Objective To study the effect of thrombin on proliferation and invasion of esophageal cancer cell line Eca109, and to explore its possible mechanism. Methods The proliferation and invasion of Eca 109 cells treated with thrombin were detected by MTT and Transwell assay, respectively. The activity of matrix metalloproteinase 2 (MMP-2) and MMP-9 in the supernatant of Eca109 cells was detected by gelatin zymography. Reverse transcription polymerase chain reaction (PCR) and immunocytochemistry were used to study the mRNA expression of protease-activated receptor 1 (PAR-1), the important receptor of thrombin, and subcellular localization of PAR-1 protein in Eca109 cells, respectively. Results Thrombin could promote Eca109 cells proliferation in a dose-dependent manner. Cell proliferative rates of 0.5 U/ml and 1.0 U/ml thrombin were 34.38 % and 57.19 %, respectively (P< 0.05). Compared to that of control group, the number of Eca109 cells incubated with 1.0 U/ml thrombin invading through the basement membrane of Transwell was increased (303.33 ±6.66 vs. 116.33 ±11.51, P< 0.05). When treated with various concentrations of thrombin for 24 h, the activities of MMP-2 and MMP-9, especially MMP-9, in the supernatant of Eca109 cells were increased in a dose-dependent manner. Eca109 cells expressed PAR-1 mRNA, and PAR-1 protein was mainly located on the cellular membrane. Conclusion Thrombin increases proliferation and invasion of esophageal cancer Eca109 cells and enhances the activities of MMP-2 and MMP-9 in cells supernatant, which might be induced by activation of PAR-1 located on cellular membrane.

AIM: To explore the optimal separation of the total flavonoid from Radix Puerariae by selecting appropriate macroporous resins and to study systematically the factors which affect separation. METHODS: Static and dynamic adsoption desorption methods were adopted, and evaluated for separating efficiency by measuring the concentration of total flavonoid in Radix Pueraria with UV spectrophotometer. RESULTS: The SP70 macroporous resin had the best separating efficiency when the flavonoid content in the liquid was 0.5g per 1mL equivalemt to raw material. pH5～6. the volume of drug 60BV (resin bed volume) with the adsorption power 2BV/h. and the volume of 70%(mL?mL -1 ) ethanol as eluant 4BV with desorption power 2BV/h. In result. total flavonoid's purity could reach 80%. CONCLUSION: This method is simple and feasible with good effect of separation, which can meet industrial requirements.

AIM: To determine coumarin and flavonoid in Compound Buguzhi Sustained-Release Tablet(Fructus Psoraleae,etc). METHODS: HPLC method was developed.The separation was performed on an ODS column with the mobile phase of CH_3OH-H_2O.The flow rate was 1 mL/min and the detection wavelength was 320 nm. RESULTS: The result showed that there was good linear relation between the area and the concentration of psoralen、isopsoralen、isobavachin、bavachin、corylin、corylifolinin、bvachinin、psoraidin and bavachalcone within 0.4-152.0,26.4-132.0,13.6-66.0,27.2-136.0,31.2-156.0,48.8-244.0,50.4-252.0,36.0-180.0,(8.8)-44.0 ?g/mL.The average recovery of psoralen and bavachin is 98.7% and 97.6%. CONCLUSION: This method is convenient,reliable,and can be used to control the quality of Compound Buguzhi Sustained-Release Tablet.

Objective To investigate the effect of Fructus Psoraleae on the proliferation and differentiation of cultured osteoblast isolated from neonatal rat's calvarium.Methods Calvarium osteoblasts of new born SD rat were cultured and exposed to different concentrations of Fructus Psoraleae extract,then the content of alkaline phosphatase(ALP)in the cell was measured and the cell proliferation was detected by tetrazoline colorimetry(MTT).Results Ethyl acetate extract and alcohol extract from Fructus Psoraleae can improve the activity of ALP and the proliferation of rat's cultured osteoblasts,the optimum concentration was 2.0?10-3 mg/mL.Conclusion Ethyl acetate extract and alcohol extract from Fructus Psoraleae can stimulate the differentiation and proliferation of osteoblasts in vitro.

