EP4 immunomagnetic beads were used alone. A highly significant correlation was found when CD45 and Ber-EP4 immunomagnetic beads were used successively. The sensitivity of method D was one cancer cell per milliliter of blood. Circulating cancer cells were detected in 19 of 25 patients with colorectal cancers. The relative CEA mRNA value obtained by method D was the smallest. The positive detection rate of circulating cancer cells in patients at Dukes' B, C, and D stages were 25.0% (1/4), 83.3% (10/12), and 88.9% (8/9). Combinative use of immunomagnetic isolation followed by real-time RT-PCR is a useful technique to detect circulating tumor cells in patients with colorectal carcinomas. Applying negative and positive immunomagnetic beads successively yields the highest correlation with amount of tumor cells.

Trigeminal Neuralgia: What Are the Important Factors for Good Operative Outcomes with Microvascular Decompression

Surgical Neurology. Nov, 2004  |  Pubmed ID: 15518843

Microvascular decompression has been widely used as the first choice in treating trigeminal neuralgia, but in a few patients, facial pain cannot be effectively controlled by microvascular decompression. We sought to clarify the important factors for good operative outcomes.

Quantification of Sinomenine in Caulis Sinomenii Collected from Different Growing Regions and Wholesale Herbal Markets by a Modified HPLC Method

Biological & Pharmaceutical Bulletin. Jan, 2005  |  Pubmed ID: 15635172

Caulis Sinomenii is the dried plant stems of Sinomenium acutum and Sinomenium acutum var. cinereum and has been used in Chinese medicine for treating rheumatic diseases for over a thousand years. Previous studies have demonstrated that sinomenine is a major active constituent in both plants and can be utilized as an indicator of quality of the medicinal herb Caulis Sinomenii. Currently, S. acutum and S. acutum var. cinereum are growing over a wide geographical range in China, with equally wide variations in growing conditions. The objectives of this research were to determine whether there were difference between the species and varieties, and whether the different growing conditions could result in different quality by determining the content of sinomenine in different samples. A modified HPLC method using a diode array detector (DAD) has been developed for efficiently quantifying sinomenine in the plants. Using this method, fourteen samples of S. acutum var. cinereum and eleven samples of S. acutum from growing regions as well as eighteen herbal samples of Caulis Sinomenii from wholesale herbal markets were evaluated. The results showed that there was no marked difference in the content of sinomenine between the species and varieties collected from growing regions; however, a very large variation was found among the samples collected from different regions. Moreover, the content of sinomenine in the plants of large size (stem diameter>3 cm) was much higher than those of small size (stem diameter<1 cm). This implies that the growing region has greater impact on the quality of Caulis Sinomenii in terms of the content of sinomenine than the species and varieties. The results also showed that the content of sinomenine in commercial Caulis Sinomenii was markedly lower than that in the plants collected directly from growing regions. This suggests that to obtain the herb with higher content of sinomenine and thus ensure greater efficacy, both in clinical applications and in pharmacological investigations, the plant of Caulis Sinomenii with controlled stem size collected directly from growing regions is preferable.

High-throughput Mapping of a Dynamic Signaling Network in Mammalian Cells

Science (New York, N.Y.). Mar, 2005  |  Pubmed ID: 15761153

Signaling pathways transmit information through protein interaction networks that are dynamically regulated by complex extracellular cues. We developed LUMIER (for luminescence-based mammalian interactome mapping), an automated high-throughput technology, to map protein-protein interaction networks systematically in mammalian cells and applied it to the transforming growth factor-beta (TGFbeta) pathway. Analysis using self-organizing maps and k-means clustering identified links of the TGFbeta pathway to the p21-activated kinase (PAK) network, to the polarity complex, and to Occludin, a structural component of tight junctions. We show that Occludin regulates TGFbeta type I receptor localization for efficient TGFbeta-dependent dissolution of tight junctions during epithelial-to-mesenchymal transitions.

Influence of Co-administrated Sinomenine on Pharmacokinetic Fate of Paeoniflorin in Unrestrained Conscious Rats

Journal of Ethnopharmacology. May, 2005  |  Pubmed ID: 15848021

Paeonia lactiflora Pall. (Ranunculaceae) root and Sinomenium acutum Rehder and Wilson (Menispermaceae) stem are two herbs widely used in Chinese medicine to treat rheumatoid arthritis. While, in theory, either herb could be used alone, in practice, Chinese medicine practitioners prescribe them together. Studies on pharmacokinetic interaction between the active constituents of these two herbs (paeoniflorin and sinomenine, respectively) provide empirical evidence to support their clinical practice. A single dose of paeoniflorin (150 mg/kg) alone and with sinomenine hydrochloride (90 mg/kg) was administered by gastric gavage to unrestrained conscious male Sprague-Dawley rats (n=5 or 6, 240-270 [corrected] g). Blood samples were collected periodically via a jugular vein before and after dosing from 10 min to 12 h. A high-performance liquid chromatographic (HPLC) assay was developed to determine the plasma concentrations of paeoniflorin. Non-compartmental pharmacokinetic profiles were constructed by using the software PK Solutions 2.0. The pharmacokinetic parameters were compared using unpaired Student t-test. After co-administration of sinomenine, the peak plasma concentration of paeoniflorin was elevated (P<0.01), the peak time was delayed (P<0.01), the AUC(0-t) was increased (P<0.001), the mean residence time (MRT) was prolonged (P<0.01), the C(L) was decreased (P<0.01) and the V(d) was reduced (P<0.05). These results indicate that sinomenine hydrochloride at 90 mg/kg significantly improved the bioavailability of paeoniflorin in rats.

[Promotion of the Survival of Ischemic Skin Flap by Transplanted Endothelial Progenitor Cells Transfected with VEGF165 Gene: an Experimental Study with Mice]

Zhonghua Yi Xue Za Zhi. Feb, 2005  |  Pubmed ID: 15854554

To investigate the feasibility of transplanted endothelial progenitor cells transfected with VEGF165 gene to ischemic flap with increased neovascularization and augmented the survival areas.

Studies on the Operative Outcomes and Mechanisms of Microvascular Decompression in Treating Typical and Atypical Trigeminal Neuralgia

The Clinical Journal of Pain. Jul-Aug, 2005  |  Pubmed ID: 15951648

To evaluate the operative outcomes and mechanisms of microvascular decompression in treating typical and atypical trigeminal neuralgia.

Detecting AFP MRNA in Peripheral Blood of the Patients with Hepatocellular Carcinoma, Liver Cirrhosis and Hepatitis

Clinica Chimica Acta; International Journal of Clinical Chemistry. Nov, 2005  |  Pubmed ID: 15993394

The low frequency of disseminated carcinoma cells in the blood now makes immunomagnetic bead sorting and reverse transcriptase-polymerase chain reaction (RT-PCR) technique more popular.

Potent Antitumor Activity of Oncolytic Adenovirus Expressing Mda-7/IL-24 for Colorectal Cancer

Human Gene Therapy. Jul, 2005  |  Pubmed ID: 16000066

It has been demonstrated that interleukin 24 (IL-24, also called melanoma differentiation associated gene 7) exerts antitumor activity. In this study, we investigated whether oncolytic adenovirus-mediated gene transfer of IL-24 could induce strong antitumor activity. A tumor-selective replicating adenovirus expressing IL-24 (ZD55-IL-24) was constructed by insertion of an IL-24 expression cassette into the ZD55 vector, which is based on deletion of the adenoviral E1B 55-kDa gene. ZD55-IL-24 could express substantially more IL-24 than Ad-IL-24 because of replication of the vector. It has been shown that ZD55-IL-24 exerted a strong cytopathic effect and significant apoptosis in tumor cells with p53 dysfunction. Moreover, no cytotoxic and apoptotic effects could be seen in normal cells infected with ZD55-IL-24. Expression of IL-24 did not interfere with viral replication induced by oncolytic adenovirus. Activation of caspase 3 and caspase 9, and induction of bax gene expression, were involved in tumor cell apoptosis induced by ZD55-IL-24. Treatment of established tumors with ZD55-IL-24 showed much stronger antitumor activity than that induced by ONYX-015 or Ad-IL- 24. These data indicated that oncolytic adenovirus expressing IL-24 could exert potential antitumor activity and offer a novel approach to cancer therapy.

Detection of Cytokeratin 20 MRNA in the Peripheral Blood of Patients with Colorectal Cancer by Immunomagnetic Bead Enrichment and Real-time Reverse Transcriptase-polymeras Chain Reaction

Journal of Gastroenterology and Hepatology. Aug, 2005  |  Pubmed ID: 16048578

Detection of circulating cancer cells is a useful indicator for the risk of recurrence of advanced carcinoma. The aim of the present study was to evaluate the potential value of a novel approach to detect the circulating cancer cells in patients with colorectal cancer. This method is based on a combination of isolation of epithelial cell by a combination of negative and positive immunomagnetic beads with detection of cytokeratin 20 (CK20) mRNA by reverse transcriptase-polymeras chain reaction (RT-PCR).

A Study on the Sizes and Concentrations of Gold Nanoparticles by Spectra of Absorption, Resonance Rayleigh Scattering and Resonance Non-linear Scattering

Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy. Oct, 2005  |  Pubmed ID: 16165025

Liquid phase gold nanoparticles with different diameters and colors can be prepared using sodium citrate reduction method by controlling the amounts of sodium citrate. The mean diameters of gold nanoparticles are measured by transmission electron microscope (TEM). Gold nanoparticles with different sizes have specific absorption spectra. When the diameters of nanoparticles is between 12 and 41 nm, the maximum absorption peaks locate at 520-530 nm and there are red shifts gradually with the increase of diameters of gold nanoparticles. And when the size of gold nanoparticle is constant, the absorbance is proportional to the concentration of gold. Obvious resonance Rayleigh scattering (RRS) and the resonance non-linear scattering such as second-order scattering (SOS) and frequency-doubling scattering (FDS) appear at the same time as well, and the maximum scattering peaks are located at 286 nm (RRS), 480 nm (SOS) and 310 nm (FDS), respectively. When the concentration of gold is constant, absorbance and the intensities of RRS, SOS and FDS (I(RRS), I(SOS) and I(FDS)) have linear relationships with the diameters of gold nanoparticles. When the diameter of gold nanoparticle is constant, the absorbance and I(RRS), I(SOS), I(FDS) are directly proportional to the concentrations of gold nanoparticles. Therefore, it is very useful for studying the liquid phase gold nanoparticles by investigating the absorption, RRS, SOS and FDS spectra.

Suppressive Effects of QFGJS, a Preparation from an Anti-arthritic Herbal Formula, on Rat Experimental Adjuvant-induced Arthritis

Biochemical and Biophysical Research Communications. Nov, 2005  |  Pubmed ID: 16199007

To analyze the anti-arthritic effects of QFGJS (a pharmaceutical preparation from herbs) on rheumatoid arthritis, adjuvant-induced arthritis (AIA) was established in male SD rats, and two administration protocols, i.e., oral treatment with different doses of QFGJS on the day of arthritis induction or on the day when visible clinical signs of arthritis occurred, were initiated and continued until day 30. Treatments with QFGJS using both administration protocols significantly suppressed the incidence and severity of arthritis in a dose-dependent manner, showing dramatic reduction of paw swelling and ESR throughout the disease progression of AIA. Radiological and histopathological examinations showed markedly decreased tissue and bone destruction of ankle joints in the QFGJS-treated rats. The serum levels of TNF-alpha, IL-1beta, and IL-6 were significantly decreased in the QFGJS-treated rats. QFGJS demonstrates pronounced anti-arthritic effects on AIA, indicating that this herbal preparation would be a potent candidate as a novel botanical drug for further investigation.

Effects of Activin A on the Activities of the Mouse Peritoneal Macrophages

Cellular & Molecular Immunology. Feb, 2005  |  Pubmed ID: 16212913

Activin A is a kind of pre-inflammatory factor that belongs to the transforming growth factor-beta (TGF-beta) superfamily. To investigate the effect and mechanism of activin A on the activities of mouse macrophages, the secretion of NO in the supernatant of cultured mouse peritoneal macrophages was examined by NO assay kit, and the expression of iNOS, ActRIIA and ARIP2 mRNA in mouse peritoneal macrophages was analyzed by RT-PCR. The results showed that activin A stimulated the secretion of NO and the expression of iNOS mRNA in non-activated mouse macrophages in a time- and dose-dependent manner. In contrast, activin A in the same concentration inhibited the secretion of NO in LPS-activated mouse macrophages in a dose-dependent manner. ActRIIA was highly expressed on macrophages, and activin A upregulated the ActRIIA mRNA expression in macrophages. Anti-ActRIIA antibody can block the secretion of NO from the macrophages stimulated by activin A. Furthermore, RT-PCR analysis revealed that activin A enhanced the ARIP2 mRNA expression in macrophages. These results indicated that Activin A may be a weak activator compared with LPS to mouse macrophages, and activin A may modulate the secretion of NO through ActRIIA-ARIP2 signal pathway in mouse macrophages.

[Effects of Angiotensin II on Extracellular Signal-regulated Protein Kinases Signaling Pathway in Cultured Vascular Smooth Muscle Cells from Wistar-Kyoto Rats and Spontaneously Hypertensive Rats.]

Sheng Li Xue Bao : [Acta Physiologica Sinica]. Oct, 2005  |  Pubmed ID: 16220196

The aim of this study was to investigate the effects of angiotensin II (Ang II) on extracellular signal-regulated protein kinase (ERK) signaling pathway in cultured vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. VSMCs from SHR and WKY rats were treated with 1x10(-7) mmol/L Ang II for 24 h in the absence or presence of 30 min of pre-treatment of valsartan (1x10(-5) mmol/L) or PD98059 (1x10(-5)mmol/L), selective inhibitor of ERKs- dependent pathways, when they were cultured in 20% calf serum medium. VSMCs of SHR and WKY cultured in serum-free medium were used as control groups. Among the different treatments, VSMCs from the SHR and WKY were devided into four groups: (1) control, (2) Ang II, (3) Ang II + valsartan, (4) Ang II + PD98059. ERK activity in VSMCs was measured by immuno-precipitation. Proteins of total ERK (t-ERK), phosphorylated-ERK (p-ERK) and mitogen-activated protein kinases phosphatase-1 (MKP-1) in VSMCs were detected by Western blot. MKP-1 mRNA in VSMCs was measured by RT-PCR. In VSMCs from WKY or SHR rats, ERK activity, p-ERK, MKP-1 and MKP-1 mRNA in Ang II group were higher than those in control group (P<0.05). In both SHRs and WKYs, there were no significant differences in ERK activity, p-ERK, MKP-1 and MKP-1 mRNA among the control group, Ang II + valsartan group and Ang II + PD98059 group. ERK activity, p-ERK, MKP-1 and MKP-1 mRNA in SHRs were significantly higher than those in WKYs with same treatments (P<0.01). There was no significant difference in t-ERK among different groups and no difference in t-ERK between SHRs and WKYs (P>0.05). Our results show that Ang II activates VSMCs ERK signaling pathways via Ang II type 1 (AT(1)) receptors. Ang II increased ERK activity and p-ERK, but not t-ERK, accompanied by an increase in MKP-1 mRNA expression and protein. Among the different treatments, ERK activity and p-ERK were higher in SHR than in WKY. Valsartan and PD98059 blocked Ang II-stimulated ERK activation. These results suggest that ERK signaling pathway plays an important role in the pathogenesis of hypertension. The effect of Ang II on SHR and WKY VSMCs' ERK signaling pathway may be mediated by AT(1) receptors, enhancing ERK activity and the amount of p-ERK, and then increasing MKP-1 mRNA and its expression.

