A in vitro platelet aggregation bioassay was developed for the quality control of XST capsules. The in vitro anti-platelet aggregation effect in rats was observed to detect the bioactivity of XST capsules. Panax notoginseng saponins and Xuesaitong lyophilizedpowder for injection were taken as standard control substances to determine the potency. According to the results, XST capsules showeda significant inhibitory effect on thrombin-induced platelet aggregation in a dose-dependent manner. The in vitro anti-platelet activity oflyophilized powder for injection was stabler than that of Panax notoginseng saponins, and so suitable to serve as a standard control substance. The biological potency of XST capsules compared with standard control substance was detected by using parallel line assay. According to the results, the established bioassay method had a good repeatability (RSD 2.92%). The sample test results could pass thereliability test(linear deviation P > 0.05, parallel deviation P > 0.05). This bioassay method could be used as one of the complementary quality control methods for XST capsules.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Panax notoginseng ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

The efficient and safe delivery of drugs to the therapeutic site through the biofilm has traditionally been a difficult and hot topic in the field of drug delivery. In recent years, alkyl polyglycoside (APG) have become ideal penetration enhancers for drug delivery systems because of their high permeability, good safety and biodegradability, which has attracted wide attention of domestic and foreign researchers. In this paper, the physical and chemical properties, characteristics, action mechanism and application of APG in drug delivery system are reviewed, and its application prospect in drug delivery system is prospected.

OBJECTIVEThe B7-H1/PD-1 co-signaling pathway has recently been found to play a pivotal role in the immune evasion of tumor cells from host immune system. The aim of this study was to examine the B7-H1 and PD-1 expression and TILs status in gastric cancer and to elucidate the clinical relevance of B7-H1 and PD-1 to the pathogenesis of gastric carcinoma.

METHODSImmunohistochemistry and ANAE histochemical staining were used to investigate the in situ expression of B7-H1 and PD-1 and TILs status in the gastric tissues. RT-PCR was used to explore B7-H1 and PD-1 expression at the transcriptional level. The B7-H1 expression at protein level was detected by Western blot.

RESULTSExpression of B7-H1 and PD-1 was found to be increased in gastric carcinoma, but absent in normal gastric tissue. B7-H1 expression in gastric carcinoma was inversely correlated with TILs infiltration. B7-H1 but not PD-1 expression in tumor tissue was significantly correlated with some clinicopathhological variables including depth of invasion, lymph node metastasis and distant metastasis.

CONCLUSIONB7-H1 and PD-1 expressions are increased in gastric carcinoma. This signaling pathway may inhibit antitumor immune responses in gastric carcinoma. B7-H1 expression plays a critical role in the pathogenesis of human gastric carcinoma,and might be a promising prognostic marker and therapeutic target in the treatment of this disease.

Adult ; Aged ; Antigens, CD ; genetics ; metabolism ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; B7-H1 Antigen ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Lymphatic Metastasis ; Lymphocyte Subsets ; immunology ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Programmed Cell Death 1 Receptor ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; genetics ; immunology ; pathology

OBJECTIVETo investigate the prevalence and subtypes of microdeletions in azoospermia factor (AZF) region in infertile men from Sichuan in order to correlate genotypes with phenotypes.

METHODSMultiplex-PCR was used to detect sequence tagged sites (STS) of AZF microdeletions in 1011 infertile men including 713 cases of non-obstructive azoospermia and 298 cases of severe oligospermia.

RESULTSThe overall prevalence of microdeletions was 10.48% (106/1011), and the deletion rates were 11.08% (79/713) in non-obstructive azoospermia and 9.06% (27/298) in severe oligospermia. Complete AZFa or AZFb deletions were associated with azoospermia, whereas AZFc deletion (60.38%) was the most frequent deletion. The deletions were associated with variable spermatogenic phenotypes, and 37.50% of the patients with a deletion had sperms in the ejaculate. A mild decline in sperm concentration was found in two cases with partial AZFb deletion and one case with partial AZFb-c deletion.

CONCLUSIONDeletions of the AZFc region were most commonly found in our patients. All cases with complete AZFa or AZFb deletions and a proportion of cases with AZFc deletion were associated with azoospermia. Our study has provided more insight into the genotype-phenotype correlation, and confirmed that Yq microdeletion screening has a significant value for the diagnosis for male infertility.

Adult ; Azoospermia ; genetics ; Chromosome Deletion ; Chromosomes, Human, Y ; Genetic Association Studies ; methods ; Humans ; Infertility, Male ; genetics ; Male ; Phenotype ; Young Adult

OBJECTIVETo evaluate whether or not administration of folic acid and resveratrol have preventive effects on cleft palate formation as well as the comparison of the two drugs' s effects.

