Objective To elucidate the mechanism of angiotensin (Ang) Ⅱ in regulation of renin synthesis and secretion in primary cultured juxtaglomerular granular cells (JGCs). Methods Mice JGCs were isolated and cultured as described before. Real-time PCR was used to demonstrate the expression of angiotensin converting enzyme (CE) and angiotencin receptor (AT1, AT2) in JGCs. Ang Ⅱ was co-cultured with JGCs stimulated by PGE2 and isoproternol or not. Renin activity in supernatant was detected by radioimmunoassay and renin mRNA expression was examined by real-time PCR. Different concentrations of Ang Ⅱ were co-cultured with JGCs with different time (1 h, 4 h and 24 h), cAMP in cell lysate and supernatant was measured by Cayman Cyclic AMP EIA Kit. The cytoplasmic calcium was decreased by BAPTA-AM, and increased by thapsigargin and cyclopiazonic acid, which could be used to observe the cAMP concentration affected by calcium. Adenylyl cyclase type 5, 6 (AC5, AC6) mRNA expression of JGCs co-cultured with Ang Ⅱ was measured by real-time PCR. Results Real-time PCR confirmed the expression of ACE and AT mRNA in JGCs of WT mice. Angiotensin Ⅱ reduced renin secretion in primary cultures of JGCs [(370.6±36.9) vs (299.6±25.7) ng AngI'ml~(-1)·h~(-1), P=0.014]. Angiotensin Ⅱ dose-dependently down-regulated forskolin-stimulated cAMP production in JGCs. Thapsigargin and cyclopiazonic acid decreased cAMP level. BAPTA-AM increased cAMP production obviously [(11.09±0.48) vs (3.55±0.47) nmol/L, P<0.01]. Ang Ⅱ inhibited AC5 mRNA expression by 42.12%, but did not inhibit AC6 mRNA expression. Conclusions Angiotensin Ⅱ can directly inhibit renin synthesis and secrestion maybe through reducing AC-stimulated cAMP levels. Crosstalk between calcium and cAMP system may exist in precise regulation of renin in JGCs.

Objective To evaluate the effectiveness of multiple quantitative fluorescence PCR ( QF-PCR) as a rapid technique for prenatal diagnosis of common chromosome aneuploidies , in order to optimize the prenatal diagnosis and shorten the period of diagnosis.Methods Totally 731 amniotic fluid samples of pregnant subjects ,who were referred to the Women′s Hospital School of Medicine Zhejiang University during August 2013 and September 2015, were analyzed with conventional karyotype and the QF-PCR technique by short tandem repeat(STR) markers to detect chromosomes 13,18,21,X and Y aneuploidies.There were 558 samples detected by single blind method , 173 samples detected by double blind method.Results All of the 731 amniotic fluid samples were tested in this study by QF-PCR and the results were compared to the conventional cytogenetic analysis results of the same sample.Totally 558 samples with single blind method detected 5 trisomy 21, 2 trisomy 18, 1 trisomy 13, 1(45,X), 1(47,XXY), 1(47,XYY), 1(47,XXX) and 1(69,XXX), 173 samples with double blind method detected 1 trisomy 21 and 1 trisomy 18.The rapid QF-PCR assay was successful to detect all aneuploidies involving chromosomes 21, 18, 13, X and Y in prenatal diagnosis , which were verified by chromosome karyotype analysis.The results of QF-PCR method were compared with the results of chromosome karyotype analysis , the positive rate was 15/16, the negative rate was 100%(715/715).Non chimeric chromosome abnormality detection rate was 15/15.Conclusions The multiple QF-PCR was a reliable method of detecting common chromosome aneuploidies for rapid prenatal diagnosis.As an important supplement of karyotype analysis , it was of great significance to optimize and improve the prenatal diagnosis system , and might provide more appropriate diagnostic methods for pregnant women.

Objective To discuss the variation range and the indication of soluble serum Fas (sFas) in aplastic anemia (AA) patients.Methods The level of sFas receptor in 38 AA patients was assayed by ELISA,with 30 cases as controls.Results The level of sFas receptor in AA patients was apparently lower than those of the control group (P

Objective To evaluate the clinical result of system rehabilitation training on joint rehabilitation after operation in cruciate ligament injury of the knee joint patients.Methods All patients were followed up for 6～18 months(average 12.8 months).The clinical results were evaluated according to Lysholm and Lachman clinical rating scales.Results At the end of follow-up,there was no knee extension limitation and the knee flexion was between 110°and 150°,with an average of 130°.The Lysholm score of the 110 cases was (69.3±4.8) before operation, and (88.3±11.6) at the end of follow-up.The difference between the Lysholm scores was significant (P < 0.01), the difference of Lachman score below 4mm in 82 cases and 19 cases were below 5mm.Conclusion Early system rehabilitation training can relieve the postoperative pain and swelling of pain,and improve the function of knee joint in cruciate ligament injury of the knee joint patients.

