Objective To analyze the composition of the pharmacodynamic components in the activated prothrombin complex concentrate(aPCC) developed by our research group,and to study the relationship between each pharmacodynamic component and the coagulation factor Ⅷ(FⅧ) bypassing activity.Methods The self-made aPCC concentrates were used as the research object,and the FⅧ bypassing activity was firstly detected by the coagulation method,and then the activities of a series of non-activated coagulation factors,activated coagulation factors and anti-coagulation factors were detected by the coagulation method and chromogenic substrate method.The prothrombin complex concentrate(PCC) in the market was used as a control to analyze the correlation between each pharmacodynamic component and FⅧ bypassing activity.Results The FⅧ bypassing activity of self-made aPCC concentrates and control PCC products were(43.53±3.07) IU/mL and(0.10 ±0.02) IU/mL,respectively,with a significant difference(t=20.16,P <0.01).Compared with the control PCC products,in the non-activated coagulation factors of self-made aPCC concentrates,the activities of coagulation factor Ⅶ(FⅦ) and coagulation factor Ⅸ(FⅨ) were significantly different(t=22.72 and 8.00,respectively,each P <0.05),and the activities of coagulation factor Ⅱ(FⅡ) and coagulation factor Ⅹ(F Ⅹ) showed no significant difference(t=1.67 and-0.96,respectively,each P> 0.05);in the activated coagulation factors of self-made aPCC concentrates,the activities of activated coagulation factor Ⅱ(FⅡ a),activated coagulation factor Ⅸ(FⅨ a),and activated coagulation factor Ⅹ(F Ⅹ a) were significantly different(t=15.92,32.93 and 34.64,respectively,each P <0.01),and there was no significant difference in the activity of activated coagulation factor Ⅶ(FⅦa)(t=2.34,P> 0.05);in the anti-coagulation factors of self-made aPCC concentrates,there was a significant difference in protein S(PS) activity(t=12.82,P <0.01),and there was no significant difference in heparin content or protein C(PC) activity(t=0.85 and-0.34,respectively,each P> 0.05).The FⅧ bypassing activity in the self-made aPCC concentrates and PCC products was significantly correlated with the activities of F Ⅶ,FⅨ,F Ⅱ a,FⅨ a,FⅩa and PS(r=0.999,0.971,0.988,0.994,0.974,and-0.984,respectively,each P <0.01),and was correlated with F Ⅹ a/F Ⅱ(r=0.827,P <0.05).Conclusion Self-made aPCC concentrates contain higher FⅦ bypassing activity,which may be related to the high activity of F Ⅶ,FⅨ,F Ⅱ a,FIX a and FⅩa.

AIM: To establish a HPLC for the determination of rubimaillin in Shenqiangujing Granules (Radix Codonopsis, Radix Rubiae, Radix Rehmanniae, Fructus Ligustri Lucidi, etc.) METHODS: HPLC column was C-18 (250 mm?4.6 mm, 5 ?m). The mobile phase was methanol-water-tetrahydrofuran (310∶90∶3). Flow rate was 1.0 mL?min -1 . The detection wavelength was at 249 nm. RESULTS: The linear range of rubinaillin was in the range of 0.056～0.29 ?g, r=0.9999. The average recovery was 99.36% (RSD=1.60%, n=5). CONCLUSION: The method is accurate with the good reproducibility, and can be used for the determination of rubimaillin in Shenqiangujing Granules.

Dengue is the most common vector borne viral disease of humans globally.Detection of viral RNA from suspected patient specimens is rapid,specific and confirmative in laboratory diagnosis of dengue infections during the acute phase.In this study,a multiplex nested reverse transcription PCR (RT-PCR) system was established for clinical specimens.While other nucleic acid amplification tests showed relatively low sensitivity,the multiplex nested RT PCR assay detected 4 cases among blood clots from 8 serologically confirmed dengue patients.These results suggested that blood clots of dengue patients could be used in laboratory diagnosis,and that the multiplex nested RT PCR assay,which simplified the detection procedure,could facilitate viral RNA detection of specimens in clinical laboratories.