Pharmacokinetic Interaction of Paeoniflorin and Sinomenine: Pharmacokinetic Parameters and Tissue Distribution Characteristics in Rats and Protein Binding Ability in Vitro

Journal of Pharmacological Sciences. Dec, 2005  |  Pubmed ID: 16327213

The root of Paeonia lactiflora and the stem of Sinomenium acutum are two herbs widely used in Chinese herbal medicine for the treatment of inflammatory and arthritic diseases. Studies on the interaction of the active constituents of these herbs, i.e., paeoniflorin and sinomenine, in pharmacokinetic parameters, tissues distribution, and protein binding ability could provide empirical data to support their clinical application. Following oral administration to rats, the pharmacokinetic alterations were compared. The results showed that the pharmacokinetic parameters (Cmax, Tmax, AUC, MRT, C(L), and Vd) of paeoniflorin were markedly enhanced when co-administrated with sinomenine. At 45 min after oral administration, the concentrations of paeoniflorin in the main internal organs were significantly increased when co-administrated with sinomenine. These phenomena were not ascribable to the alteration of the protein binding ability of paeoniflorin by sinomenine because obvious interactions of paeoniflorin and sinomenine in protein binding abilities in vitro to rat and rabbit plasma, human albumin, and alpha-1-acid-glycoprotein were not observed. However, with respect to the in vivo influence of paeoniflorin on sinomenine, the results showed that co-administration of paeoniflorin did not affect the pharmacokinetic parameters and tissue distribution of sinomenine.

[ERK Expression and Activation in Myocardial Hypertrophy of Spontaneously Hypertensive Rats with Different Ages]

Zhejiang Da Xue Xue Bao. Yi Xue Ban = Journal of Zhejiang University. Medical Sciences. Nov, 2005  |  Pubmed ID: 16331818

To investigate the expression and activation of extracellular signal-regulated kinases (ERK) in myocardial hypertrophy of spontaneously hypertensive rats (SHR) with different ages.

Suppressive Effects of JCICM-6, the Extract of an Anti-arthritic Herbal Formula, on the Experimental Inflammatory and Nociceptive Models in Rodents

Biological & Pharmaceutical Bulletin. Feb, 2006  |  Pubmed ID: 16462027

JCICM-6, the extract of an anti-arthritic herbal formula composed of medicinal herbs of Sinomenium acutum, Aconitum carmichaeli DEBX., Curcuma Longa L., Paeonia lactiflora PALL., and Paeonia suffruticosa ANDR., was examined in the effectiveness and mechanism in reducing experimentally-induced inflammation and nociception using nine animal models. JCICM-6 was extracted from herbs and purified with Amberlite XAD-7HP adsorbent resin and analyzed with HPLC-fingerprint for quality consistency. In acute inflammatory models, the paw edema of rats was induced by subcutaneous injection of carrageenan or pro-inflammatory mediators, including histamine, serotonin, bradykinin, and prostaglandin E(2) (PGE(2)) into the right hind paws of animals; while the ear edema of mice was induced by applying arachidonic acid or 12-O-tetradecanoylphorbol 13-acetate (TPA) on the ear surface. In nociceptive models, the tail-flick response induced by radiant heat stimulation was measured and the numbers of abdominal writhing episodes of mice induced by intraperitoneal injection of acetic acid were recorded. JCICM-6 orally administered in a range of dosages from 0.438 g to 1.75 g/kg significantly and dose-dependently suppressed the paw edema of rats induced by carrageenan or various pro-inflammatory mediators and the ear edema of mice induced by arachidonic acid or TPA. JCICM-6 also significantly prolonged the reaction time of rats to radiant heat stimulation and reduced the numbers of writhing episodes of mice. These results indicated that JCICM-6 possesses significant anti-inflammatory and analgesic effects, which implies that it would be a potential candidate for further investigation as a new anti-arthritic botanical drug for humans.

Curcumin Analogs As Potent Aldose Reductase Inhibitors

Archiv Der Pharmazie. Mar, 2006  |  Pubmed ID: 16528793

In the present study, curcuminoids isolated from curcuma longa were demonstrated to possess inhibitory activities on bovine lens aldose reductase. In order to find more potent aldose reductase inhibitor, curcumin analogs were synthesized and evaluated for their ability to inhibit bovine lens aldose reductase enzyme. The results indicated that the compounds with tetrahydroxyl groups, 2,6-bis(3,4-dihydroxybenzylidene)cyclohexanone (A(2)), 2,5-bis(3,4-dihydroxybenzylidene)cyclopentanone (B(2)), 1,5-bis(3,4-dihydroxyphenyl)-1,4-pentadiene-3-one (C(2)), and 3,5-bis(3,4-dihydroxybenzylidene)-4-piperidone (D(2)) showed remarkably potent inhibitory effects on aldose reductase with IC(50) of 2.9 microM, 2.6 microM, 3.4 microM, and 4.9 microM, respectively. The structure-activity relationship revealed that the curcumin analogs with ortho-dihydroxyl groups could form a more tight affinity with aldose reductase to exert more potential inhibitory activities.

Beta-2 Adrenergic Receptor Polymorphisms and the Forearm Blood Flow Response to Mental Stress

Clinical Autonomic Research : Official Journal of the Clinical Autonomic Research Society. Apr, 2006  |  Pubmed ID: 16683069

Circulating epinephrine plays an important role in skeletal muscle vasodilation during mental stress. Normotensive adults homozygous for glycine (Gly) of the Arg16/Gly beta2-adrenergic receptor polymorphism have a greater forearm beta2-receptor mediated vasodilation and a higher cardiac output response to isometric handgrip than arginine (Arg) homozygotes. To test the hypothesis that the Arg16/Gly beta2-adrenergic receptor polymorphism affects the forearm blood flow (FBF) and hemodynamic response to mental stress, and whether venous catecholamine concentrations predicted these responses, we measured venous epinephrine, norepinephrine, heart rate (HR), arterial pressure (Finapres), and FBF during mental stress in healthy subjects homozygous for Gly16 (n = 30; mean age +/- SE: 30 +/- 1.2, 13 women) and Arg16 (n = 17, age 30 +/- 1.6, 11 women). Resting HR, blood pressure, and FBF responses to mental stress were similar between genotype groups. There were positive correlations between epinephrine and peak FBF (r = 0.694, P < 0.001), peak forearm vascular conductance (r = 0.677, P < 0.001) and the change in epinephrine to the change in HR (r = 0.456, P = 0.002) in all subjects. These correlations were not significantly different in the Gly16 and Arg16 groups. We conclude that venous epinephrine predicts the FBF response to mental stress, and the increase in epinephrine is also correlated with the increase in HR. Furthermore, the Arg16/Gly beta2-receptor polymorphism has no significant influence on the FBF or cardiovascular responses to mental stress.

Analysis of Thrombotic Factors in Severe Acute Respiratory Syndrome (SARS) Patients

Thrombosis and Haemostasis. Jul, 2006  |  Pubmed ID: 16807662

Systemic Hypoxia and Vasoconstrictor Responsiveness in Exercising Human Muscle

Journal of Applied Physiology (Bethesda, Md. : 1985). Nov, 2006  |  Pubmed ID: 16809628

Exercise blunts sympathetic alpha-adrenergic vasoconstriction (functional sympatholysis). We hypothesized that sympatholysis would be augmented during hypoxic exercise compared with exercise alone. Fourteen subjects were monitored with ECG and pulse oximetry. Brachial artery and antecubital vein catheters were placed in the nondominant (exercising) arm. Subjects breathed hypoxic gas to titrate arterial O2 saturation to 80% while remaining normocapnic via a rebreath system. Baseline and two 8-min bouts of rhythmic forearm exercise (10 and 20% of maximum) were performed during normoxia and hypoxia. Forearm blood flow, blood pressure, heart rate, minute ventilation, and end-tidal CO2 were measured at rest and during exercise. Vasoconstrictor responsiveness was determined by responses to intra-arterial tyramine during the final 3 min of rest and each exercise bout. Heart rate was higher during hypoxia (P < 0.01), whereas blood pressure was similar (P = 0.84). Hypoxic exercise potentiated minute ventilation compared with normoxic exercise (P < 0.01). Forearm blood flow was higher during hypoxia compared with normoxia at rest (85 +/- 9 vs. 66 +/- 7 ml/min), at 10% exercise (276 +/- 33 vs. 217 +/- 27 ml/min), and at 20% exercise (464 +/- 32 vs. 386 +/- 28 ml/min; P < 0.01). Arterial epinephrine was higher during hypoxia (P < 0.01); however, venoarterial norepinephrine difference was similar between hypoxia and normoxia before (P = 0.47) and during tyramine administration (P = 0.14). Vasoconstriction to tyramine (%decrease from pretyramine values) was blunted in a dose-dependent manner with increasing exercise intensity (P < 0.01). Interestingly, vasoconstrictor responsiveness tended to be greater (P = 0.06) at rest (-37 +/- 6% vs. -33 +/- 6%), at 10% exercise (-27 +/- 5 vs. -22 +/- 4%), and at 20% exercise (-22 +/- 5 vs. -14 +/- 4%) between hypoxia and normoxia, respectively. Thus sympatholysis is not augmented by moderate hypoxia nor does it contribute to the increased blood flow during hypoxic exercise.

[In Vivo Transduction of EGFP into Female Mouse Reproductive System by Electroporation and Microbubble-enhanced Sonoporation]

Fen Zi Xi Bao Sheng Wu Xue Bao = Journal of Molecular Cell Biology / Zhongguo Xi Bao Sheng Wu Xue Xue Hui Zhu Ban. Aug, 2006  |  Pubmed ID: 16955797

In the present study, the efficiency of in vivo transduction of EGFP plasmid DNA into adult female mouse reproductive system by means of electroporation and microbubble-enhanced sonoporation are discussed. In the first experiment,EGFP plasmid DNA was injected into the ovary and following electroporation (square electric pulses were applied five times at 60V with a constant time of 20ms). Green fluorescence was detected 10h after transduction. In the second experiment, plasmid DNA was mixed with the microbubble at different ratio and injected into the uterus and the ovary separately. The tissues were exposed to different combined ultrasound parameters immediately after injection. We found that the suitable parameter was 2 w/cm2 intensity, 20% duty cycle and 5min. Under this condition, green fluorescence was expressed in the uterus endometrium and ovarian cortex 12 h post-sonoporation and the expression lasted up to 5 days.

Cloning, Expression, and Identification of Genes Involved in the Conversion of DL-2-amino-Delta2-thiazoline-4-carboxylic Acid to L-cysteine Via S-carbamyl-L-cysteine Pathway in Pseudomonas Sp. TS1138

Bioscience, Biotechnology, and Biochemistry. Sep, 2006  |  Pubmed ID: 16960371

Two novel genes (tsB, tsC) involved in the conversion of DL-2-amino-Delta2-thiazoline-4-carboxylic acid (DL-ATC) to L-cysteine through S-carbamyl-L-cysteine (L-SCC) pathway were cloned from the genomic DNA library of Pseudomonas sp. TS1138. The recombinant proteins of these two genes were expressed in Escherichia coli BL21, and their enzymatic activity assays were performed in vitro. It was found that the tsB gene encoded an L-ATC hydrolase, which catalyzed the conversion of L-ATC to L-SCC, while the tsC gene encoded an L-SCC amidohydrolase, which showed the catalytic ability to convert L-SCC to L-cysteine. These results suggest that tsB and tsC play important roles in the L-SCC pathway and L-cysteine biosynthesis in Pseudomonas sp. TS1138, and that they have potential applications in the industrial production of L-cysteine.

The Effects of Sinomenine on Intestinal Absorption of Paeoniflorin by the Everted Rat Gut Sac Model

Journal of Ethnopharmacology. Feb, 2006  |  Pubmed ID: 16169700

Paeoniflorin and sinomenine, derived from the root of Paeonia lactiflora Pall. (family Ranunculaceae) and the stem of Sinomenium acutum Rehder & Wilson (family Menispermaceae), respectively, have been, and are currently, widely used for treatment of rheumatic and arthritic diseases in China and Japan. Our previous studies demonstrated that sinomenine could significantly improve the bioavailability of paeoniflorin in rats, but the underlying mechanisms remain unknown. The present study aims to investigate the intestinal kinetic absorptive characteristics of paeoniflorin as well as the absorptive behavior influenced by co-administration of sinomenine using an in vitro everted rat gut sac model. The results showed a good linear correlation between the paeoniflorin absorption in sac contents and the incubation time from 0 to 90 min. However, the concentration dependence showed that a non-linear correlation exists between the paeoniflorin absorption and its concentrations from 10 to 160 microM, and the absorption was saturated at about 80 microM of the drug. Sinomenine at 16 and 136 microM concentrations could significantly enhance the absorption of paeoniflorin (20 microM) by 1.5- and 2.5-fold, respectively. Moreover, two well-known P-glycoprotein inhibitors, verapamil and quinidine, could significantly elevate the absorption of paeoniflorin by 2.1- and 1.5-fold, respectively. Furthermore, sinomenine in a pattern, which influenced paeoniflorin's absorption, manifested as similar to that of P-glycoprotein inhibitors. In conclusion, sinomenine significantly enhance the intestinal absorption of paeoniflorin, subsequently improve the bioavailability of paeoniflorin. The mechanism underlying the improvement of paeoniflorin's bioavailability was proposed that sinomenine could decrease the efflux transport of paeoniflorin by P-glycoprotein.

Mechanisms Responsible for Poor Oral Bioavailability of Paeoniflorin: Role of Intestinal Disposition and Interactions with Sinomenine

Pharmaceutical Research. Dec, 2006  |  Pubmed ID: 17063398

To determine the intestinal disposition mechanisms of paeoniflorin, a bioactive glucoside, and to investigate the mechanisms by which sinomenine increases paeoniflorin bioavailability.

Resonance Rayleigh Scattering Study on the Interaction of Gold Nanoparticles with Berberine Hydrochloride and Its Analytical Application

Analytica Chimica Acta. Jul, 2006  |  Pubmed ID: 17723490

The interaction of gold nanoparticles with berberine hydrochloride has been studied by using resonance Rayleigh scattering (RRS) spectra. In pH 3.8-5.5 aqueous solution, citrate acid ([H2L2-]) self-assembled on the surface of positively charged gold nanoparticles (average diameter is about 12.0 nm) to form a supermolecular complex with negative charges. By virtue of electrostatic attraction, hydrophobic force and charge transfer, the complex bound with berberine to form complex, which had bigger diameter (35 nm) than gold nanoparticles. The formation of the binding production not only resulted in the red shift of absorption of gold nanoparticles from 518 to 672 nm, but also led to the greatly enhancement of RRS intensity. At the same time, the intensities of second-order scattering (SOS) and frequency-doubling scattering (FDS) were also increased. Under definite condition, the increment of the RRS (DeltaI) were proportional to the concentration of berberine. A sensitive and simple method for the determination of berberine based on the RRS technique has been developed. The detection limit (3sigma) for berberine was 0.40 ng mL(-1) and the quantitative determination range was 1.33-240 ng mL(-1). In this work, the optimum conditions of reaction, the effect of foreign substances and the analytical application had been investigated.