[Objective] To construct a database for the genetic polymorphism of 18 STR loci (D8S1179, D21S11, D7S820, CSFIPO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, VWA, TPOX, D18S51, D5S818, FGA, PentaE, PentaD, SE33) in Hart population from Zhejiang province. To investigate the application of 18 STR loci in the field of paternity testing and prenatal diagnosis. [Methods] Fluorescent dye labeling multiplex STR-PCR, capillary electrophoresis and DNA sequencer GeneScan were adopted in genotyping 598 unrelated samples collected from Han population in Zhejiang province. 18-STR database was established and analyzed. Population comparison was conducted between Han population in Zhejiang province and 8 other population. 15-STR and 18-STR identification system were compared in 497 paternity testing cases. [Results] We observed the distribution of 18 STR loci in Han population meet Hardy-Weinberge equilibrium and was different from other 8 population (X~2 test, P>0.05). Statistical results showed that the heterozygosis (He) ranged from 0.630 to 0.942. The combined power of discrimination was>0.9999999999. Compared with 15-STR identification system, higher paternity index scores and higher exclusion rate were obtained with 18-STR identification system in dual-case paternity test and mutation identification. One trisomy 21 fetus was found in a prenatal paternity test case which had two characteristic genotypes in 2 STR loci of D21S11 and Penta D. [Conclusions] The 18 loci were relatively highly genetic polymorphic in Zhejiang Han population and could be used for paternity testing. Some STR loci could be used in prenatal diagnosis for aneuploidy.

Objective:To evaluate the efficacy of individualized cognitive behavioral therapy for unprotected sex and sexual attitude of middle school students having unprotected sex.Methods:A target sample of 68 adolescents having unprotected sex was recruited from 4 secondary schools in Changsha,Hunan [the unprotected sex (US) score of Health-Risk Behavior Inventory for Chinese Adolescents (HBICA) ≥ 1].Subjects were randomized assigned to cognitive behavioral therapy group (CBT group) and control group.Each group had 34 subjects.The CBT group was giving one-on-one counseling for 6 weeks (50 to 60 minutes weekly).The control group didn't receive intervention by counselors.The US and Attitudes Toward Sexuality Scale (ATSS) were selected as criterion measurements.Outcome assessments were made at baseline and at 1-and 3-month follow-up.Results:The reduction rate of US scores showed that the response rate of therapy was over 80％.Mixed linear model analysis showed that there were significant group effect,time effect and group × time effect in scores of US and ATSS (Ps ＜ 0.05).Simple effect analysis indicated that the scores of US and ATSS of CBT group were significant lower than those of baseline from 1-month follow-up [(2.2 ± 2.9) vs.(4.7 ± 3.1),(3.2 ± 1.6) vs.(4.7 ± 3.1);(38.2 ± 4.9) vs.(40.9 ±5.1),(37.2 ±5.4) vs.(40.9 ±5.1),Ps ＜0.01],whereas the scores of those in the control group did not show any significant difference (Ps ＞0.05).At l-month and 3 month follow-up,moderate effect sizes were found for the CBT and control groups on all the outcome measures (Cohen's d =0.50-0.70).Conclusion:The individualized cognitive behavioral therapy could effectively reduce the level of unprotected sex and sexual attitude of adolescents having unprotected sex.

Currently, the basic network resource of the entire traditional Chinese medicine (TCM) industry is fragmented. Issues of data unify are not easily resolved. It seems that there is no new breakthrough in key technologies, especially on the problem of data sharing and resources' integration. In this context, a method of medical information data sharing is advocated to solve the problem. We use the RESTful style to define resources of the pharmaceutical industry, which is based on the UDDI Web services standards. The objective of this paper is to create a method of TCM resource URL defined and exchanged. Finally, this paper provided an easy way for TCM data network sharing.

Objective To establish an HPLC method for the determination of ginsenosides Rg1 and Re in Shenqi Granula.Methods Chromasil C18 column(250 mm?4.6 mm)was used with acetonitrile-0.05% phosphoric acid solution(21∶79)as mobile phase.The flow rate was 1 mL/min and the detected wavelength was 203 nm.Results Ginsenosides Rg1 and Re could be baseline separated with in 30 min.The average recovery rates were 99.60% and 98.5%,corresponding RSD were 1.93% and 2.31% for ginsenoside Rg1 and Re,respectively(n=5).Conclusion This method is fast and accurate and can be used for quality control of Shenqi Granula.

0.05).CONCLUSIONS The rising rates of the drug-resistant Uu strains may be resulted of no-standard treatments.The treatment time will be shortened and the curative effects will be better if the treatments are based on the results of the susceptibility test.

Objective To explore the effect of melatonin on mitochondria in streptozotocin (STZ)-induced diabetic rats and its potential mechanism. Methods The diabetic rat models were induced by intraperitoneal injection of STZ, and the diabetic rats were randomly divided into two groups: diabetes mellitus group (DM group) and melatonin-treated group (DM± MT group). The normal non-diabetic rats were served as control group. Mitochondrial membrane potential and mitochondrial swelling were measured, the expression of voltage-dependent anion channel (VDAC)was assayed by immunohistochemistry in the heart, liver and kidney among the different groups after 8 weeks. Results (1)Compared with DM group, mitochondrial membrane potential in the heart,liver and kidney were significantly elevated in DM± MT group (553.6± 193.5 vs. 311.4 ± 133.7;745.7±115.8 vs. 358.9±158.7; 951.6±246.1 vs. 425.8±177.9, all P＜0.05). (2)Compared with DM group, mitochondrial swelling in the heart, liver and kidney was reinforced in DM ± MT group. (3)Compared with DM group, the expressions of VDAC in the heart, liver and kidney were significantly up-regulated in DM± MT group (76.93 ± 8.263 vs. 58.59 ± 7.62, 50.69 ± 6.33 vs.40.11±6.30, 77.86±8.59 vs. 61.44± 12.86, all P＜0.05). Conclusions Melatonin has protective effect on the activity of mitochondria in the heart, liver and kidney in diabetic rats possibly by up-regulating the expression of VDAC.