AIM:To develope a reversed-phase high performance liquid chromatographic method for determing chlorogenic acid and caffeic acid in Fanhuncao Granule(Senecio cannabifolius Less). METHODS: The operation was carried out on Eclipse XDB-C_(18) column with the mobile phase consisting of a mixture of acetonitrile-0.4% phosphoric acid(12(∶)88,V/V).The flow rate of 1.0 mL/min and UV detection wavelength at 327 nm were set to determine the contents of chlorogenic acid and caffieic acid. RESULTS: There was good linear relationship between the concentrations and the peak-areas of chlorogenic acid and caffeic acid.The two kinds of standard solutions were both stable in 16 h(RSD=1.55% for chlorogenic acid,1.11% for caffeic acid) The average recovery was 100.1% and 99.8% for chlorogenic acid and caffeic acid,repectively. CONCLUSION: The method is simple,sensitive,rapid and accurate,and can be used for the quality control of Fanhuncao Granule.

Objective To explore the 5,10-methylenetetrahydrofolate reductase( MTHFR) and 5-methyltet-rahydrofolate-homocysteine methyltransferase reductase( MTRR) gene polymorphisms among the Han women in Laiwu City.Methods A total of 559 Han women were recruited.And their oral epithelial cells were collected to extract genome DNA in order to detect gene polymorphisms of MTHFR and MTRR using fluorescence quantitative PCR.Then the results were compared with those in other cities in China.Results The frequency of MTHFR 677CC,677CT and 677TT of Han women in Laiwu city was 14.3%,46.7%and 38.1%,respectively.The frequency of MTHFR 677TT among Laiwu women was significantly different to those of Zhenjiang, Wuhan, Kunming, Deyang, Huizhou, Qionghai (P<0.05).The frequency of AA,AC,CC gene type on MTHFR A1298C was 78.2%,19.7% and 2.1%,respec-tively, the frequency was significantly different to those of Zhenjiang, Wuhan, Kunming, Deyang, Huizhou, Qionghai (P<0.05).The frequency of AA,AG,GG gene type on MTRR A66G was 53.3%,38.8%and 7.9%,respectively. The frequency of MTRR 66GG was significantly different to that of Qionghai(P<0.05).Conclusion The MTHFR, MTRR polymorphism distribution of Han women in Laiwu City has the characteristic of region specificity,respectively.

Objective To explore the genotype distribution of methylenetetrahydrofolate reductase ( MTH-FR)C677T,A1298C and methionine synthase reductase (MTRR) A66G involved in the folic acid biosynthetic path-way among Chinese Han gestational age women in Sanhe City .Methods 601 samples were recruited from Sanhe re-gion,genomic DNA was obtained from the oral mucosa cells .The detections of MTHFR and MTRR gene polymor-phisms conducted with Taqman -MGB technology .The distribution of gene polymorphisms of this study was analyzed and compared with partial regions of China ,which were reported.Results The frequency of MTHFR 677TT among Sanhe women(37.40%) was significantly different to Yanbian (28.30%),Zhenjiang(21.84%),Songzi(15.40%), Deyang(13.80%),Huizhou(10.90%),Qionghai(6.14%),Zibo(43.6%) (χ2 =12.60,87.44,151.95,233.02, 61.87,446.90,7.27,all P<0.05).The frequency of MTHFR 1298CC(2.30%) was significantly different to Zibo (1.44%),Zhenjiang(3.50%),Songzi(2.60%),Deyang(6.26%),Huizhou(7.20%),Qionghai(7.13%) (χ2 =5.84,6.49,14.32,32.54,22.94,53.12,all P<0.05).The frequency of MTRR 66GG (7.50%) was significantly different to Qionghai (9.25%),Songzi(6.40%)(χ2 =16.34,4.10,all P<0.05).Conclusion The MTHFR, MTRR polymorphism distribution of Han women in Sanhe City is region specific ,respective .