Enhanced Plasmon Resonance Light Scattering Signals of Colloidal Gold Resulted from Its Interactions with Organic Small Molecules Using Captopril As an Example

Analytica Chimica Acta. Sep, 2006  |  Pubmed ID: 17723679

Gold nanoparticles are known for their plasmon resonance absorption (PRA) depending on their size. Our this investigation shows that plasma resonance light scattering (PRLS) signals in the corresponding PRA region could be measured using a common spectrofluorometer, and be enhanced when aggregation of gold nanoparticles occurs due to their interaction with organic small molecules (OSMs). Using captopril (Cap) as an example, we investigated the interactions of gold nanoparticles with OSMs in order to propose a general method of OSMs such as typical clinic organic drugs. In aqueous medium of pH 2.09, there are about 2.2 x 10(3) Cap molecules covalently binding to the surface of a 10-nm diameter gold nanoparticle through the thiol functional group of Cap, and thus forms a core-shell assembly of [(Au)(31000)]@[(Cap)(2200)], displaying strong enhanced PRLS signals in the PRA region of gold colloid. The PRLS intensities characterized at 553.0 nm were found to be proportional to the concentration of Cap over the range of 0.1-1.7 mg L(-1) with the determination limit (3sigma) of 32.0 microg L(-1). With that, Cap in pharmaceutical preparations could be determined with the recovery of 97.0-104.5% and R.S.D. of less than 2.4%.

Light Scattering Sensing Detection of Pathogens Based on the Molecular Recognition of Immunoglobulin with Cell Wall-associated Protein A

Analytica Chimica Acta. Sep, 2007  |  Pubmed ID: 17870291

In this contribution, we report a rapid optical detection method of pathogens using Staphylococcus aureus (S. aureus) as the model analyte based on the molecular recognition of immunoglobulin with cell wall-associated Protein A (SpA). It was found that the molecular recognition of human immunoglobulin (IgG) with protein A on the cell wall of S. aureus on glass slide sensing area could result in strong surface enhanced light scattering (SELS) signals, and the SELS intensity (deltaI) increases proportionally with the concentration of S. aureus over the range of 2.5x10(5)-1.0x10(8) CFU mL(-1) with right angle light scattering (RALS) signals detection mode. In order to identify the solid support based molecular recognition between IgG with SpA, we also employed water-soluble CdS quantum dots (CdS-QDs) as a fluorescent marker for IgG by immobilizing the IgG onto the surfaces of CdS-QDs through covalent binding in order to generate recognition probes for SpA on the cell wall of S. aureus. Consequently, the fluorescent method also showed that the detection for pathogens with solid supports is reliable based on the molecular recognition of IgG with SpA.

Baroreflex Sensitivity Inversely Correlates with Ambulatory Blood Pressure in Healthy Normotensive Humans

Hypertension. Jul, 2007  |  Pubmed ID: 17502489

Patients with hypertension have a blunted sensitivity of baroreflex control of heart period. In these patients, baroreflex sensitivity is positively related to heart rate variability and inversely related to blood pressure variability. We hypothesized that this relationship would also be evident in healthy normotensive subjects and that individuals with higher baroreflex sensitivity would have lower ambulatory 24-hour blood pressure. Twenty-four-hour ambulatory blood pressure and heart rate were recorded in 50 healthy, normotensive, nonobese individuals (31 women and 19 men). The baroreflex was assessed using sequential bolus administration of sodium nitroprusside and phenylephrine, and baroreflex sensitivity was calculated as the slope of the relation between systolic blood pressure and R-R interval during the resulting blood pressure transients. Baroreflex sensitivity was inversely correlated to 24-hour average mean arterial pressure (R=0.49; P<0.001) and positively related to daytime heart rate variability (R=0.33; P=0.02). In contrast, no relationship was found between baroreflex sensitivity and 24-hour heart rate or blood pressure variabilities. We conclude that the relationship between baroreflex sensitivity and daytime heart rate variability was similar to that reported previously in hypertensive subjects. Furthermore, the inverse relation between baroreflex sensitivity and mean arterial pressure supports the idea that the baroreflex may exert longer-term effects on blood pressure than thought previously.

Workshop Report on the Extraction of Foetal DNA from Maternal Plasma

Prenatal Diagnosis. Sep, 2007  |  Pubmed ID: 17604339

Cell free foetal DNA (cff DNA) extracted from maternal plasma is now recognized as a potential source for prenatal diagnosis but the methodology is currently not well standardized. To evaluate different manual and automated DNA extraction methods with a view to developing standards, an International Workshop was performed.

Detecting Carcinoma Cells in Peripheral Blood of Patients with Hepatocellular Carcinoma by Immunomagnetic Beads and Rt-PCR

Journal of Clinical Gastroenterology. Sep, 2007  |  Pubmed ID: 17700428

Increasing the sensitivity and specificity of detecting circulating carcinoma cells of patients with hepatocellular carcinoma (HCC) is very important for monitoring recurrence.

Balance Between Sympathetic Response to Head-up Tilt and Cardiac Vagal Factors in Healthy Humans

Clinical Autonomic Research : Official Journal of the Clinical Autonomic Research Society. Aug, 2007  |  Pubmed ID: 17717719

We evaluated the association between cardio-vagal baroreflex sensitivity (BRS; assessed with modified Oxford technique) and catecholamine response to 5 min 60 degrees head-up tilt (HUT) in 46 young healthy adults. HUT increased HR, mean arterial pressure, and NE (P < 0.05 for all). BRS was negatively correlated with NE response to HUT (r = -0.36, P < 0.05), suggesting that subjects with high vagal modulation (high BRS) require less sympathetic response (NE) to maintain normotension during orthostatic stress.

Resonance Rayleigh Scattering Spectral Method for the Determination of Raloxifene Using Gold Nanoparticle As a Probe

Analytica Chimica Acta. Aug, 2007  |  Pubmed ID: 17719906

When gold nanoparticles were being prepared by sodium citrate reduction method, citrate anions self-assembled on the surface of gold nanoparticles to form supermolecular complex anions with negative charges, and protonated raloxifene (Ralo) was positively charged and could bind with the complex anions to form larger aggregates through electrostatic force and hydrophobic effects, which could result in the remarkable enhancement of the resonance Rayleigh scattering intensity (RRS), and the appearance of new RRS spectra. At the same time, the second-order scattering (SOS) and frequency-doubling scattering (FDS) intensities were also enhanced. The maximum wavelengths were located near 370 nm for RRS, 520 nm for SOS, and 350 nm for FDS, respectively. Among them, the RRS method had the highest sensitivity and the detection limit was 5.60 ng mL(-1) for Ralo, and its linear range was 0.05-2.37 microg mL(-1). A new RRS method for the determination of trace Ralo using gold nanoparticles probe was developed. The optimum conditions of the reaction and influencing factors were investigated. In addition, the reaction mechanism and the reasons for the enhancement of RRS were discussed.

[Cloning, Antisense Construction and Tomato Transformation of Lecop1like Gene]

Fen Zi Xi Bao Sheng Wu Xue Bao = Journal of Molecular Cell Biology / Zhongguo Xi Bao Sheng Wu Xue Xue Hui Zhu Ban. Oct, 2007  |  Pubmed ID: 18254338

Here reported a 1060 bp cDNA cloning of LeCOP1LIKE gene by EST probing and RT-PCR method. In order to characterize function of this gene, a LeCOP1LIKE antisense expression vector was transformed into Micro-Tom via Agrobacterium-mediated transformation method and 10 independent transgenic lines were obtained. RT-PCR analysis showed that the expression of LeCOP1LIKE gene was evidently repressed in 4 lines of them. The transgenic plants were much shorter than their wild type control, their chlorophyll content was increased but the seed development was obviously suppressed. All these results suggested that the cloned LeCOP1LIKE gene was a negative regulator of photomorphogenesis in tomato.

[Application of Metabolomics in Research of Plant Metabolites]

Zhongguo Zhong Yao Za Zhi = Zhongguo Zhongyao Zazhi = China Journal of Chinese Materia Medica. Nov, 2007  |  Pubmed ID: 18257252

Metabolomics, a branch of systems biology, has gained extensive attention and profound achievements in the plant. Although plant metabolomics is to be explored, it has been one of the most effective methods to study the physiological and biochemical process and gene modification in pattern plants. We herein summarized the concept, development, and application of metabolomics and prospected the potentials in the metabolite profiling for plant. Metabolomics provides an omics' methodology to elucidate the whole biological process, identify and quantify the complex components in the plant. A number of metabolites present in the plant are active components of traditional Chinese medicine, and these bioactive components are influenced by the multi-factors such as environment, species, and processing methods etc. Therefore, it is of great importance to analyze a wide spectrum of compositions with diverse chemical characteristics and varied concentration, which is the foundation to quality control, allowing the elucidation of the pharmacological effectiveness, and further exploiting of traditional Chinese medicine.

Suppression of the Onset and Progression of Collagen-induced Arthritis in Rats by QFGJS, a Preparation from an Anti-arthritic Chinese Herbal Formula

Journal of Ethnopharmacology. Mar, 2007  |  Pubmed ID: 17049776

QFGJS is an herbal preparation, and its pronounced effectiveness in treating adjuvant-induced arthritis (AIA) has been previously demonstrated. We herein aimed to confirm its anti-arthritic effect on collagen-induced arthritis (CIA) in rats. CIA was established in female Wistar rats with intradermal injection of type II bovine collagen at the base of the tail of animals. CIA rats were treated daily with oral administration of different doses of QFGJS beginning on the day of the induction of arthritis (day 0, the prophylactic treatment) or on the day after the onset of arthritis (day 13, the therapeutic treatment) until day 30. The results showed that prophylactic treatment with QFGJS significantly suppressed the onset of arthritis, and therapeutic treatment with QFGJS markedly reduced paw swelling and ESR levels even in the established CIA. Radiologic and histopathologic changes in the arthritic joints were also significantly reduced in the QFGJS-treated versus vehicle-treated rats. Moreover, the serum levels of pro-inflammatory cytokines TNF-alpha, IL-1beta, and IL-6 were markedly lowered in the QFGJS-treated rats. Hence, our studies demonstrate the quality, safety, and effectiveness of QFGJS as an anti-arthritic agent, which makes QFGJS a strong candidate for further clinical trials on rheumatoid arthritis (RA) patients.

Ultra-performance LC/TOF MS Analysis of Medicinal Panax Herbs for Metabolomic Research

Journal of Separation Science. Apr, 2008  |  Pubmed ID: 18338405

In this study, metabolite profiling of five medicinal Panax herbs including Panax ginseng (Chinese ginseng), Panax notoginseng (Sanchi), Panax japonicus (Rhizoma Panacis Majoris), Panax quinquefolium L. (American ginseng), and P. ginseng (Korean ginseng) were performed using ultra-performance LC-quadrupole TOF MS (UPLC-QTOFMS) and multivariate statistical analysis technique. Principal component analysis (PCA) of the analytical data showed that the five Panax herbs could be separated into five different groups of phytochemicals. The chemical markers such as ginsenoside Rf, 20(S)-pseudoginsenoside F11, malonyl gisenoside Rb1, and gisenoside Rb2 accountable for such variations were identified through the loadings plot of PCA, and were identified tentatively by the accurate mass of TOFMS and partially verified by the available reference standards. Results from this study indicate that the proposed method is reliable for the rapid analysis of a group of metabolites present in herbal medicines and other natural products and applicable in the differentiation of complex samples that share similar chemical ingredients.

Fetal DNA: Strategies for Optimal Recovery

Methods in Molecular Biology (Clifton, N.J.). 2008  |  Pubmed ID: 18425483

For fetal DNA extraction, in principle each DNA extraction method can be used; however, because most methods have been optimized for genomic DNA from leucocytes, we describe here the methods that have been optimized for the extraction of fetal DNA from maternal plasma and validated for this purpose in our laboratories. The use of the QIAamp DSP Virus kit (QIAGEN), the QIAamp DNA Blood Mini kit (QIAGEN), and the Magna Pure LC (Roche) is based on the kit components provided by the respective companies. However, we noticed that the yield of fetal DNA from maternal plasma can be increased when higher volumes are processed or some slight modifications of the protocols provided by the manufacturer are followed. Here, we also describe an in-house method that allows the specific capture of target molecules in an extremely low volume by using magnetic beads and magnetic tips. This method can be either performed by hand, or it can be adapted to a commercially available pipetting workstation.

[Production of Porcine Reconstructed Embryos by Whole-cell Intracytoplasmic Microinjection]

Fen Zi Xi Bao Sheng Wu Xue Bao = Journal of Molecular Cell Biology / Zhongguo Xi Bao Sheng Wu Xue Xue Hui Zhu Ban. Feb, 2008  |  Pubmed ID: 18464592

Cloning by somatic cell nuclear transfer has been achieved by both electric fusion and intracytoplasmic nuclear injection (ICNI) methods. However, each of the above methods involves extended complicate manipulation and special equipment. Here we report a whole-cell injection technique without Piezo assistance for nuclear transfer in pigs. The fibroblast cell of pig as the nucleus donor cell, effects of the new method on the efficiency of somatic nuclear transfer in pig were investigated, compared with that of electric fusion method. Results showed that the new method was a little less efficient in producing reconstructed embryos but without significant difference (88.4% vs 78%, P > 0.05). After the embryos were cultured 48h and 7d, the fusion method is more efficient than the new method in the oocyte cleavage rate and the blastocyst development (78% vs 53.2%, P < 0.05; 27.2% vs 13.8%, P < 0.01). The results indicate that both methods make no difference in the quality of the blastula, but the electric fusion method is more efficient. Therefore, the applicability of producing normal,cloned piglets by the simple and less labor-intensive whole-cell intracytoplasmic injection needs further improvement.

Efficient Production of Mice from Embryonic Stem Cells Injected into Four- or Eight-cell Embryos by Piezo Micromanipulation

Stem Cells (Dayton, Ohio). Jul, 2008  |  Pubmed ID: 18467666

The conventional method for producing embryonic stem (ES) cell-derived knockout or transgenic mice involves injection of ES cells into normal, diploid blastocysts followed by several rounds of breeding of resultant chimeras and thus is a time-consuming and inefficient procedure. F0 ES cell pups can also be derived directly from tetraploid embryo complementation, which requires fusion of two-cell embryos. Recently, F0 ES cell pups have been produced by injection of ES cells into eight-cell embryos using a laser-assisted micromanipulation system. We report a simple method for producing F0 ES cell germline-competent mice by piezo injection of ES cells into four- or eight-cell embryos. The efficiency of producing live, transgenic mice by this method is higher than that with the tetraploid blastocyst complementation method. This efficient and economical technique for directly producing F0 ES cell offspring can be applicable in many laboratories for creating genetically manipulated mice using ES cell technology and also for stringent testing of the developmental potency of new ES cell or other types of pluripotent stem cell lines.