Objective To evaluate the effect of long-term glucocorticoid administration on nondepolarizing muscle relaxants-induced neuromuscular blockade in rat diaphragma in vitro.Methods Forty adult male SpragueDawley rats,aged 8 weeks,weighing 180-220 g,were randomized into 2 groups (n =20 each):control group (group C) and dexamethasone group (group D).In group D,dexamethasone 0.6 mg/kg was injected intraperitoneally once a day for 14 consecutive days,while the equal volume of normal saline was injected instead of dexamethasone in group C.The left phrenic nerve-hemidiaphragm was removed at 24 h after the last injection to evaluate the effect of d-tubocurarine.Different concentrations (0-10 μmol/L) of d-tubocurarine were added.The d-tubocurarine concentration-effect curve was drawn.The half inhibitory concentration and 95 ％ confidence interval of d-tubocurarine were calculated.Right diaphragma was removed for determination of the expression of embryonic nicotinic acetylcholine receptor (nAChRγ) mRNA and adult nicotinic acetylcholine receptor (nAChRε) mRNA by RTPCR.Results Compared with group C,the concentration-effect curve was shifted to the right,the half inhibitory concentration was significantly increased,and the expression of nAChRγ mRNA and nAChRε mRNA was up-regulated in group D (P ＜ 0.05).Conclusion Long-term glucocorticoid administration can weaken nondepolarizing muscle relaxants-induced neuromuscular blockade in rat diaphragma in vitro,and up-regulation of nAChR mRNA expression may be involved in the mechanism.

Objective:To understand the development reality of pastes in Hubei province through the investigation of paste busi-ness in 2013. Methods:Issued by Hubei provincial administration of traditional Chinese medicine, a questionnaire investigation was a-dopted in the study. The implementation, usage amount, scope, composition of the users, price, technical personnel and propaganda of pastes were investigated and statistically analyzed in 91 TCM, integrated Chinese and western and national hospitals in our province. Results:The business conduction rate of pastes in Hubei province was nearly 61. 54% in 2013 with the production of approximately 45,000 batches and the technical personnel of 1 538. The paste business created great economic value of 0. 48 million yuan in 2013 with strengthened propaganda. However, the brand building was slightly weak. Conclusion:The business of pastes in Hubei province has developed rapidly and still shows great development potential.

Since several dengue viruses (DENV) have been isolated in Fujian Province during the past decade, sequencing and evolution analyses of viral envelope genes are helpful in determining their possible transmission origins. In this study, viral RNA was extracted from 12 DENV strains from Fujian between 20042010. Viral envelope genes were amplified, cloned into TA vectors and sequenced, and the sequence data were subsequently analyzed by bioinformatics software. Full-length E genes of DENV-1 or DENV-2 of 1 485 bp, and DENV- 3 of 1 479 bp were obtained. It was indicated, from BLAST analysis and phylogenetic trees, that DENV strains in Fujian Province during 20042010 shared the highest similarity with Southeast Asian strains, suggesting that DENV circulating in Fujian Province between 20042010 were probably imported from Southeast Asia. Hence, extensive monitoring on passengers from this region at the entry-ports should be strengthened.

13 C isotopic abundance of intracellular free amino acid with a characteristic of fast- turnover can quickly reflect changes in intracellular metabolic state. But the concentration of intracellular free amino acid is low, the existed 13 C isotope detection method based on GC-MS can not satisfy the requirement with full scan mode. In this study, the selected ion monitoring method was used to detect accuracy higher likelihood of analysis of 13 C isotopic abundance of free intracellular amino acid. First, in the full scan mode we analyzed of the fracture law of different amino acids, found the feature corresponding to each amino acid fragments, and established 16 kinds of free intracellular amino acids characteristic fragment library. Then using this characteristic fragment library, only specific m/z signal was detected in sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards showed that the signal-to-noise ratio, measurement precision and accuracy were improved by 17, 2. 0 and 3. 8 times compared with the full scan mode. In the analysis of coenzyme Q10 producing strains of samples, this method was successfully used to detect isotopic abundance of 8 kinds of free intracellular amino acids. This method plays an important role in the detection of 13 C isotopic abundance of the intracellular free amino acid in cell metabolism research.