METHODSPregnant mice were randomly divided into 9 groups, with 8 mice in each group. The TCDD group mice were dosed with TCDD 28 microg/kg body weight on gestation day 10 (GD 10) animals in folic acid group were respectively dosed with folic acid 15, 10, 5 mg/kg and TCDD 28 microg/kg; resveratrol treated mice were divided into 3 groups: resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13 in resveratrol (GD8-13 ) group; resveratrol 50 mg/kg were orally administered for 6 consecutive days, from gestational day GD 8 to GD13, followed hy an oral administered with TCDD on GD10 in resveratrol (GD8-13) + TCDD group; resveratrol 50mg/kg and TCDD 28 microg/kg were used by gavage administration at GD10 in resveratrol (GD10) + TCDD group. Control mice were treated with the same volume of water for 6 consecutive days from GD8 to GD13 and were given a single dose of corn oil on GD10. The pregnant mice weight and embryos, the number of live, cleft palate, dead and resorption fetal mice were recorded on GD 17.5. The coronal sections of the fetal mice heads were prepared at GD 17.5 and observed by microscopy.

RESULTSTotal frequency of clefts was 92.86% in TCDD group, 84.00% (15 mg), 73.08% (10 mg), 84.00% (5 mg) in folic acid + TCDD groups, 0% in resveratrol (GD10) group, 74.51% (GD10), 57.78% (GD8-13) in resveratrol + TCDD groups. The frequency of cleft was 0% in the control group. Compared with the control and the TCDD groups, there were significant differences in the number of live, dead and resorption fetal mice in TCCD + resveratrol (GD8-13) group (P < 0.05). No significant differences in embryonic weight, live fetuses weight, the number of live, dead and resorption fetal mice were found in the other groups (P > 0.05).

CONCLUSIONTest dose of folic acid and resveratrol both had certain antagonistic effect on cleft palate in mice induced by TCDD, with folic acid 10 mg/kg, resveratrol 50 mg/kg GD8-13 doses having stronger antagonistic action. Effects of both the two drugs have no significant difference, but resveratrol (50 mg/kg, GD8-13) significantly affects the fetal mice's growth and development under TCDD exposure in utero.

Abnormalities, Drug-Induced ; prevention & control ; Animals ; Cleft Palate ; chemically induced ; prevention & control ; Female ; Fetus ; Folic Acid ; administration & dosage ; pharmacology ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; antagonists & inhibitors ; Pregnancy ; Random Allocation ; Stilbenes ; administration & dosage ; pharmacology ; Teratogens

Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; blood ; diagnosis ; Female ; Gene Amplification ; Humans ; Insulin-Like Growth Factor II ; analysis ; genetics ; Liver Neoplasms ; blood ; diagnosis ; Male ; Prognosis ; RNA, Messenger ; blood

Objective To understand the prevalence of anemia and its influencing factors in infants aged 6 to 24 months in poor areas of Gansu Province, and to provide reference for improving the prevalence of anemia in local children. Methods A multi-stage sampling method was used to conduct a survey on infant and caregivers, in the 12 children nutrition improvement project counties in Gansu Province. T-test and analysis of variance were used to compare the hemoglobin content, the chi-square test was used to compare the prevalence of anemia, and the Logistic regression model was used to analyze the influencing factors. Results Among the 3 188 effective data, the detection rate of anemia was 25.69% (819). The detection rate of anemia among boys and girls was 24.54% and 26.90%, respectively. There was no significant difference in the detection rate of anemia among different sexes ( 2=2.326, P=0.127). The detection rate of anemia between different age groups were statistically significant ( 2=42.339, P<0.001); The results of multivariate analysis showed that children's age, children's ethnic groups, parents' awareness of feeding knowledge, the feeding method of 6 months after birth and the way of taking nutritional packs were associated with anemia (all P<0.05). Conclusions The prevalence of anemia in infants aged 6 to 24 months in poor areas in Gansu Province was relatively high. Strengthening the education of caretakers’ knowledge of guardian feeding and scientific child-rearing, and ensuring the nutrition packages intake can significantly reduce the prevalence of anemia prevalence in poor areas in Gansu Province.

OBJECTIVETo focus on the study of the effect on proliferation and apoptosis of human aortic smooth muscle cells (ASMC) by adeno-associated virus (AAV) vector carrying antisense thrombin receptor (ATR) and p21 double gene co-expression system.