Metabolite Profiling of Panax Notoginseng Using UPLC-ESI-MS

Phytochemistry. Aug, 2008  |  Pubmed ID: 18550132

The metabolite profiling of different parts of Panax notoginseng was carried out using rapid ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) and multivariate statistical analysis. Principal component analysis (PCA) of the UPLC-ESI-MS data showed a clear separation of compositions among the flower buds, roots and rhizomes of P. notoginseng. The saponins accounting for such variations were identified through the corresponding loadings weights and were further verified by accurate mass, tandem mass and retention times of available standard saponins using UPLC quadrupole time-of-flight mass spectrometer (UPLC-QtofMS). Finally, the influential factors of different metabolic phenotypes of P. notoginseng was elucidated. The currently proposed UPLC-ESI-MS/MS analytical method coupled with multivariate statistical analysis can be further utilized to evaluate chemical components obtained from different parts of the plant and/or the plant of different geographical locations, thereby classifying the medicinal plant resources and potentially elucidating the mechanism of inherent phytochemical diversity.

A Novel Mutation in EXT2 Gene in a Chinese Family with Hereditary Multiple Exostoses

Genetic Testing. Sep, 2008  |  Pubmed ID: 18666861

Hereditary multiple exostoses (HME) is an autosomal-dominant disorder characterized by the presence of bony outgrowths on the long bones. In this report, we describe a Chinese family with HME. Linkage analysis and mutation detection were performed. Linkage with the EXT2 was established in this family. A novel mutation, EXT2 c239-244delG, was identified. Mutation analysis in a family with HME allows for genetic counseling and prenatal diagnosis.

[Allogeneic Leukocyte Immunization Combined with IVF-ET for Treatment of Infertility Induced by Recurrent Spontaneous Abortion]

Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University. Jul, 2008  |  Pubmed ID: 18676284

To investigate the effect of allogeneic leukocyte immunization combined with in vitro fertilization-embryo transfer (IVF-ET) for treatment of infertility induced by habitual abortion.

Synthesis and Evaluation of Curcumin Analogues As Potential Thioredoxin Reductase Inhibitors

Bioorganic & Medicinal Chemistry. Sep, 2008  |  Pubmed ID: 18678491

Series of curcumin derivatives were synthesized; the inhibitory activities on thioredoxin reductase (TrxR) of all analogues were evaluated by DTNB assay in vitro. It is found that most of the analogues can inhibit TrxR in the low micromolar range; Structure-activity relationship analysis reveals that analogues with furan moiety have excellent inhibitory effect on TrxR in an irreversible manner, indicating that the furan moiety may serve as a possible pharmacophore during the interaction of curcumin analogues with TrxR. The effect of selected curcuminoids on growth of different TrxR overexpressed cancer cell lines was also investigated and discussed.

Inhibition of Thioredoxin Reductase by Curcumin Analogs

Bioscience, Biotechnology, and Biochemistry. Aug, 2008  |  Pubmed ID: 18685195

Curcumin analogs were first investigated for their inhibitory effects on thioredoxin reductase (TrxR). Most of them were more potent TrxR inhibitors than natural curcumin. The structure-activity relationship was summarized, and the curcumin analog was found to inhibit TrxR irreversibly in a time-dependent manner. The action was caused by covalent modification of the redox-active residues Cys(497) and Sec(498) in TrxR.

Cardiovascular Dynamics in Healthy Subjects with Differing Heart Rate Responses to Tilt

Journal of Applied Physiology (Bethesda, Md. : 1985). Nov, 2008  |  Pubmed ID: 18756006

Orthostatic stress such as head-up tilt (HUT) elicits a wide range of heart rate (HR) and arterial pressure (AP) responses among healthy individuals. In this study, we evaluated cardiovascular dynamics in healthy subjects with different HR responses to HUT, but without autonomic dysfunction. We measured AP (brachial artery) and HR (ECG) during 5 min of 60 degrees HUT in 76 healthy normotensive individuals. We then chose individuals on the basis of the extremes of HR responses to HUT (high = DeltaHR > or = 20 beats/min, and low = DeltaHR < or = 10 beats/min; n = 15 per group). Peak HR during HUT was 87 +/- 10 beats/min in the high and 69 +/- 14 beats/min in the low group (P < 0.05). High HR responders had lower systolic pressure at baseline (121 +/- 9 vs. 129 +/- 11 mmHg, P < 0.05) and during HUT (120 +/- 10 vs. 131 +/- 13 mmHg, P < 0.05), and higher plasma norepinephrine (NE) response to HUT (DeltaNE: 156.9 +/- 17.8 vs. 89.0 +/- 17.2 pg/ml; P < 0.05). DeltaNE during HUT was also significantly correlated with DeltaHR when all 76 subjects were included in a regression analysis (r = 0.39; P < 0.001). Pulse pressure was lower during HUT in high HR responders compared with low HR responders (45 +/- 1 vs. 55 +/- 2 mmHg, P < 0.05). High HR responders also had larger fluctuations in systolic and pulse pressure during HUT (coefficient of variation = 10.7 +/- 0.7 vs. 5.7 +/- 0.3%; 7.9 +/- 0.5 vs. 4.1 +/- 0.4%, respectively, P < 0.05). Sex distribution was different between groups (high: 5 women, 10 men; low: 10 women, 5 men). Higher HR with lower AP during HUT is consistent with normal baroreflex mechanisms of integration. Although interindividual variability appears to be a fundamental part of cardiovascular regulation, the mechanisms of these differences and the sex discrepancy requires further investigation.

[Culture of Porcine Mesenchymal Stem Cells in Vitro and Developmental Capacity of Reconstructed Embryos by Nuclear Transfer]

Fen Zi Xi Bao Sheng Wu Xue Bao = Journal of Molecular Cell Biology / Zhongguo Xi Bao Sheng Wu Xue Xue Hui Zhu Ban. Aug, 2008  |  Pubmed ID: 18959001

The developmental potential of reconstructed embryos varied according to the source of donor cells, it was thought that the donor cells capabilities to be reprogrammed were different. We established the method of culturing porcine bone marrow mesenchymal stem cells (pMSCs), identified and observed the growth characteristics of pMSCs, and determined pMSCs reprogramming potential as donor cells for nuclear transfer (SCNT). We found that the method of gradient centrifugation to isolate pMSCs from porcine bone marrow was better than the method of anchoring culture; the number of pMSCs achieved peak at day 6, the adhesive rate of cultured cells was 78.50% at 10h and the division index of cultured cells was 24.00 per thousand at day 4. The developmental competence were compared among three kinds of embryos, reconstructed embryos with PF and pMSCs, Parthenogenetic. The blastocysts rate and total cell number of blastocysts were 15.07%, 14.63% vs 30.91% and 24.1 +/- 6.5, 30.67 +/- 17.7 vs 25.8 +/- 11.4. These results indicated that pMSCs could be high proliferation and stable growth characters in vitro, and were suitable donor cells type for nuclear transfer.

[Construction and Identification of the Eukaryotic Expression Vector PIRES2-EGFP-IL-1ra-Fcepsilon]

Fen Zi Xi Bao Sheng Wu Xue Bao = Journal of Molecular Cell Biology / Zhongguo Xi Bao Sheng Wu Xue Xue Hui Zhu Ban. Aug, 2008  |  Pubmed ID: 18959005

The cDNA of IgE constant domain of rat was cloned from the spleen of allergy asthma rat by RT-PCR. The IL-1ra segment was obtained from intermediate vector pBV220-IL-1ra. By overlap extension PCR, the fusion gene IL-1ra-Fcepsilon was cloned, then inserted into the eukaryotic expression plasmid pIRES2-EGFP to obtain a recombinant expression plasmid pIRES2-EGFP-IL-1ra-Fcepsilon. The recombinant expression plasmid was transfected into 293T cells using lipofectamin and instillated into the rat lung through trachea. The expression of IL-1ra-Fcepsilon was identified by Western blot, RT-PCR, and this protein could inhibit the activity of IL-1 in vitro. Green fluorescent protein could be detected in the transfected 293T cells and the rat lungs at different times. The research paved the way for the gene therapy of allergy asthma.

Studies on the Phenylethanoid Glycosides with Anti-complement Activity from Paulownia Tomentosa Var. Tomentosa Wood

Journal of Asian Natural Products Research. Nov-Dec, 2008  |  Pubmed ID: 19031237

Four epimeric phenylethanoid glycosides, including a new one, R,S-beta-ethoxy-beta-(3,4-dihydroxyphenyl)-ethyl-O-alpha-L-rhamnopyranosyl(1-->3)-beta-D-(6-O-E-caffeoyl)-glucopyranoside named isoilicifolioside A (1), and three known compounds, ilicifolioside A (2), campneoside II (3), and isocampneoside II (4), were isolated from Paulownia tomentosa var. tomentosa wood. The structures of the four compounds were elucidated by the interpretation of 1D and 2D NMR and MS spectra. This is the first report of the chemical profile of this tree. Compounds 1-4 exhibited excellent anti-complement activity with IC(50) values less than 74 microM, compared with tiliroside (IC(50) = 104 microM) and rosmarinic acid (IC(50) = 182 microM) that were used as positive controls.

Body Iron Stores and Dietary Iron Intake in Relation to Diabetes in Adults in North China

Diabetes Care. Feb, 2008  |  Pubmed ID: 17909092

To evaluate the association between body iron stores, dietary iron intake, and risk of diabetes in northern China.

A Localized Surface Plasmon Resonance Light-scattering Assay of Mercury (II) on the Basis of Hg(2+)-DNA Complex Induced Aggregation of Gold Nanoparticles

Environmental Science & Technology. Jul, 2009  |  Pubmed ID: 19673301

It is known that localized surface plasmon resonance (LSPR) is responsible for the surface-enhanced spectroscopic processes of metallic nanoparticles and thus LSPR spectroscopy has become a powerful technique for chemical and biological purposes. In this contribution, we present a simple homogeneous Hg2+ assay by measuring enhanced LSPR scattering signals resulted from Hg(2+)-DNA complex induced aggregation of gold nanoparticles (AuNPs). In a medium of pH 7.4 tris-HCl buffer containing 0.05 M NaCl, single-stranded oligonucletides with the sequence of 5'-d(T6)-3' (poly-T6 ssDNA), can be selectively adsorbed onto the surface of gold colloids, stabilizing the AuNPs against aggregation. If Hg(2+)-DNA complex via Hg(2+)-mediated thymine-Hg(2+)-thymine (T-Hg(2+)-T) is formed, however, the adsorption of poly-T6 ssDNA onto the surface of gold colloids gets reduced, and then aggregation of the AuNPs occurs owing to the decrease of the electrostatic repulsion between AuNPs. Consequently, strong LSPR scattering signals resulting from the aggregates of AuNPs could be visually observed under a dark field microscope and easily be measured with a common spectrofluorometer. The LSPR scattering intensities characterized at 556.0 nm were found to be proportional to the concentration of Hg2+ ions in the range of 4.0 x 10(-8) to 6.0 x 10(-7) M with the limit of determination (3sigma) of 1.0 nM. Compared with reported colorimetric methods, our present approaches display the advantages of higher sensitivity.

Characterization of High-yield Pulp (HYP) by the Solute Exclusion Technique

Bioresource Technology. Dec, 2009  |  Pubmed ID: 19692226

By following the solute exclusion technique, we determined the pore characteristics of 3 hardwood high-yield pulps (aspen, birch, maple). The fiber saturation point (FSP) was 1.40, 1.36 and 1.19 g water/g pulp, for aspen, birch, maple, respectively, which is lower than that of chemical pulps. Different fractions obtained from the Bauer-McNett classifier showed that the HYP fines have much more pore volume than their long fiber counterparts. The effects of beating, drying and re-wetting on the pore characteristics of HYP were also studied. Beating led to increased total pore volumes. Upon drying and re-wetting, much of the small pores from the HYP underwent permanent closure while the big pores were only slightly affected. Finally, the relationship between the water retention value (WRV) and FSP for HYP was examined.

[Recent Advances in Treatment of Glial Scar-induced Inhibition of Nerve Regeneration]

Zhejiang Da Xue Xue Bao. Yi Xue Ban = Journal of Zhejiang University. Medical Sciences. Nov, 2009  |  Pubmed ID: 20014492

Glial scar formed by central nervous system (CNS) injury is the main inhibitory barrier of nerve regeneration. How to promote axonal regeneration after injury,how to accelerate neural network reconstruction and how to improve brain function recovery have become a hot problem to be solved in the field of neuroscience. This article focuses on the recent advances of therapeutic strategies for axonal regeneration.

[VEGF and Central Nervous System Diseases]

Zhejiang Da Xue Xue Bao. Yi Xue Ban = Journal of Zhejiang University. Medical Sciences. Nov, 2009  |  Pubmed ID: 20014493

Vascular endothelial growth factor (VEGF or VEGF-A) is a hypoxia induced angiogenic growth factor that is potent in neurotrophy,neuroprotection, anti-apoptosis and cell proliferation. Recent reports suggest that VEGF is related to many central nervous system diseases, such as cerebral ischemic disease, Alzheimer's disease and Parkinson's disease. Further study of the relationship between VEGF and central nervous system diseases,and investigation of VEGF related drugs will shed light on a new way for treatment of central nervous system diseases.

Resveratrol Ameliorates Carbon Tetrachloride-induced Acute Liver Injury in Mice

Environmental Toxicology and Pharmacology. Nov, 2009  |  Pubmed ID: 21784026

Present investigation aimed to evaluate the hepatoprotective potential of resveratrol (30mg/kg, po) in mice following two different routes (po and sc) of exposure to carbon tetrachloride (CCl(4), 1.0ml/kg). Administration of CCl(4) caused significant increase in the release of transaminases, alkaline phosphatase, lactate dehydrogenase, γ-glutamyl transpeptidase, creatinine kinase, total bilirubin, urea and uric acid in serum. Significantly enhanced hepatic lipid peroxidation and oxidized glutathione with marked depletion in reduced glutathione were observed after CCl(4) intoxication. It was also found that CCl(4) administration caused severe alterations in liver histology. Hepatic injury was more severe in those animals who received CCl(4) by oral route than those who exposed to CCl(4) subcutaneously. Resveratrol treatment was able to mitigate hepatic damage induced by acute intoxication of CCl(4) and showed pronounced curative effect against lipid peroxidation and deviated serum enzymatic variables as well as maintained glutathione status toward control. Treatment of resveratrol lessened CCl(4) induced damage in liver. The results of the present study suggest that resveratrol has potential to exert curative effects against liver injury.