AIM: To investigate the antioxidative and antinitrosative effects of copperleaf(Acalypha australis L) on TNBS-induced ulcerative colitis rats. METHODS: Using biochemical method,the activity of antioxidative enzyme SOD,GSH-Px and MDA were determined in normal,model and three rat's groups with low,moderate and high copperleaf decoction respectively.At the same time,the production of NO and the activity of iNOS were also measured.The expression of iNOS mRNA was also determined in normal,model and moderate-copperleaf decoction rats groups through RT-PCR method. RESULTS: The index SOD and GSH-Px increased and MDA reduced significantly in high-and moderate-copperleaf decoction groups compared with the model group.The production of NO the activity of iNOS reduced significantly in high-and moderate-copperleaf decoction groups compared with the model group.RT-PCR results demonstrated that the expression of iNOS mRNA was significantly inhibited in TNBS-induced ulcerative colitis rats after being treated with moderate-dosage copperleaf. CONCLUSION: Acalypha australis L.has antioxidative and antinitrosative effects which is probably one of the mechanism of copperleaf for treating UC.

Objective:To prepare injectable interferon ?-2b(IFN-?-2b) loaded microsphere and develop a long-acting dosage form.Methods: IFN-?-2b loaded microspheres were prepared with poly(lactic-co-glycolic acid)(PLGA) as carrier material by double emulsion(w/o/w) method and solid in oil in oil(s/o/o) method separately.Physical and chemical characteristics of microspheres(mean diameter,morphology and drug entrapment efficiency) were evaluated;the in vitro release behavior and influencing factors of the microspheres were determined by micro-BCA(bicinchoninic acid) method;and IFN-?-2b stability during encapsulation and in vitro release was evaluated by sodium dodecyl sulfate polyacrylamide gel electropheresis.Results: The 2 types of microspheres produced had good shape and dispersive quality and a drug entrapment efficiency of more than 80%.IFN-?-2b bulk ultrafitration can significantly influence the mean diameter and in vitro release behavior of microspheres prepared by w/o/w method.The accumulated release(within 1 month) of the microspheres prepared by both methods was significantly improved when using PLGA with lower inherent viscosity.SDS-PAGE test showed aggregation of IFN-?-2b with s/o/o method,while there was no difference between the electrophoretic behavior of bulk IFN-?-2b and IFN-?-2b in microspheres prepared by w/o/w method.Conclusion: IFN-?-2b can be encapsulated into injectable microspheres to yield a one-month continuous release by both w/o/w method and s/o/o method.

Objective To discuss the correlation between MTHFR gene polymorphism and unexplained recurrent spontaneous abortion.Methods A case control study was used in this study,140 patients with unexplained recurrent spontaneous abortion(UR-SA) (abortion group)and 143 cases of normal women(control group)were recruited.Genomic DNA was obtained and extracted from the oral mucosa cells.Fluorescence quantitative PCR was used to examine the MTHFR gene polymorphisms,and Taqman-MGB technology was conducted to analysis the relationship between single nucleotide polymorphism and disease.Results There was statistically significant difference in the frequencies of C677T genotype and alleles between the two groups(P＜0.05).However,no significant difference in the frequencies of A1298C genotype and alleles between the two groups(P＞0.05).Conclusion MTHFR gene C677T polymorphism might be one of the genetic risk factors of URSA.

Objectives To investigate the relationship of methylenetetrahydrofolate reductase (MTHFR) and methioninesynthase reductase (MTRR) with unexplained recurrent spontaneous abortion (URSA). Methods Case control study was used to select 244 patients with URSA (miscarriage group) and 116 normal women (control group) who were admitted to Tianjin Medical University General Hospital and Tianjin Women’s and Children’s Health Center from January 2013 to March 2015. The oral mucosal epithelial cells were extracted using fluorescence quantitative PCR to detect MTHFR gene C677T, A1298C and MTRR gene loci of A66G single nucleotide polymorphisms (SNP). The relationship between folate metabolism related gene polymorphisms of MTHFR and MTRR and URSA was analysed. Results The frequency of C677T genotype MTHFR was significantly higher in URSA group than that in the control group, and the frequency of CT genotype was significantly lower than that of the control group (P<0.05). There was no significant difference in the frequencies of A1298C MTRR and A66G MTHFR between the two groups. The activity of MTHFR, red cell folate and plasma folate levels were significantly lower in URSA group than those of control group. Homocysteine levels were significantly higher in URSA group than those of control group (P<0.05). There were no significant differences in serum folic acid, red cell folate, homocysteine cysteine levels between patients <35 years old and ≥ 35 years old in URSA group. Conclusion C677TMTHFR gene polymorphism is associated with unexplained recurrent spontaneous abortion.