METHODSCultured human AMSC was infected with recombinant AAV containing ATR, p21 single gene and AP double gene respectively. The integration and expression of genes were confirmed by semi-quantitative RT-PCR. The anti-proliferation effect was determined by MTT assay. Cell cycle and apoptotic cell counts were measured through Flow Cytometry. The rate of apoptotic cells was examined with acridine orange/ethidium bromide(AO/EB) stain.

RESULTSRT-PCR indicated that the exogenous genes had been integrated into ASMC. The rates of cell survival were decreased by 16.67%, 21.60%, and 29.4% and the cell counts of G0/G1 phase were (61.8 +/- 2.9)%, (82.5 +/- 4.0)%, (80.4 +/- 6.1)% in ATR, p21 and AP group respectively after rAAV infected 4 days. The level and area of apoptotic peak were greater in AP double gene than ATR and p21 single gene. Cell stain indicated that apoptotic cells were (7.2 +/- 3.3)%, (10.7 +/- 5.6)%, and (18.3 +/- 2.7)% in each transgene group compared with (1.5 +/- 0.8)% in control group.

CONCLUSIONAP double gene co-expression system has powerful effect for inhibiting proliferation and inducing apoptosis ASMC than ATR and p21 single gene and that is a superior way for gene therapy to restenosis.

Adenoviruses, Human ; genetics ; Antisense Elements (Genetics) ; Aorta ; cytology ; Apoptosis ; Cell Division ; Cells, Cultured ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; biosynthesis ; genetics ; Fetus ; Genetic Vectors ; Humans ; Muscle, Smooth, Vascular ; cytology ; Receptors, Thrombin ; biosynthesis ; genetics

OBJECTIVETo explore the mechanism of cleft palate in mice induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD).

METHODSOn gestation day 10 (GD 10), 12 pregnant mice were randomly divided into two groups as the treated group and the control group with 6 mice in each group. The mice in the treated group received intragastric administration with 64 microg TCDD/kg, while the mice in the control group received equivalent corn oil. The embryos were examined under stereomicroscope to detect the incidence of cleft palate on GD 18.5. Another 18 pregnant mice were randomly divided into two groups (treated group and control group) on GD 10 with 9 pregnant mice in each group. Then each group was divided into 3 subgroups: GD 13.5, GD 14.5 and GD 15.5, with 3 pregnant mice in each subgroup. The palatal shelves were dissected from the embryos for RNA and DNA extraction on GD 13.5, GD 14.5 and GD 15.5. At last the expression of Smad 2-4 and Smad 7 mRNA was investigated by RT-PCR, and the TGF-beta3 promoter methylamine levels were investigated by methylation specific PCR (MSP).

RESULTSThe cleft palate mice model was established successfully by exposing pregnant C57BL/6J mice to TCDD. Total frequency of clefts was 100% in TCDD group, and the frequency of clefts was 0 in the control group. The relative expression of Smad 2 mRNA was 0.263 +/- 0.088, 0.296 +/- 0.016 and 0.159 +/- 0.027 in TCDD group, 0.180 +/- 0.042, 0.282 +/- 0.029 and 0.165 +/- 0.018 in control group. The relative expression of Smad 3 mRNA was 0.453 +/- 0.153, 0.551 +/- 0.160 and 0.328 +/- 0.049 in TCDD group, 0.375 +/- 0.126, 0.510 +/- 0.145 and 0.259 +/- 0.035 in control group. The relative expression of Smad 4 mRNA was 0.675 +/- 0.174, 0.577 +/- 0.070 and 0.396 +/- 0.066 in TCDD group, 0.557 +/- 0.138, 0.587 +/- 0.080 and 0.441 +/- 0.054 in control group. The relative expression of Smad 7 mRNA was 0.283 +/- 0.050, 0.320 +/- 0.068 and 0.169 +/- 0.045 in TCDD group, 0.207 +/- 0.043, 0.288 +/- 0.051 and 0.155 +/- 0.040 in control group. There was no significant difference between the TCDD treated mice and the control (P > 0.05). The TGF-beta3 promoters were at the un-methylation state both in the TCDD treated and control group.

CONCLUSIONIt suggests that TCDD could induce a stable formation of cleft palate, but it is not through the TGF-beta/Smad signaling nor through the modification of TGF-beta3 promoter methylation.

Animals ; Cleft Palate ; chemically induced ; DNA Methylation ; Female ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; toxicity ; Pregnancy ; Promoter Regions, Genetic ; Signal Transduction ; Smad Proteins ; metabolism ; Teratogens ; toxicity ; Transforming Growth Factor beta3 ; metabolism