Active Control of a 30 M Ring Interferometric Telescope Primary Mirror

Applied Optics. Feb, 2009  |  Pubmed ID: 19183591

The active control of a primary mirror for a ring aperture segmented telescope is different from that of a full aperture segmented telescope. Two active maintenance proposals for the ring telescope designed from the segmented patterns are outlined. We present a preliminary calculation of noise propagation and analysis of primary mirror mode characteristics for each proposal. The modulation transfer functions (MTFs) of the primary mirror corresponding to each maintenance method at several typical working wavelengths are also simulated.

Upper Cervical Spine Chordoma of C2-C3

European Spine Journal : Official Publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society. Mar, 2009  |  Pubmed ID: 19221808

Chordoma is a rare low-grade malignant neoplasm derived from the remnants of the embryonic notochord. This locally invasive neoplasm is subject to recurrence after treatment. The median survival time is estimated to be 6.3 years. Various treatment approaches have been attempted, including radical excision, radiotherapy and chemotherapy. Treatment outcome is significantly influenced by the size and site of the chordoma. Recently, Imatinib, a molecular-targeted agent, has been shown to have antitumor activity in chordoma. Proton radiotherapy, stereotactic radiotherapy and intensity-modulated radiotherapy have also been used. Surgical treatment is still the primary choice for chordoma. It has become more aggressive in recent years, evolving from intralesional or partial excision to en bloc resection. However, upper cervical localizations make such en bloc resection in most cases not possible. We present and discuss the therapeutic challenges of a young female with large retropharyngeal chordoma who presented to our institution after conventional photon beam radiotherapy. This C2/3 tumor was classified IB according to the Enneking classification. It distributed to layers A-D and sectors 1-6 according to the Weinstein Boriani Biagini Classification. The left vertebral artery (VA) was encapsulated and displaced. One stage intralesional extracapsular tumor excision and reconstruction was achieved by combined bilateral high anterior cervical approaches and posterior approach. No recurrence or metastasis was observed 3 years after the operation. She returned to her previous occupation as office worker.

Correlation of Expression and Methylation of Imprinted Genes with Pluripotency of Parthenogenetic Embryonic Stem Cells

Human Molecular Genetics. Jun, 2009  |  Pubmed ID: 19324901

Mammalian parthenogenetic embryos (pE) are not viable due to placental deficiency, presumably resulting from lack of paternally expressed imprinted genes. Pluripotent parthenogenetic embryonic stem (pES) cells derived from pE could advance regenerative medicine by avoiding immuno-rejection and ethical roadblocks. We attempted to explore the epigenetic status of imprinted genes in the generation of pES cells from parthenogenetic blastocysts, and its relationship to pluripotency of pES cells. Pluripotency was evaluated for developmental and differentiation potential in vivo, based on contributions of pES cells to chimeras and development to day 9.5 of pES fetuses complemented by tetraploid embryos (TEC). Consistently, pE and fetuses failed to express paternally expressed imprinted genes, but pES cells expressed those genes in a pattern resembling that of fertilized embryos (fE) and fertilized embryonic stem (fES) cells derived from fE. Like fE and fES cells, but unlike pE or fetuses, pES cells and pES cell-fetuses complemented by TEC exhibited balanced methylation of Snrpn, Peg1 and U2af1-rs1. Coincidently, global methylation increased in pE but decreased in pES cells, further suggesting dramatic epigenetic reprogramming occurred during isolation and culture of pES cells. Moreover, we identified decreased methylation of Igf2r, Snrpn, and especially U2af1-rs1, in association with increased contributions of pES cells to chimeras. Our data show that in vitro culture changes epigenetic status of imprinted genes during isolation of pES cells from their progenitor embryos and that increased expression of U2af1-rs1 and Snrpn and decreased expression of Igf2r correlate with pluripotency of pES cells.

Isolation and Culture of Primary Bovine Embryonic Stem Cell Colonies by a Novel Method

Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology. Jun, 2009  |  Pubmed ID: 19340839

Authentic bovine embryonic stem (ES) cell lines have not been established despite progress made for more than two decades. Isolation and culture of primary ES cell colonies are the first critical step towards establishment of stable ES cell lines. Here we report a novel method designated as "Separate and Seed" that contributes remarkably to efficient derivation of bovine primary ES-like cell colonies from blastocysts. These primary cultured bovine ES-like cells exhibit morphology typical of ES cells and express pluripotent molecular markers including Oct4, Nanog and alkaline phosphatase. Interestingly, bovine primary ES-like cell colonies distinctively express both stage-specific embryonic antigens 1 and 4 (SSEA1 and SSEA4), unlike mouse and human ES cells. These pluripotent markers may be used for characterization of authentic bovine ES cell lines in later studies. In contrast, whole embryos or inner cell mass (ICM) used for primary culture by conventional methods fails to produce primary bovine ES cell colonies that express all pluripotent stem cell markers shown above. Furthermore, bFGF improves growth and maintained undifferentiated state of bovine ES-like cells for several passages, whereas LIF and ERK inhibitor PD98059 known to promote pluripotency of mouse ES cells are unable to sustain bovine ES-like cells. Although continued efforts are required for improving long-term culture of bovine ES cells, this novel "Separate and Seed" method provides an initial effective step that may eventually lead to derivation of authentic bovine ES cell lines.

Specific Magnetic Bead-based Capture of Free Fetal DNA from Maternal Plasma

Transfusion and Apheresis Science : Official Journal of the World Apheresis Association : Official Journal of the European Society for Haemapheresis. Jun, 2009  |  Pubmed ID: 19364676

An automated magnetic capture hybridization (MCH) method for the extraction and enrichment of fetal RHD specific DNA fragments from maternal plasma was developed using plasma from 1000 D-negative pregnant women. A real time PCR protocol for RHD exon 7 was applied. MCH was compared with the QIAamp DSP Virus Kit (QIAamp) as a reference. Compared with the QIAamp method, the percentage of fetal DNA increased from 2.86% to 4.83% (p<0.05, n=8). The 95% detection limit of MCH was determined at 286 pg/ml (43 geg/ml) compared with 138 pg/ml (21 geq/ml) for the QIAamp DSP Virus Kit.

[Functional Genomic Approaches to Explore Secondary Metabolites in Medicinal Plants]

Zhongguo Zhong Yao Za Zhi = Zhongguo Zhongyao Zazhi = China Journal of Chinese Materia Medica. Jan, 2009  |  Pubmed ID: 19382439

Extensive genomic data concerning medicinal plants are rather scarce and insights of the secondary metabolic pathways and their regulatory mechanism are insufficient, hampering the broad application of cell or tissue cultivation and metabolic engineering to producing high-value secondary metabolites. The integration of cDNA-AFLP based transcript profiling and metabolomics, a new development of functional genomic approaches could establish correlations between the changes of secondary metabolites and expressions of related genes. It has manifested widely applicative prospects in seeking genes involved in biosynthesis of secondary metabolites and exploring secondary metabolic pathways. Functional genomic approaches are promising trends in the field of medicinal plants secondary metabolites research and will lead to better utilization of natural medicinal resources.

Measurement of the Membrane Elasticity of Red Blood Cell with Osmotic Pressure by Optical Tweezers

Cryo Letters. Mar-Apr, 2009  |  Pubmed ID: 19448857

Cells have to undergo many changes in osmotic pressure during their long-term preservation, and will have injuries before they return to their normal states. Mechanics of a cell with deformation, either small or large, is usually used to describe the change of the cell quantitatively. However, there are few reports on the deformation of cells subjected to the change of osmotic pressures during preservation. Here, we report our study of the elasticity of the human red blood cells under osmotic pressures using optical tweezers. We find that the deformation characteristics of erythrocytes are strongly dependent on the osmotic pressure. We also find the RBCs will become stiff with increasing osmotic pressure, suggesting a potential reason for membrane injury during preservation.

Results of Surgical Treatment of Cervical Dumbbell Tumors: Surgical Approach and Development of an Anatomic Classification System

Spine. May, 2009  |  Pubmed ID: 19455006

A retrospective study of a new classification and surgical approach of cervical dumbbell tumors.

The Effect of Relative Humidity on Binary Gas Diffusion

The Journal of Physical Chemistry. B. Jun, 2009  |  Pubmed ID: 19514781

The dependence of diffusion coefficient of O2-N2 mixture in the presence of water vapor was experimentally determined as a function of relative humidity (RH) with different temperatures using an in-house made Loschmidt diffusion cell. The experimental results showed that O2-N2 diffusion coefficient increased more than 17% when RH increased from 0% to 80% at 79 degrees C. In the experiments, the RH in both top and bottom chambers of the diffusion cell were the same, and the pressure inside the diffusion cell was kept as ambient pressure (1 atm.). Maxwell-Stefan theory was employed to analyze the mass transport in the diffusion cell, and found that there was no effective water vapor diffusion taking place, indicating that the gas diffusion in this ternary (O2-N2-water vapor) system could be considered binary gas (O2-N2) diffusion. The Fuller, Schettler, and Giddings (FSG) empirical equation of the kinetic theory of gases was generalized to accommodate the dependence of the binary diffusion coefficient on the RH. The prediction of the generalized equation was found to be consistent with experimental results with the difference of less than 1.5%, showing that the generalized equation could be applied to calculate the diffusion coefficients of the binary gaseous mixture with different temperature and RH values. The effect of water vapor on the increase of O2-N2 diffusion coefficient was discussed using molecule theory.

Ambulatory Arterial Stiffness Index is Not Correlated with the Pressor Response to Laboratory Stressors in Normotensive Humans

Journal of Hypertension. Apr, 2009  |  Pubmed ID: 19516175

Ambulatory arterial stiffness index (AASI) is a novel estimate of arterial stiffness, which independently predicts cardiovascular mortality, even in normotensive individuals. Additionally, other markers derived from ambulatory blood pressure (BP) monitoring, including variability, pulse pressure, nocturnal dipping, and morning BP surge, have all been shown to be predictive of end-organ damage and cardiovascular disease. Exaggerated cardiovascular reactivity to sympathoexcitatory stimuli may also predict future incidence of hypertension. The purpose of this investigation was to test the hypothesis that AASI and other derivations of ambulatory BP, including pulse pressure, 24-h blood pressure variability, dipping, and morning surge, would be correlated with the pressor response to common physiological stress maneuvers.

Birth of Parthenote Mice Directly from Parthenogenetic Embryonic Stem Cells

Stem Cells (Dayton, Ohio). Sep, 2009  |  Pubmed ID: 19544532

Mammalian parthenogenetic embryos are not viable and die because of defects in placental development and genomic imprinting. Parthenogenetic ESCs (pESCs) derived from parthenogenetic embryos might advance regenerative medicine by avoiding immuno-rejection. However, previous reports suggest that pESCs may fail to differentiate and contribute to some organs in chimeras, including muscle and pancreas, and it remains unclear whether pESCs themselves can form all tissue types in the body. We found that derivation of pESCs is more efficient than of ESCs derived from fertilized embryos, in association with reduced mitogen-activated protein kinase signaling in parthenogenetic embryos and their inner cell mass outgrowth. Furthermore, in vitro culture modifies the expression of imprinted genes in pESCs, and these cells, being functionally indistinguishable from fertilized embryo-derived ESCs, can contribute to all organs in chimeras. Even more surprisingly, our study shows that live parthenote pups were produced from pESCs through tetraploid embryo complementation, which contributes to placenta development. This is the first demonstration that pESCs are capable of full-term development and can differentiate into all cell types and functional organs in the body.

Iron (III) Citrate Inhibits Polyethylenimine-mediated Transient Transfection of Chinese Hamster Ovary Cells in Serum-free Medium

Cytotechnology. Jun, 2009  |  Pubmed ID: 19557539

Recent advances in transient transfection protocols using polyethylenimine (PEI) as a transfection reagent have led to the development of economical methods that provide yields sufficient for industrial production of proteins for many preclinical needs. There are many variables that can be optimized to improve protein expression in transient transfection, and one of the most critical is the medium in which the cells are grown. While transfection with PEI works well in media containing serum, the biopharmaceutical industry is moving away from animal-derived components in media. A number of serum-free media have been found to allow transient transfection, but many others do not for reasons that are not clear. Thus, knowledge of the components of serum-free media that can cause inhibition of PEI-mediated transient transfection would be useful for media development. In this study, an analysis was performed of various components of a serum-free medium used for Chinese hamster ovary cells in which PEI-mediated transient transfection was inhibited. We found that an iron supplement added to the medium was responsible for the inhibition. Further investigation showed that iron (III) citrate, a common iron chelator found in serum-free medium, was the specific component that caused the effect. Further, we showed that inhibition of transient transfection was caused by iron (III) citrate specifically, rather than citrate or iron alone. Finally, we showed that various iron chelators in serum-free media other than iron (III) citrate do not inhibit antibody expression.

Genetic Alterations in Differentiated Thyroid Cancers

Endocrine, Metabolic & Immune Disorders Drug Targets. Sep, 2009  |  Pubmed ID: 19594419

Differentiated thyroid cancers are the predominant malignancies of the thyroid. Due to advances in the understanding of the activation of the cell proliferation pathway at a molecular level, multiple genetic alterations have been linked to the development of thyroid carcinogenesis. Although the genetic alterations can be categorized into 7 categories, the BRAF mutation, RET/PTC, Pax8/PPARGamma, and dysfunctional Fas pathway have been most commonly described. Each of the gene alterations can ultimately result in cancer development, invasion and/or metastasis. This article provides a detailed review of the altered cell proliferation pathway activations found in thyroid carcinogenesis. The molecular targets that may be disrupted by therapeutic agents during the abnormal proliferation are also summarized.

Does the Onset of Sexual Maturation Terminate the Expression of Migratory Behaviour in Moths? A Study of the Oriental Armyworm, Mythimna Separata

Journal of Insect Physiology. Nov, 2009  |  Pubmed ID: 19643107

It is generally accepted that in most insects adults are sexually immature when they initiate migration and that migratory behaviour terminates with the onset of sexual maturation. However, a few studies examining the mating status of field collected moths have suggested that sexually mature individuals may continue migrating, but in these cases it was impossible to completely eliminate the possibility that the mated females captured came from local, non-migrant populations. In this study we examined the ovarian development of Mythimna separata females captured using a vertical pointing searchlight trap on Beihuang Island in the Bohai Gulf, China, a site >40km from land. Moths were collected from May to October from 2003 to 2008 in order to test the hypothesis that the onset of sexual maturation resulted in the termination of migratory behaviour. While females at the end of the summer had little ovarian development and were unmated, a significant proportion of those migrating northward in the early summer had developed ovaries and often had at least one spermatophore. Given that theses insects were captured while flying up to 500m above sea level, at a site with no local populations, the findings would not support the hypothesis and suggest that both ovarian development and mating may occur during migration.