Objective:To establish an HPLC method to determine the entrapment efficiency (EE) and drug loading (DL) of curcumin (CUR)and quercetin (QUE)loaded self-microemulsifying drug delivery system.Methods:A centrifugation method was used to isolate the free drug.The content of drug was determined by HPLC.The analytical column was a Purospher STAR LP C18 column (250 mm×4.6 mm,5 μm) and the column temperature was 30 ℃.The mobile phase was acetonitrile-4% acetic acid (50∶50) and the flow rate was 1.0 ml·min-1.The UV detection wavelength was set at 370 nm and the injection volume was 10 μl.Results:CUR and QUE were linear within the range of 10.728-96.552 μg·ml-1 (r=0.999 8) and 1.08-9.72 μg·ml-1 (r=0.999 9),respectively.The average recovery was 99.98%(RSD=1.46%,n=9) and 100.34%(RSD=1.06%,n=9),respectively.In CUR-QUE-SMEDDS,the EE of curcumin and quercetin was (95.97±0.50)% and (95.91±2.52)%,and the DL was (25.82±0.15) mg·g-1 and (1.80±0.05)mg·g-1,respectively.Conclusion:The method is accurate,rapid and simple,and suitable for the determination of DL and EE in CUR-QUE-SMEDDS.

Objective To evaluate the association of the size of thyroid nodules and the diagnostic value of repeat fine needle aspiration cytology(FNAC) in diagnose of thyroid nodules.Methods One hundred and forty-three thyroid nodules in one hundred and thirty-five patients with repeat FNAC and thyroidectomy were retrospectively analyzed.The nodules were divided into three groups according to the largest diameter(L):GroupA,L＜0.5 cm;GroupB,0.5 cm≤L≤1.0cmandGroupC,L＞1.0cm.Yield of FNACs were divided into six levels according to the classification criteria of the Bethesda system,and Level Ⅰ,Ⅲ,Ⅳ,Ⅴ were defined as the indefinite cytological diagnosis.Results The diagnostic rate of repeat FNAC was obviously higher in group B compared with group A and group C,with significantly statistical difference (P ＜0.05).The risk of malignancy for persistently indefinite diagnostic aspirates after repeat FNAC was significantly reduced with statistical difference from the initial indefinite diagnostic aspirates in group B (P ＜0.05),while there was no statistical difference between group A and group C (P＞0.05).The diagnostic accuracy of repeat FNAC in group B was significantly higher than that in group A(P ＜0.05),but not in group C (P ＜0.05).Conclusions The repeat FNAC has a higher efficiency in those nodules with a largest diameter between 0.5 cm and 1.0 cm,and a repeat FNAC can be suggested.

Background and purpose:Due to the lack of cost-effective pre-treatment predictors for advanced cervical squamous cell carcinomas treated with concurrent chemoradiotherapy (CCRT), both baseline circulating CD4+CD25+CD127Low/- regulatory T cell (Treg) count and serum squamous cell carcinoma antigen (SCC-Ag) level were measured for this feasibility study. Methods: Peripheral blood samples were collected from 44 patients with stageⅡB-ⅣA cervical squamous carcinomas before CCRT. Flow cytometry immunophenotyping and enzyme-linked immunosorbent assay were used for circulating CD4+CD25+CD127Low/-Treg count and serum SCC-Ag level testing,respectively. Clinical and pathological characteristics were retrospectively reviewed to analyze the predictive value of the 2 indexes. Results:The baseline circulating CD4+CD25+CD127Low/-Treg count was lower in the patient group with positive treatment response than in the group with negative response [(8.78±2.80)%vs (10.95±2.56)%, P<0.05], and the serum SCC-Ag level showed no signiifcant difference between the 2 groups. No correlation was detected between these 2 markers (Spearman’rho=-0.093, P=0.540). Determined by plotting receiver operating characteristic curves, the best cut-off points were 9.76%for circulating CD4+CD25+CD127Low/-Treg count and 9.50 ng/mL for serum SCC-Ag level, respectively. Univariate analysis showed that pretherapeutic circulating CD4+CD25+CD127Low/-Treg count (OR=1.901, 95%CI:1.112-3.219, P=0.017), but not serum SCC-Ag level (OR=0.998, 95%CI:0.001-4.253, P=0.897), was predictive of clinical response to CCRT. Multivariate Logistic regression analysis revealed that pre-treatment CD4+CD25+CD127Low/-Treg count was an independent predictor for clinical response to CCRT (OR=3.115, 95%CI:1.253-7.742, P=0.014). Conclusion:Pretherapeutic circulating CD4+CD25+CD127Low/-Treg count is a feasible method to predict clinical response to CCRT in patients with advanced cervical squamous cell carcinomas.