Objective To screen out the effective constituents with estrogenic action in Fructus psoraleae. Methods Fructus psoraleae was separated by using systemic solvent distillation method, and the effective fraction was screened out by the experiment of increasing the uterine weight in mice. Then the petroleum fraction was separated by sil; ca gel chromatrography and the effective constituent was screened by the experiment of increasing the uterine weight in mice. Results High-dose(5g/kg) of the petroleum fraction showed obvious estrogenic action (P

Objective To explore the association between SLC2A9,SLC17A3,ABCG2 single nucleotide polymorphisms and gout susceptibility in Quanzhou.Methods One hundred and fifty-four cases of gout patients and 160 healthy controls were selected,single nucleotide polymorphisms (SNP) of SLC2A9 SLC17A3,ABCG2 with tri-primer polymerase chain reaction (PCR) were tested and the relation between different genotypes and primary gout prevalence were analyzed.Results High risk genotype frequency of rs16890979 was 93.5％ and 70.0％ in patients and healthy people,respectively (the difference of genotype frequency between the two groups was statistically significant (x2=55.377,P＜0.01).High risk allele frequency was 79.9％ and 48.4％ in patients and healthy people,respectively (allele frequency in different population was statistically significant,x2=67.128,P＜0.01).High risk genotype frequency of rs2231142 was 68.8％ and 38.7％ in patients and healthy people,respectively (the difference of the genotype frequency was statistically significant,x2=29.129,P＜0.01);High risk allele frequency was 43.5％ and 23.4％ in patients and healthy people,respectively (the difference of allele frequency was statistically significant,x2=28.468,P＜0.01) ; rs1165205was a protective SNP,low risk genotype frequency was 42.2％ and 45.6％ in patients and healthy people,respectively (the difference of genotype frequency was statistically significant,x2=0.373,P=0.571); High risk allele frequency was 26.0％ and 28.1％ in patients and healthy people,respectively (the difference of allele frequency was not statistically significant,x2=0.270,P=0.364).Conclusion SNP loci rs16890979 of SLC2A9 gene and rs2231142 of ABCG2 gene can be used as genetic markers for primary gout susceptibility in the Quanzhou area,but SNP loci rs1165205 of SLC17A3 gene has little correlation with the prevalence of primary gout in Quanzhou residents.

Objective Hirsutella sinensis (HS) is the anmorph of Ophiocordyceps sinensis (Cordyceps sinensis). O.sinensis and Panax notoginseng are two popular Chinese herbs, commonly used in traditional Chinese prescriptions for the treatment of various diseases. A combination of HS extract with P. notoginseng saponin (PNS) extract demonstrated more prominent lung-protective activity than the two herbs individually used in our preliminary studies. This study further investigated the action of their combination (HSPNS) on anti pulmonary fibrosis using a Bleomycin (BLM)induced mouse model. Methods BLM-treated Kunming mice was given HSPNS daily for 7, 14 or 28 d via ig administration. After treatment, following parameters were monitored using proper methods, respectively. Lung index, serum and lung malondialdehyde (MDA) and hydroxyproline (HYP) contents, lung superoxide dismutase (SOD) activity, transforming growth factor β1 (TGF-β1), and expression levels of collagen Ⅰ (Col- Ⅰ) and collagen Ⅲ (Col-Ⅲ). The lung biopsies were also dissected for semiquantitative histological analysis. Results The results indicated that HSPNS significantly reduced lung index, MDA and HYP contents, and expression levels of TGF-β1,Col- Ⅰ, and Col-Ⅲ. The combination also remarkably enhanced SOD activity compared with BLM-induced group.Moreover, the severe pulmonary fibrosis histopathological changes induced by BLM could be attenuated by HSPNS treatment. Conclusion These results suggest that HSPNS could significantly inhibit the progression of pulmonary fibrosis induced by BLM and its inhibitory effect might associate with its ability to scavenge free radicals, decrease TGF-β1 level, and inhibit collagen synthesis.

Objective To improve the tri-primer allele gene amplification for realizing the single nucleotide polymorphisms (SNP) genotyping of the peripheral blood sample .Methods Aiming at the peripheral blood samples with the clinical usual antico-agulation processing by EDTA ,heparin and citrate ,with the locus rs1165205 as the target site ,the buffer solution(YW) suitable for whole blood was prepared and the PCR amplification system and the amplification condition were optimized for realizing the detec-tion of SNP genotyping .Results The genotyping results of locus rs1165205 by improved tri-primer allele gene amplification method were consistent with the results of the Sanger sequencing method ,and the peripheral blood samples treated by different anticoagu-lant were genotyped by the improved tri-primer ASA .Among 80 samples ,various genotypes of locus rs1165205 had no statistical differences in the distribution between the gout population and non-gout population(P= 0 .335) .Conclusion The improved tri-primer allele gene amplification method can be adopted to conduct the rapid genotyping research on gout SNP locus of the peripheral blood samples with the clinical usual anticoagulation processing .