Inhibition of Thioredoxin Reductase by Mansonone F Analogues: Implications for Anticancer Activity

Chemico-biological Interactions. Jan, 2009  |  Pubmed ID: 18822278

Mammalian thioredoxin reductase (TrxR), a ubiquitous selenocysteine containing oxidoreductase, catalyzes the NADPH-dependent reduction of oxidized thioredoxin (Trx). TrxR has been suggested as a potential target for anticancer drugs development for its overexpression in human tumors and its diverse functions in intracellular redox control, cell growth and apoptosis. Mansonone F (MF) compounds have been shown to exhibit antiproliferative effects, but their complex mechanisms are unknown. In the present study, we have investigated the effects of some synthesized MF analogues on TrxR and HeLa cells. The studies of the mode of inhibition and the interactions of IG3, one of the most potent MF analogues, with TrxR showed MF compounds could be partly reduced by the C-terminal selenolthiol active site, and possibly by the N-terminal dithiol motif and/or FAD domain of TrxR simultaneously, accompanied by redox cycling with the generation of superoxide anion radicals. In addition, MF analogues exhibited the potential to inhibit the growth of HeLa cells and reduce TrxR activity in cell lysates. The cell cycle was arrested in G2/M phase and apoptosis was induced in a dose-dependent manner. Furthermore, our results showed that IG3-treated HeLa cells induced the change of intracellular ROS. Taken together, the reported results here suggest that inhibition of TrxR by MF analogues provides a possible complex mechanism for explaining the anticancer activity of MF compounds.

[Surveys on Resources and Varieties on Chinese Markets of Crude Drug Xixin]

Zhongguo Zhong Yao Za Zhi = Zhongguo Zhongyao Zazhi = China Journal of Chinese Materia Medica. Dec, 2010  |  Pubmed ID: 21438380

Many species of the genus Asarum plants are used as Chinese traditional or folk medicines, in Chinese which are known as "Xixin" (Asari Radix et Rhizoma) , "Bei-Xixin" , "Hua-Xixin" and "Nan-Xixin" , etc. In order to get a clear picture of resources distribution and varieties on Chinese markets of the crude drug Xixin and provide scientific basis for their resource conservation and sustainable use, during recent years we conducted field investigations and market researches many times. The results showed that the resources of both official Xixins and non-official Xixins were decreased because of the ecological environment damage and over-digging, especially species whose population size was small. Bei-Xixin derived from A. heterotropoides var. mandshuricum was the most species on the Chinese markets except for a few areas of China. Hua-Xixin derived from A. sieboldii and non-official Xixins were mainly used in their producing areas. Cultivation of Hua-Xixin should be greatly developed, and wild resources of non-official species must be preserved strictly.

Langerhans Cell Histiocytosis of the Cervical Spine: a Single Chinese Institution Experience with Thirty Cases

Spine. Jan, 2010  |  Pubmed ID: 20042947

A retrospective study of cervical Langerhans cell histiocytosis (LCH).

[Breeding and Identification of Estrogen Receptor Beta Gene Knock-out Mice]

Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University. Jan, 2010  |  Pubmed ID: 20118009

To breed estrogen receptor beta (ERbeta) gene knock-out female mice for studying postmenopausal osteoporotic fracture.

MRNA Stability and Antibody Production in CHO Cells: Improvement Through Gene Optimization

Biotechnology Journal. Apr, 2010  |  Pubmed ID: 20222103

The productivity of stably transfected cell lines is of critical importance for the manufacturing of therapeutic proteins. Various methods have been successfully implemented to increase the production output of mammalian cell cultures. Increasing evidence suggests that optimization of the gene coding sequences of an expression vector can improve specific cell line yield of the recombinant protein. Here we demonstrate that gene optimization substantially enhances antibody production in Chinese hamster ovary cells. When gene optimization was applied to the heavy and light chain genes of a therapeutic antibody, we observed increased antibody production in transient transfection. Elevated heavy chain mRNA level was associated with the increase of antibody production. Further analysis suggested that the increased antibody expression is attributable to enhanced mRNA stability resulting from gene optimization. Gene optimization also led to increased antibody production in stable clones.

Metabolic Syndrome and All-cause Mortality: a Meta-analysis of Prospective Cohort Studies

European Journal of Epidemiology. Jun, 2010  |  Pubmed ID: 20425137

To synthesize the available data on the association between metabolic syndrome and all-cause mortality, we conducted a meta-analysis of prospective cohort studies. We performed a literature search using Medline, EMBASE and Cochrane Library from 2001 to December 2009, with no restrictions. We included studies if they were prospective, had an assessment of metabolic syndrome at baseline and risk of all-cause mortality. We recorded several characteristics for each study. We extracted relative risks (RR) and 95% confidence intervals (CI) and pooled them using fixed or random effects models. We performed sensitivity analysis, and assessed heterogeneity and publication bias. A total of 21 studies including 372,411 participants were included in our meta-analysis. 18,556 deaths from any cause occurred during a mean follow-up of 11.5 years. Individuals with the metabolic syndrome, compared to those without, had an increased mortality from all causes (pooled RR 1.46; 95% CI 1.35-1.57). The RR of all-cause mortality associated with metabolic syndrome was higher in studies using the National Cholesterol Education Program Adult Treatment Panel (NCEP) than the revised NCEP criteria (RR: 1.45 vs. 1.25; P = 0.0002). Metabolic syndrome is an important risk factor for all-cause mortality. The diagnosis and treatment of the underlying risk factors for the metabolic syndrome should be an important strategy for the reduction of all-cause mortality associated with metabolic syndrome in the general population.

Langerhans Cell Histiocytosis of the Atlas in an Adult

European Spine Journal : Official Publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society. Jan, 2010  |  Pubmed ID: 19844749

Langerhans cell histiocytosis (LCH), formerly known as histiocytosis X, is a rare disorder (approximately 1:1,500,000 inhabitants) characterized by clonal proliferation and excess accumulation of pathologic Langerhans cells causing local or systemic effects. The exact etiology of LCH is still unknown. LCH could affect patients of any age, although most present when they are children. The most frequent sites of the bony lesions are the skull, femur, mandible, pelvis and spine. A variety of treatment modalities has been reported, but there was no evidence suggesting that any one treatment was more advantageous than another. We present an adult with LCH of the atlas. A 26-year-old young man presented with a 2-month history of neck pain and stiffness. CT revealed osteolytic lesion in the left lateral mass of atlas with compression fracture. Histopathological diagnosis was Langerhans cell histiocytosis by percutaneous needle biopsy under CT guidance. The patient underwent conservative treatment, including Halo-vest immobilization and radiotherapy. At 7-year follow-up, the patient was asymptomatic except for mild motion restriction of the neck. CT revealed a significant reconstruction of the C1 lateral mass.

A Noncoding Antisense RNA in Tie-1 Locus Regulates Tie-1 Function in Vivo

Blood. Jan, 2010  |  Pubmed ID: 19880500

Recently, messenger RNAs in eukaryotes have shown to associate with antisense (AS) transcript partners that are often referred to as long noncoding RNAs (lncRNAs) whose function is largely unknown. Here, we have identified a natural AS transcript for tyrosine kinase containing immunoglobulin and epidermal growth factor homology domain-1 (tie-1), tie-1AS lncRNA in zebrafish, mouse, and humans. In embryonic zebrafish, tie-1AS lncRNA transcript is expressed temporally and spatially in vivo with its native target, the tie-1 coding transcript and in additional locations (ear and brain). The tie-1AS lncRNA selectively binds tie-1 mRNA in vivo and regulates tie-1 transcript levels, resulting in specific defects in endothelial cell contact junctions in vivo and in vitro. The ratio of tie-1 versus tie-1AS lncRNA is altered in human vascular anomaly samples. These results directly implicate noncoding RNA-mediated transcriptional regulation of gene expression as a fundamental control mechanism for physiologic processes, such as vascular development.

Prevention of Graft-versus-host Diseases by in Vivo SupCD28mAb-expanded Antigen-specific NTreg Cells

Cell Transplantation. 2010  |  Pubmed ID: 20573297

Naturally occurring CD4(+)CD25(+) Treg cells (nTregs) can be exploited to establish an immunologic tolerance to non-self-antigens. The in vivo administration of a single superagonistic CD28-specific monoclonal antibody (supCD28mAb) to naive rat preferentially expanded the nTregs, which induced a potent inhibition of lethality of the graft-versus-host (GvH) diseases. The appearance of increased Foxp3 molecules was accompanied with a polarization towards a Th2 cytokine profile with a decreased production of IFN-γ and increased production of IL-4 and IL-10 in the serum of the antibody-treated rat. The peripheral Foxp3 nTregs are decreased in acute GvHD, while supCD28mAb administration showed that nTregs were preferentially proliferating in vivo, thus resulting in the significant prevention of the GvH disease. Furthermore, antigen-specific nTregs could suppress conventional T-cell proliferation stimulated with alloantigen in vitro. Taken together, our findings demonstrate that the potent regulatory functions of the Tregs for the treatment of GvHD are antigen specific. These data also provide evidence that GvHD is associated with decrease of Tregs in the periphery of the host. The determination of the Foxp3 Tregs can be a helpful tool to discriminate GvHD severity and lethality after allogeneic stem cell transplantation.

Effects of Vitamin D Supplementation on Blood Pressure

Southern Medical Journal. Aug, 2010  |  Pubmed ID: 20622727

Inconsistent findings from epidemiological studies have continued the controversy over the role of oral vitamin D supplementation in reducing blood pressure in normotensive or hypertensive populations.

Determination of SNX-2112, a Selective Hsp90 Inhibitor, in Plasma Samples by High-performance Liquid Chromatography and Its Application to Pharmacokinetics in Rats

Journal of Pharmaceutical and Biomedical Analysis. Dec, 2010  |  Pubmed ID: 20675090

A sensitive and specific reversed-phase high-performance liquid chromatography method with ultraviolet detection has been developed and validated for the identification and quantification of SNX-2112 in rat plasma. Following sample preparation using liquid-liquid extraction, the analytes were separated by the mobile phase acetonitrile-water (40:60, v/v) with an Agilent RP-HPLC column (ZORBAX SB-C18, 5 microm, 4.6 mm x 250 mm) at a flow rate of 1 ml/min, column temperature of 30 degrees C and detection wavelength of 251 nm. The retention time of SNX-2112 was 11.2 min. A good linear relationship was obtained in the concentration range studied (0.07-21 microg/ml, R(2)>0.9982), and the LLOD and LLOQ for SNX-2112 were 0.02 and 0.07 microg/ml, respectively. The mean absolute recovery of SNX-2112 in plasma ranged from 88.58 to 99.61% at the studied concentrations. The intra- and inter-batch relative standard deviations were 1.7-3.5 and 1.9-4.4%, respectively. This method was successfully applied to pharmacokinetic studies in rats after intravenous administration of SNX-2112.

Bufogargarizins A and B: Two Novel 19-norbufadienolides with Unprecedented Skeletons from the Venom of Bufo Bufo Gargarizans

Chemistry (Weinheim an Der Bergstrasse, Germany). Sep, 2010  |  Pubmed ID: 20715201

Nogo-B Receptor is Essential for Angiogenesis in Zebrafish Via Akt Pathway

Blood. Dec, 2010  |  Pubmed ID: 20813898

Our previous work has shown that axon guidance gene family Nogo-B and its receptor (NgBR) are essential for chemotaxis and morphogenesis of endothelial cells in vitro. To investigate NogoB-NgBR function in vivo, we cloned the zebrafish ortholog of both genes and studied loss of function in vivo using morpholino antisense technology. Zebrafish ortholog of Nogo-B is expressed in somite while expression of zebrafish NgBR is localized in intersomitic vessel (ISV) and axial dorsal aorta during embryonic development. NgBR or Nogo-B knockdown embryos show defects in ISV sprouting in the zebrafish trunk. Mechanistically, we found that NgBR knockdown not only abolished its ligand Nogo-B-stimulated endothelial cell migration but also reduced the vascular endothelial growth factor (VEGF)-stimulated phosphorylation of Akt and vascular endothelial growth factor-induced chemotaxis and morphogenesis of human umbilical vein endothelial cells. Further, constitutively activated Akt (myristoylated [myr]Akt) or human NgBR can rescue the NgBR knockdown umbilical vein endothelial cell migration defects in vitro or NgBR morpholino-caused ISV defects in vivo. These data place Akt at the downstream of NgBR in both Nogo-B- and VEGF-coordinated sprouting of ISVs. In summary, this study identifies the in vivo functional role for Nogo-B and its receptor (NgBR) in angiogenesis in zebrafish.

Structural and Degradation Characteristics of an Innovative Porous PLGA/TCP Scaffold Incorporated with Bioactive Molecular Icaritin

Biomedical Materials (Bristol, England). Oct, 2010  |  Pubmed ID: 20876954

Phytomolecules may chemically bind to scaffold materials for medical applications. The present study used an osteoconductive porous poly(l-lactide-co-glycolide)/tricalcium phosphate (PLGA/TCP) to incorporate an exogenous phytoestrogenic molecule icaritin to form a PLGA/TCP/icaritin composite scaffold material with potential slow release of icaritin during scaffold degradation. Accordingly, the present study was designed to investigate its in vitro degradation characteristics and the release pattern of icaritin at three different doses (74 mg, 7.4 mg and 0.74 mg per 100 g PLGA/TCP, i.e. in the PLGA/TCP/icaritin-H, -M and -L groups, respectively). A PLGA/TCP/icaritin porous composite scaffold was fabricated using a computer-controlled printing machine. The PLGA/TCP/icaritin scaffolds were incubated in saline at 37 °C for 12 weeks and the pure PLGA/TCP scaffold served as a control. During the 12 weeks in vitro degradation, the scaffolds in all four groups showed changes, including a decrease in weight, volume and pore size of the composite scaffold, while there was a decrease in acidity and an increase in Ca and lactic acid concentrations in the degradation medium, especially after 7 weeks. The rate of degradation was explained by the relationship with the content of icaritin incorporated into the scaffolds. The higher the icaritin content in the scaffolds, the slower the degradation could be observed during 12 weeks. After 12 weeks, the SEM showed that the surface of the PLGA/TCP and PLGA/TCP/icaritin-L groups was relatively smooth with a gradual decrease in number and size of the micropores, while the porous morphology on the surface of the PLGA/TCP/icaritin-M and PLGA/TCP/icaritin-H groups was partly maintained, accompanied by a decrease in phosphate (P) and calcium (Ca) contents at the surface. Though the mechanical property of the PLGA/TCP/icaritin scaffold decreased after degradation, its porous structure was maintained, which was essential for cell migration and ingrowth of newly regenerated tissues in vivo. The controlled release of icaritin from the composite scaffold reached about 70% of the incorporated icaritin into the degradation medium after 12 weeks. The above findings suggested that the structural and degradation properties of the porous composite PLGA/TCP/icaritin scaffold were dependent on icaritin concentrations. This innovative composite porous scaffold material developed in the present study may be used as a good scaffold material for enhancing bone repair, especially at high concentrations of icaritin. In vivo confirmation is, however, needed to substantiate our in vitro findings.

Psiguadials A and B, Two Novel Meroterpenoids with Unusual Skeletons from the Leaves of Psidium Guajava

Organic Letters. Nov, 2010  |  Pubmed ID: 20929258

Psiguadials A (1) and B (2), two novel sesquiterpenoid-diphenylmethane meroterpenoids with unusual skeletons, along with a pair of known epimers, psidial A (3) and guajadial (4), were isolated from the leaves of Psidium guajava. Their structures with absolute configurations were elucidated by means of NMR, X-ray diffraction, and quantum chemical CD calculation. Compounds 1, 2, and 4 exhibited potent inhibitory effects on the growth of human hepatoma cells.