Objective To investigate the distribution characteristics and risk factors of intracranial atherosclerotic stenosis ischemic stroke. Methods We retrospectively collected 342 consecutive patients with first-ever ischemic stroke. Clinical data was collected including demographics, the presence of risk factors,MRI with MRA and other routine admis?sion laboratory tests. Results Intracranial atherosclerotic stenosis (ICAS) was located most frequently in MCA (47.0%), Extracranial internal carotid artery was the most common affected artery (65.0%) among extracranial atherosclerotic steno? sis (ECAS). MetS (OR=1.586,95%CI:1.232~2.268), ApoB/ApoA1 ratio (OR=1.926,95%CI:1.051~4.288), were as?sociated with ICAS (vs ECAS), whereas hypertension (OR=3.603,95%CI:1.675~12.485), MetS (OR=2.268,95%CI:1.274~6.103), HbA1c (OR=2.015,95%CI:1.182~5.613) and ApoB/ ApoA I ratio (OR=1.948,95%CI:1.157~4.285) were related to ICAS (vs NCAS). Hypertension (OR=2.437,95%CI:1.492~3.505,P=0.005), Hcy (OR=2.437,95%CI:1.492~3.505,P=0.005) and HbA1c (OR=1.769,95%CI:1.034~3.121, P=0.005) were the independent risk factors re?lated to posterior circulation strokes (vs anterior circulation strokes ) in ICAS strokes. Conclusions The occurrence of ICAS may be more frequent than that of ECAS in ischemic stroke. Posterior circulation ICAS strokes seems to be close?ly associated with metabolic derangement.

Objective To investigate the influence of the genotype distribution of methylenetetrahydrofolate reductase (MTHFR) C677T,A1298C and methionine synthase reductase (MTRR) A66G in threatened abortion of Chinese Han gestationalage women in Sanya city,which involved in the folic acid biosynthetic pathway among.Methods One hundred and thirty-nine samples of case group and the same number of control group were recruited from Sanya region in Hainan Province.Genomic DNA was extracted from the mucosal epithelium of the subjects.The gene polyrnorphisms of MTHFR and MTRR were detected by Fluorescence quantitative PCR technology.The distribution frequencies of both case group and control group.were analyzed and compared,to investigate the effect of the gene polymorphisms on threatened abortion.Results Both the case group and the control group complied with Hardy-Weinberg law.The genotype frequency of MTHFR C677T,MTHFR A1298C and MTRR A66G were not significantly different.Conclusion This study suggests that the gene polymorphism which involved in folic metabolism was not significantly different from the group of threatened abortion and the control group,and whether the metabolism related genes are the risk factors of threatened abortion need to be further discussed.

Objective To evaluate the effects of folic acid supplement on subjects with different 5, 10-methylenetet-rahydrofolate reductase (MTHFR) genotypes. Methods One hundred and eleven healthy women were divided into CC, CT and TT groups according to their MTHFR C677T genotypes. In each group subjects were randomly sub-divided into interven-tion (400 μg/d folic acid supplement) and control (usual diet) groups. The plasma folate, red blood cell (RBC) folate and plasma homocysteine (Hcy) concentration were measured at baseline and two months after intervention. Results The plasma folate was lower and the plasma Hcy was higher in the TT genotype than those in CC or CT genotypes (P＜0.05 or P＜0.01). After two months of intervention, the levels of plasma folate, RBC folate concentration increased while the plasma Hcy concen-tration decreased in all three intervention groups. Although the plasma folate concentration increased the most obvious in TT genotype than that of CC and CT genotypes, P＜0.05), the plasma Hcy concentration decreased the most obvious in TT geno-type than that of CT genotype, P＜0.05). Logistic regression analysis showed that the MTHFR TT genotype was a risk factor of high Hcy concentration, which was 8.078 times compared with that of CC genotype (P＜0.05). Conclusion Folic acid sup-plement can significantly increase plasma folate and red cell folate concentration, and reduce plasma Hcy concentration in all MTHFR genotypes. TT genotype was the most dangerous in disorder of folic metabolic and high Hcy concentration. However, low-dose folic acid supplement cannot reduce the risk of high Hcy concentration.