Objective A very high prevalence of rheumatoid arthritis (RA) is observed in Minnan population in China.We aimed to explore the genetic characteristics of RA in Minnan population and genetic mechanisms of RA by studying the associations of three single nucleotide polymorphisms (SNPs) of signal transducer and activator of transcription 4 (STAT-4) (rs7574865),the cytotoxic T-lymphocyte antigen-4 (CTLA)-4 (rs3087243) and chromosome 9p21.3(rs1333049) with RA in Minnan population.Methods A case-control study of 119 RA patients and 125 normal controls from Quanzhou were enrolled.SNPs (rs7574865,rs3087243,rs1333049) were genotyped by allele-specific polymerase chain reaction (PCR) and analyzed by SPSS 18.0.x2-test was applied to compare allele and genotype frequeneies betweeen cases and controlsLogistic regression models were used to analyze the SNPs.Results The results showed the genotype distributions of STAT4 genes were significantly different between case and control groups (P＜0.01).Compared with the GT heterozygous genotype,TT and GG homozygosity carriers had a lower risk (OR=0498 and 0.300,P=0.018 and P=0.002 respectively).There was not statistical difference in genotypes and allele in CTLA-4 (rs3087243) between RA patients and healthy controls (x2=4.083,P=0.130),but compared with the AG genotyoe,GG homozygosity carriers had a lower risk on basis of statistics (OR=0.580,P=0.04).There was not statistical difference in genotypes and allele in the chromosome 9p21.3 (rs1333049) (P＞0.05),but compared with the GG genotype carriers,CC and GC genotypes carriers had a lower risk on basis of statistics (OR=0.565,P=0.0495).Conclusion Chromosome 9p21.3 (rs1333049) and CTLA-4 rs3087243 G/A may not be associated with susceptibility to RA in Minnan popula-tions.This replication study confirmes that STAT4 rs7574865 G/T polymorphism is associated with susceptibility to RA in Minnan population.

Objective: To analyze the clinical characteristics of hypophosphatemic osteomalacia induced by adefovir dipivoxil (ADV) in order to improve the understanding of the disease.Methods: A retrospective analysis was performed according to the medical records of 11 cases of ADV-induced hypophosphatemic osteomalacia.The medical history, laboratory indicators (ALT, AST, ALB, SCr, UA, blood glucose, blood pH, BE), bone metabolic markers (25OHD3, PTH, tP1NP, β-CTX, OC), urine indicators (urine pH, 24h urine Ca, 24h urine P, 24h urine Pro, urine Scr), DXA and skeleton ECT signs of the patients with hypophosphatemic osteomalacia induced by ADV were analyzed, and the symptoms, blood P, AKP level and urine routines were followed up after 1-month withdrawal and in July, 2016, respectively.Results: The mean ADV administration time of the 11 patients was (5.7±1.2) years, and the bone pain time was (2.2±0.6) years.The serum P was (0.45±0.99)mmol·L-1, 24h urine P was (17.9±4.8)mmol, AKP was (248±107)IU·L-1,the concentration threshold of renal phosphate was(0.31±0.10)mmol·L-1.After the one-month withdrawal of ADV, the bone pain in the patients were all relieved, and with the phosphorus supplement, the level of serum phosphorus was increased.In July of 2016, the average withdrawal time of ADV was (18.3±10.7) months, the serum phosphorus significantly increased and AKP significantly decreased when compared with that on the admission and 1 month after the ADV withdrawal (P<0.05), and the serum phosphorus of 2 patients returned to normal with the recovery rate of 20% (2/10).The regression analysis showed that the influencing factors on serum phosphorus on the admission were renal concentration threshold of phosphate and tP1NP (P<0.05);the influencing factor on serum phosphorus on the last follow-up was bone mineral density at the admission (P<0.05).Conclusion: Hypophosphatemic osteomalacia is a potential side effect of ADV, and ADV-induced renal injury is not completely reversible, which should be paid more attention in clinical work.