Antenatal Rh Prophylaxis is Unnecessary for "Asia Type" DEL Women

Transfusion Clinique Et Biologique : Journal De La Société Française De Transfusion Sanguine. Oct, 2010  |  Pubmed ID: 20961786

Generally Rh-negative patients need to be transfused with Rh-negative red blood cells. For pregnant women carrying Rh-positive fetus, the antenatal anti-D detection and Rh immunoglobulin prophylaxis are required worldwide. In East Asia, a RhD variant, termed "Asia type" DEL, was found in approximately 30% of apparent Rh-negative individuals. The antigenic and molecular properties of the DEL were previously defined. Few data discuss whether DEL could be immunized by D antigen clinically although DEL was reported arousing alloimmunization to true Rh-negative patients.

Simvastatin Prevents ERK Activation in Myocardial Hypertrophy of Spontaneously Hypertensive Rats

Scandinavian Cardiovascular Journal : SCJ. Dec, 2010  |  Pubmed ID: 21080865

Statins exert regression of left ventricular hypertrophy independent of their plasma cholesterol-lowering actions. However, the underlying mechanism is not clear.

Highly Selective Detection of Phosphate in Very Complicated Matrixes with an Off-on Fluorescent Probe of Europium-adjusted Carbon Dots

Chemical Communications (Cambridge, England). Mar, 2011  |  Pubmed ID: 21234476

A simple method for phosphate (Pi) detection is established by developing an off-on fluorescence probe of europium-adjusted carbon dots (CDs), which has been successfully applied to the detection of Pi in very complicated matrixes such as artificial wetlands system.

Germline Competency of Parthenogenetic Embryonic Stem Cells from Immature Oocytes of Adult Mouse Ovary

Human Molecular Genetics. Apr, 2011  |  Pubmed ID: 21239471

Parthenogenetic embryonic stem cells (pESCs) have been generated in several mammalian species from parthenogenetic embryos that would otherwise die around mid-gestation. However, previous reports suggest that pESCs derived from in vivo ovulated (IVO) mature oocytes show limited pluripotency, as evidenced by low chimera production, high tissue preference and especially deficiency in germline competence, a critical test for genetic integrity and pluripotency of ESCs. Here, we report efficient generation of germline-competent pESC lines (named as IVM pESCs) from parthenogenetic embryos developed from immature oocytes of adult mouse ovaries following in vitro maturation (IVM) and artificial activation. In contrast, pESCs derived from IVO oocytes show defective germline competence, consistent with previous reports. Further, IVM pESCs resemble more ESCs from fertilized embryos (fESCs) than do IVO pESCs on genome-wide DNA methylation and global protein profiles. In addition, IVM pESCs express higher levels of Blimp1, Lin28 and Stella, relative to fESCs, and in their embryoid bodies following differentiation. This may indicate differences in differentiation potentially to the germline. The mechanisms for acquisition of pluripotency and germline competency of IVM pESCs from immature oocytes remain to be determined.

Binuclear Methylborole Iron Carbonyls: Iron-iron Multiple Bonds and Perpendicular Structures

Inorganic Chemistry. Feb, 2011  |  Pubmed ID: 21250640

Methylborole iron tricarbonyl, (η(5)-C(4)H(4)BCH(3))Fe(CO)(3), is known experimentally and is a potential source of binuclear (C(4)H(4)BCH(3))(2)Fe(2)(CO)(n) (n = 5, 4, 3, 2, 1) derivatives through reactions such as photolysis. In this connection the lowest energy (C(4)H(4)BCH(3))(2)Fe(2)(CO)(5) structures are predicted theoretically to have a single bridging carbonyl group and Fe-Fe distances consistent with formal single bonds. The lowest energy (C(4)H(4)BCH(3))(2)Fe(2)(CO)(4) structures have two bridging carbonyl groups and Fe═Fe distances suggesting formal double bonds. Analogously, the lowest energy (C(4)H(4)BCH(3))(2)Fe(2)(CO)(3) structures have three bridging carbonyl groups and very short Fe≡Fe distances suggesting formal triple bonds. The tetracarbonyl (C(4)H(4)BCH(3))(2)Fe(2)(CO)(4) is predicted to be thermodynamically unstable toward disproportionation into (C(4)H(4)BCH(3))(2)Fe(2)(CO)(5) + (C(4)H(4)BCH(3))(2)Fe(2)(CO)(3), whereas the tricarbonyl is thermodynamically viable toward analogous disproportionation. The lowest energy structures of the more highly unsaturated methylborole iron carbonyls (C(4)H(4)BCH(3))(2)Fe(2)(CO)(n) (n = 2, 1) have hydrogen atoms bridging an iron-carbon bond. In addition, the lowest energy (C(4)H(4)BCH(3))(2)Fe(2)(CO) structures are "slipped perpendicular" structures with bridging methylborole ligands, a terminal carbonyl group, and agostic CH(3)→Fe interactions involving the methyl hydrogens. Thus, in these highly unsaturated systems the methyl substituent in the methylborole ligand chosen in this work is not an "innocent bystander" but instead participates in the metal-ligand bonding.

Langerhans Cell Histiocytosis with Multiple Spinal Involvement

European Spine Journal : Official Publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society. Nov, 2011  |  Pubmed ID: 20496040

To stress the clinical and radiologic presentation and treatment outcome of Langerhans cell histiocytosis (LCH) with multiple spinal involvements. A total of 42 cases with spinal LCH were reviewed in our hospital and 5 had multifocal spinal lesions. Multiple spinal LCH has been reported in 50 cases in the literature. All cases including ours were analyzed concerning age, sex, clinical and radiologic presentation, therapy and outcome. Of our five cases, three had neurological symptom, four soft tissue involvement and three had posterior arch extension. Compiling data from the eight largest case series of the spinal LCH reveals that 27.2% multiple vertebrae lesions. In these 55 cases, there were 26 female and 29 male with the mean age of 7.4 years (range 0.2-37). A total of 182 vertebrae were involved including 28.0% in the cervical spine, 47.8% in thoracic and 24.2% in the lumbar spine. Extraspinal LCH lesion was documented in 54.2% cases, visceral involvement in 31.1% and vertebra plana in 50% cases. Paravertebral and epidural extension were not documented in most cases. Pathological diagnosis was achieved in 47 cases including 8 open spine biopsy. The treatment strategy varied depending on different hospitals. One patient died, two had recurrence and the others had no evidence of the disease with an average of 7.2 years (range 1-21) of follow-up. Asymptomatic spinal lesions could be simply observed with or without bracing and chemotherapy is justified for multiple lesions. Surgical decompression should be reserved for the uncommon cases in which neurologic compromise does not respond to radiotherapy or progresses too rapidly for radiotherapy.

A Dilution-filtration System for Removing Cryoprotective Agents

Journal of Biomechanical Engineering. Feb, 2011  |  Pubmed ID: 21280879

In most cryopreservation applications, the final concentrations of cryoprotective agents (CPAs) must be reduced to biocompatible levels. However, traditional methods for removing CPAs usually have disadvantages of operation complexity, time consumption, and ease of contamination, especially for the applications involving large volumes of cell suspensions. A dilution-filtration system, which involves pure ultrafiltration for separation, was developed for continuous, automatic, and closed process of removing CPAs. To predict the optimal protocols under given experimental conditions, a theoretical model was established first. Cell-free experiments were then conducted to investigate the variation in CPA concentration during the process, and the experimental data were compared with the theoretical values for the validation of the model. Finally, ten units (212.9 ml/unit±9.5 ml/unit) of thawed human red blood cells (cryopreserved with 40% (w/v) glycerol) were deglycerolized using the theoretically optimal operation protocols to further validate the effectiveness and advantage of the system. In the cell-free experiments, glycerol was continuously removed and the concentration variations fitted the simulated results quite well. In the in-vitro experiments, glycerol concentration in RBC suspension was reduced to 5.57 g/l±2.81 g/l within an hour, and the cell count recovery rate was 91.19%±3.57%, (n=10), which proves that the system is not only safe for removing CPAs, but also particularly efficient for processing large-scale samples. However, the operation parameters must be carefully controlled and the optimal protocols should be specialized and various from case to case. The presented theoretical model provides an effective approach to find out the optimal operation protocols under given experimental conditions and constrains.

Induction of Apoptosis in K562 Cells by Dicyclohexylammonium Salt of Hyperforin Through a Mitochondrial-related Pathway

Chemico-biological Interactions. Apr, 2011  |  Pubmed ID: 21376709

Hyperforin is an abundant phloroglucinol-type constituent isolated from the extract of the flowering upper portion of the plant Hypericum perforatum L. The dicyclohexylammonium salt of hyperforin (DCHA-HF) has exhibited antitumor and antiangiogenic activities in various cancer cells. Here, the antitumor effects of DCHA-HF on the chronic myeloid leukemia K562 cell line were investigated for the first time. DCHA-HF exhibited dose- and time-dependent inhibitory activities against K562 cells, with IC(50) values of 8.6 and 3.2 μM for 48 h and 72 h of treatment, respectively, which was more effective than that of the hyperforin. In contrast, little cytotoxic activity was observed with DCHA-HF on HUVECs. DCHA-HF treatment resulted in induction of apoptosis as evidenced from DNA fragmentation, nuclear condensation and increase of early apoptotic cells by DAPI staining analysis, TUNEL assay and Annexin V-FITC/PI double-labeled staining analysis, respectively. Moreover, DCHA-HF elicited dissipation of mitochondrial transmembrane potential that commenced with the release of cytochrome c through down-regulation of expression of anti-apoptotic proteins and up-regulation of expression of pro-apoptotic proteins. DCHA-HF treatment induced activation of the caspase 3, 8, and 9 cascade and subsequent PARP cleavage, and DCHA-HF-induced apoptosis was significantly inhibited by caspase inhibitors. Treated cells were arrested at the G1 phase of the cell cycle and the expression of p53 and p27(Kip1), two key regulators related to cell cycle and apoptosis, was up-regulated. These results suggest that DCHA-HF inhibits K562 cell growth by inducing caspase-dependent apoptosis mediated by a mitochondrial pathway and arresting the cell cycle at the G1 phase. Therefore, DCHA-HF is a potential chemotherapeutic antitumor drug for chronic myeloid leukemia therapy.

Impaired Bone Healing Pattern in Mice with Ovariectomy-induced Osteoporosis: A Drill-hole Defect Model

Bone. Jun, 2011  |  Pubmed ID: 21421090

To establish a drill-hole defect model in osteoporotic mouse femur by comparing temporal cortical bone healing pattern between OVX-induced osteoporotic bone and sham-operated bone.

[Effects of Garlic Oil, Age and Sex on N-hexane Metabolism in Rats]

Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi = Zhonghua Laodong Weisheng Zhiyebing Zazhi = Chinese Journal of Industrial Hygiene and Occupational Diseases. Jan, 2011  |  Pubmed ID: 21619798

To investigate effects of garlic oil (GO), age and sex on n-hexane metabolism in rats.

BJ-B11, a Novel Hsp90 Inhibitor, Induces Apoptosis in Human Chronic Myeloid Leukemia K562 Cells Through the Mitochondria-dependent Pathway

European Journal of Pharmacology. Sep, 2011  |  Pubmed ID: 21620825

In the past few years heat shock protein 90 (Hsp90) inhibitors have been reported to possess significant antitumor activity. We investigated, for the first time, the antitumor activity of a novel Hsp90 inhibitor 2-(4-acetyloxycyclohexylamino)-4-(3, 6, 6-trimethyl-4-oxo-4, 5, 6, 7-tetrahydro-1H-indazol-1-yl)-benzamide (BJ-B11) and the molecular mechanism underlying the apoptosis it induces in human chronic myeloid leukemia K562 cells. The results revealed that BJ-B11 triggered growth inhibition in K562 cells and other malignant cell lines in vitro with only minor toxicity in a normal human cell line. BJ-B11 inhibited the proliferation of K562 cells in a concentration- and time-dependent manner, with IC(50) values of 1.1 ± 0.2 μM and 0.4 ± 0.1 μM after 48 and 72 h incubations respectively. This most likely results from cell cycle arrest at the G(0)/G(1) phase and the induction of apoptosis. In addition, BJ-B11 degraded the Hsp90 client proteins Bcr-Abl and Akt, induced activation of caspase-9 and caspase-3, and subsequent cleavage of PARP. The caspase signals may originate from mitochondrial dysfunction, which is supported by the finding of cytochrome c release. In addition, inactivation of the Akt signaling pathway may be involved in the process of BJ-B11-induced apoptosis. Taken together, our data provide a putative molecular mechanism for the anticancer effect of BJ-B11 on K562 cells, and suggest a potential application for BJ-B11 in chronic myeloid leukemia therapy.

Sulfation of Selected Mono-hydroxyflavones by Sulfotransferases in Vitro: a Species and Gender Comparison

The Journal of Pharmacy and Pharmacology. Jul, 2011  |  Pubmed ID: 21635263

Sulfation via sulfotransferases is an important metabolic pathway contributing to the low bioavailability of flavonoids. This study aims to characterize the sulfation of mono-hydroxyflavones (MHFs) to obtain useful information on structure-metabolizing relationships in animal species and gender differences.

[Advances in the Molecular Pathogenesis of Hypertrophic Cardiomyopathy]

Yi Chuan = Hereditas / Zhongguo Yi Chuan Xue Hui Bian Ji. Jun, 2011  |  Pubmed ID: 21684859

Hypertrophic Cardiomyopathy (HCM) is a primary cardiac disorder characterized by asymmetric thickening of the septum and left ventricular wall. HCM affects 1 in 500 individuals in the general population, and it is the most common cause of sudden death in the young and athletes. The clinic phenotype of HCM is highly variable with respect to age at onset, degree of symptoms, and risk of sudden death. HCM is usually inherited as a Mendelian autosomal dominant trait. To date, over 900 mutations have been reported in HCM, which were mainly located in 13 genes encoding cardiac sarcomere protein, e.g., MYH7, MYBPC3, and TnT. In addition, more and more mitochondrial DNA mutations were reported to be associated with the pathogenesis of HCM. Based on the description of the clinical phenotype and morphological characteristics, this review focuses on the research in the molecular pathogenic mechanism of HCM and its recent advances.

In-vitro Antitumor Activity Evaluation of Hyperforin Derivatives

Journal of Asian Natural Products Research. Aug, 2011  |  Pubmed ID: 21751836

The derivatives of hyperforin, namely hyperforin acetate (2), 17,18,22,23,27,28,32,33-octahydrohyperforin acetate (3), and N,N-dicyclohexylamine salt of hyperforin (4), have been investigated for their antitumor properties. In-vitro studies demonstrated that 2 and 4 were active against HeLa (human cervical cancer), A375 (human malignant melanoma), HepG2 (human hepatocellular carcinoma), MCF-7 (human breast cancer), A549 (human nonsmall cell lung cancer), K562 (human chronic myeloid leukemia), and K562/ADR (human adriamycin-resistant K562) cell lines with IC(50) values in the range of 3.2-64.1 μM. The energy differences between highest occupied molecular orbital and lowest unoccupied molecular orbital of 2-4 were calculated to be 0.39778, 0.43106, and 0.30900 a.u., respectively, using the Gaussian 03 software package and ab initio method with the HF/6-311 G* basis set. The result indicated that the biological activity of 4 might be the strongest and that of 3 might be the weakest, which was in accordance with their corresponding antiproliferative effects against the tested tumor cell lines. Compound 4 caused cell cycle arrest at G2/M phase in flow cytometry experiment and induced apoptosis by 4',6-diamidino-2-phenylindole staining and Annexin V-FITC/PI (propidium iodide) double-labeled staining in HepG2 cells. The results indicated a potential for N,N-dicyclohexylamine salt of hyperforin as a new antitumor drug.

SNX-2112, a Novel Hsp90 Inhibitor, Induces G2/M Cell Cycle Arrest and Apoptosis in MCF-7 Cells

Bioscience, Biotechnology, and Biochemistry. 2011  |  Pubmed ID: 21821931

SNX-2112 is a heat shock protein 90 (Hsp90) inhibitor with anticancer properties currently in clinical trials. This study investigated the effects of SNX-2112 on inhibition of cell growth, the cell cycle, and apoptosis in MCF-7 human breast cancer cells, in addition to the various molecular mechanisms. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometric analysis suggest that SNX-2112 inhibits cell growth in a time- and dose-dependent manner more potently than 17-(allylamino)-17-demethoxygeldanmycin (17-AAG), a traditional Hsp90 inhibitor, probably as a result of cell-cycle arrest at the G2/M phase and the induction of apoptosis. Downregulation of Bcl-2 and Bcl-xL, upregulation of Bax, cleavage of caspase-9 and poly (ADP-ribose) polymerase (PARP), and degradation of the breast cancer-related Hsp90 client proteins human epidermal growth factor receptor-2 (HER2), Akt, Raf-1, and nuclear factor kappa-B kinase (IKK) were observed in SNX-2112 treated cells by Western blot assay. These findings suggest that the molecular mechanisms of cell-growth inhibition by SNX-2112 involve activation of the mitochondrial apoptotic pathway and the degradation of breast cancer-related proteins.

Quantitative Proteomics Analysis of Parthenogenetically Induced Pluripotent Stem Cells

Protein & Cell. Aug, 2011  |  Pubmed ID: 21904979

Parthenogenetic embryonic stem (pES) cells isolated from parthenogenetic activation of oocytes and embryos, also called parthenogenetically induced pluripotent stem cells, exhibit pluripotency evidenced by both in vitro and in vivo differentiation potential. Differential proteomic analysis was performed using differential in-gel electrophoresis and isotope-coded affinity tag-based quantitative proteomics to investigate the molecular mechanisms underlying the developmental pluripotency of pES cells and to compare the protein expression of pES cells generated from either the in vivo-matured ovulated (IVO) oocytes or from the in vitro-matured (IVM) oocytes with that of fertilized embryonic stem (fES) cells derived from fertilized embryos. A total of 76 proteins were upregulated and 16 proteins were downregulated in the IVM pES cells, whereas 91 proteins were upregulated and 9 were downregulated in the IVO pES cells based on a minimal 1.5-fold change as the cutoff value. No distinct pathways were found in the differentially expressed proteins except for those involved in metabolism and physiological processes. Notably, no differences were found in the protein expression of imprinted genes between the pES and fES cells, suggesting that genomic imprinting can be corrected in the pES cells at least at the early passages. The germline competent IVM pES cells may be applicable for germ cell renewal in aging ovaries if oocytes are retrieved at a younger age.

Paeoniflorin Reduced Acute Toxicity of Aconitine in Rats is Associated with the Pharmacokinetic Alteration of Aconitine

Journal of Ethnopharmacology. Sep, 2011  |  Pubmed ID: 21930193

ETHNOPHARMACOLOGICAL RELEVANCE: To investigate the influence of paeoniflorin (major bioactive component of Paeonia lactiflora Pall.) on the pharmacokinetic behavior of aconitine (major toxic and bioactive component of Aconitum carmichaeli Debx.) and potential detoxifying effect of paeoniflorin on the acute toxicity of aconitine, which may provide in depth understanding to the toxicity reduction effect of Paeonia lactiflora to Aconitum carmichaeli. MATERIALS AND METHODS: Ultra high performance liquid chromatography coupled with triple quadrupole mass spectrometer (UHPLC-MS/MS) was employed to determine the plasma content of aconitine. Aconitine was administrated by oral to SD rats at the dosage of 200μg/kg with or without paeoniflorin given by intraperitoneal injection at the dosage of 20mg/kg. Plasma samples were collected for determination and analysis of pharmacokinetic parameters of aconitine. The LD(50) of aconitine and acute animal death induced by aconitine were examined when aconitine was given alone or jointly with paeoniflorin in ICR mice. RESULTS: A sensitive, accurate, precise, reliable and repeatable UHPLC-MS/MS method was successfully established for determination of the plasma content of aconitine in 12.5μL plasma sample. The lower limit of quantification of aconitine was 0.01ng/mL. Compared with the SD rats that were orally administrated with aconitine alone, the rats received aconitine and co-administrated with paeoniflorin by peritoneal injection showed a remarkably lower C(max) (5.69ng/mL vs 9.66ng/mL, P<0.05) and delayed T(max) (70min vs 46min, P<0.05), as well as a trend of reduction in AUC(0-t) (1082.75ng-min/mL vs 1650.27ng-min/mL, P=0.395). The LD(50) values of aconitine coadministered with 120 or 240mg/kg of paeoniflorin were obviously increased to 2.30 and 2.15mg/kg against 1.80mg/kg of aconitine by oral administration alone. Mice treated with paeoniflorin (240mg/kg) and aconitine (1.8mg/kg) together revealed a significant decreased death rate than that received aconitine treatment alone (15% vs 50%, P<0.05). CONCLUSIONS: The acute oral toxicity of aconitine in rats was significantly reduced by paeoniflorin; this might result from the alterations of pharmacokinetic behavior of aconitine in the animals by coadministration of paeoniflorin.

Carbon Nanotube-DNA Hybrid Used for Activity Monitoring and Inhibitor Screening of Nuclease

Analytica Chimica Acta. Nov, 2011  |  Pubmed ID: 21995925

Carbon nanotubes (CNTs) can efficiently quench the fluorescence of the adsorbed fluorophores and nonconvalently interact with soft single-stranded DNA (ssDNA). Upon disruption of CNTs-fluorescent oligonucleotides hybrid by nuclease S1, fluorescence turn-on was observed. Using this strategy, a platform based on fluorescence signal for monitoring the activity of nuclease with advantages of high sensitivity and commonality was established, and a linear relationship between initial cleavage reaction rate and nuclease S1 concentration is found in the range of 0.6-8.0 U mL(-1) with a detection limit of 0.08 U mL(-1). Furthermore, by taking pyrophosphate as an example, we use the assay to evaluate the prohibition effect on nuclease, and the extent of fluorescence recovery decreased linearly with increasing the concentration of pyrophosphate in the range of 0.2-1.4 mM, implying that the cleavage reaction by nuclease S1 was prohibited, and therefore this fluorescence assay can also be conveniently utilized for inhibitor screening of nuclease.

[Aeroallergen Spectrum of Patients with Allergic Rhinitis in Enshi Area]

Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi = Journal of Clinical Otorhinolaryngology, Head, and Neck Surgery. Jul, 2011  |  Pubmed ID: 22032124

To investigate aeroallergen spectrum of patients with allergic rhinitis in Enshi area and analyze its related factors.

Promoted Repair of Steroid-associated Osteonecrosis by Implantation of Cryopreserved Bone Marrow Mononuclear Cells in a Rabbit Model

Arthritis and Rheumatism. Nov, 2011  |  Pubmed ID: 22076746

OBJECTIVE: The cytotherapy can hardly promote repair in steroid-associated osteonecrosis (SAON). We found this was caused by impaired bioactivity of bone marrow stem cells (BMSCs) after pulsed administration of steroid. As the cryopreserved autologous bone-marrow-mononuclear-cells (BMMNCs), which contained BMSCs, might maintain bioactivity in vitro, the current study was designed to investigate the effect of cryopreserved BMMNCs before steroid administration for enhancement of bone repair in an established SAON rabbit model. METHODS: 1) In vitro study: 4 weeks before SAON induction in rabbits (n=10), the bone marrow was harvested from iliac crests to isolate BMMNCs (fresh BMMNCs) and then cryopreserved for 8 weeks (cryopreserved BMMNCs). Their activity and osteogenic differentiation were evaluated. Two weeks after SAON induction, the BMMNCs were isolated for the same evaluation. 2) In vivo study: the cryopreserved BMMNCs were implanted into bone tunnel during the core decompression in femora (n=12) after SAON induction. The tissue regeneration was evaluated by micro-CT and histology at 12 weeks after operation. RESULTS: 1) In vitro study: there was no significant difference in activity and osteogenic differentiation between fresh BMMNCs and cryopreserved BMMNCs, but they decreased significantly after SAON induction. 2) In vivo study: bone mineral density (BMD), volume (BV/TV) of bone and the volume and diameter of neovascularization within bone tunnel in BMMNCs treatment group were significantly higher than those in the control group at week 12 after operation. CONCLUSION: The cryopreserved BMMNCs maintained their bioactivities and promoted the bone regeneration and neovascularization within bone tunnel after core decompression in SAON. © 2011 American College of Rheumatology.

SNX-2112, an Hsp90 Inhibitor, Induces Apoptosis and Autophagy Via Degradation of Hsp90 Client Proteins in Human Melanoma A-375 Cells

Cancer Letters. Dec, 2011  |  Pubmed ID: 22182451

SNX-2112 is an Hsp90 inhibitor which is currently undergoing multiple phase 1 clinical trials; however, its mechanism of action needs to be further elaborated. Here we investigated the effects of SNX-2112 in A-375 cells. SNX-2112 induced the degradation of multiple Hsp90 client proteins, activated both the mitochondrial-mediated and death receptor-mediated apoptotic pathways, downregulated Bcl-2 and Bcl-xL, upregulated Bid, cleaved caspase-9, caspase-7, caspase-3 and PARP, and activated caspase-8. The general caspase inhibitor, z-VAD-fmk, did not completely abolish SNX-2112-induced cell death. SNX-2112 induced autophagy in a time- and dose-dependent manner via Akt/mTOR/p70S6K inhibition. SNX-2112 induces significant apoptosis and autophagy in human melanoma A-375 cells, and may be an effective targeted therapy agent.

Antioxidant Phenolic Glycosides from the Bark of Populus Ussuriensis Kom

Natural Product Research. Sep, 2011  |  Pubmed ID: 19662569

Study on the EtOAc soluble fraction from the bark of Populus ussuriensis Kom. resulted in the isolation of three phenolic glycosides, including populoside (1), 7-O-ρ-coumaroylsalirepin (2) and 7-O-caffeoylsalirepin (3), among which 3 is a new compound. The structures of these compounds were elucidated on the basis of spectroscopic evidence. Phenolic glycosides 1-3 exhibited excellent antioxidant activity, evaluated in the ABTS⁺ radical scavenging assay.

Arenobufagin, a Bufadienolide Compound from Toad Venom, Inhibits VEGF-mediated Angiogenesis Through Suppression of VEGFR-2 Signaling Pathway

Biochemical Pharmacology. Jan, 2012  |  Pubmed ID: 22305746

Angiogenesis is crucial for carcinogenesis and other angiogenic processes. Arenobufagin, one of the major components of toad venom, is a traditional Chinese medicine used for cancer therapy. It inhibits cell growth in several cancer cell lines. However, little is known about arenobufagin's anti-angiogenic activity. In this study, we showed that arenobufagin inhibited vascular endothelial growth factor (VEGF)-induced viability, migration, invasion and tube formation in human umbilical vein endothelial cells (HUVECs) in vitro. Arenobufagin also suppressed sprouting formation from VEGF-treated aortic rings in an ex vivo model. Furthermore, we found that arenobufagin blocked angiogenesis in a matrigel plugs assay. Computer simulations suggested that arenobufagin interacted with the ATP-binding sites of VEGFR-2 by docking. In addition, arenobufagin inhibited VEGF-induced VEGFR-2 auto-phosphorylation and suppressed the activity of VEGFR-2-mediated signaling cascades. Taken together, our findings demonstrate that arenobufagin is a specific inhibitor of VEGF-mediated angiogenesis.

Phase Retrieval in Protein Crystallography

Acta Crystallographica. Section A, Foundations of Crystallography. Mar, 2012  |  Pubmed ID: 22338660

Solution of the phase problem is central to crystallographic structure determination. An oversampling method is proposed, based on the hybrid input-output algorithm (HIO) [Fienup (1982). Appl. Opt. 21, 2758-2769], to retrieve the phases of reflections in crystallography. This method can extend low-resolution structures to higher resolution for structure determination of proteins without additional sample preparation. The method requires an envelope of the protein which divides a unit cell into the density region where the proteins are located and the non-density region occupied by solvents. After a few hundred to a few thousand iterations, the correct phases and density maps are recovered. The method has been used successfully in several cases to retrieve the phases from the experimental X-ray diffraction data and the envelopes of proteins constructed from structure files downloaded from the Protein Data Bank. It is hoped that this method will greatly facilitate the ab initio structure determination of proteins.

Fluorinert, an Oxygen Carrier, Improves Cell Culture Performance in Deep Square 96-well Plates by Facilitating Oxygen Transfer

Biotechnology Progress. Jan, 2012  |  Pubmed ID: 21954223

In bioprocess development, the 96-well plate format has been widely used for high-throughput screening of production cell line or culture conditions. However, suspension cell cultures in conventional 96-well plates often fail to reach high cell density under normal agitation presumably due to constraints in oxygen transfer. Although more vigorous agitation can improve gas transfer in 96-well plate format, it often requires specialized instruments. In this report, we employed Fluorinert, a biologically inert perfluorocarbon, to improve oxygen transfer in 96-well plate and to enable the growth of a Chinese Hamster Ovary cell line expressing a recombinant monoclonal antibody. When different amounts of Fluorinert were added to the cell culture medium, a dose-dependent improvement in cell growth was observed in both conventional and deep square 96-well plates. When sufficient Fluorinert was present in the culture, the cell growth rate, the peak cell density, and recombinant protein production levels achieved in deep square 96-wells were comparable to cultures in ventilated shake flasks. Although Fluorinert is known to dissolve gases such as oxygen and CO(2) , it does not dissolve nor extract medium components, such as glucose, lactate, or amino acids. We conclude that mixing Fluorinert with culture media is a suitable model for miniaturization of cell line development and process optimization. Proper cell growth and cellular productivity can be obtained with a standard shaker without the need for any additional aeration or vigorous agitation. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012.