Objective To compare the effect of three methods in diagnosis of plague by detecting of Yersina pestis F1 antigen. Methods In natural foci of plague, wild animal samples, such as blood, liver, spleen,and lymphoid tissue were collected, and the three methods of enzyme linked immunosorbent assay (ELISA),reverse indirect hemagglutination assay(RIHA) and gold-immunochromatography assay(GICA) were employed to detect F1 antigen of Yersina pestis. Results Total of 414 infused organ samples of natural death and captured wild animals in natural foci of plague were determined. Positive samples detected by GICA and ELISA were the same,the positive rates were 5.31%(22/414), both positive and negative coincidence rates were consistently 100%. Only 18 samples were positive by retrial in 186 samples with more than 2 holes aggregation by preliminary examination of RIHA, with nonspecific agglutination rate of 40.6% (168/414) and positive rate of 4.35% (18/414). The positive coincidence rate was 81.82% (18/22) between RIHA with GICA and ELISA, and negative coincidence rate was statistically significant(t = 4.379, P ＜ 0.01). Conclusions ELISA, RIHA and GICA can be used for early diagnosis of plague by detecting F1 antigen. The results of RIHA have quantitative significance, with higher non-specific agglutination rate, and heavy workload of re-examination; GICA and ELISA has the same specificity and sensitivity, but the results of GICA is only qualitative. ELISA excluded the defect of RIHA and GICA, and combines the advantages of both methods.

Objective To analyse the feasibility of detecting F1 antibody to Yersinia pestis in flushing fluid of heart blood of Rhombomys opimus with enzyme linked immunosorbent assay(ELISA) method and its application value in surveillance of the disease. Methods Serum, flushing fluid of heart blood and infusion fluid of liver and spleen of Rhombomys opimus, which were caught by capture in the plague focus of Zunger basin in 2007, were taken to carry out detection for F1 antibodies to Yersinia pestis with ELISA method. The data were processed with SPSS 17.0. Results Positive rate and average titer of serum were 12.35%(11/162) and 25.35, of flushing fluid of heart blood were 10.49%(17/162) and 23.75 and of the infusion fluid of liver and spleen 6.79%(17/162) and 2240,respectively. No statistical difference was found in positive detection rate when it was compared between serum and flushing fluid of heart blood(χ2 = 1.333, P ＞ 0.05), but it was obviously different between serum and infusion fluid of liver and spleen(χ2 = 7.111, P ＜ 0.01 ) and between flushing fluid of heart blood and infusion fluid of liver and spleen(x2 = 6.250, P ＜ 0.05). There was a significant difference in average titer between serum, flushing fluid of heart blood and infusion fluid of liver and spleen(t = 2.290, 3.612, P ＜ 0.05 or ＜ 0.01 ). The plague F1 antibody positive coincidence rate of serum and flushing fluid of heart blood was 85.0%(17/20), of serum and infusion fluid of liver and spleen was 55.0% (11/20), and of flushing fluid of heart blood and infusion fluid of liver and spleen was 64.7%(11/17). Conclusions The ELISA method can detect Fl antibody in flushing fluid of heart blood,and the method is feasible in plague surveillance.

AIM To investigate the protective effects of Dahuang Zhechong (DHZC) Pills (Rhei Radix et Rhizoma,Eupolyphaga seu Steleophaga,Hirudo,etc.) against alcoholic liver fibrosis (ALF) injury in mice and to explore the underlying mechanisms.METHODS C57BL/6 male mice were used to build up ALF injury model,intervened with DHZC Pills.The serum of mice was examined for changes in alanine transaminase (ALT),aspartate aminotransferase (AST),interleukin-6 (IL-6),interleukin-10 (IL-10),interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α).Simultaneously,the deposit of collagen 1 (COL-1) and apoptotic cell death in liver tissues were analyzed by immunofluorescent and TUNEL assay,respectively.The expressions of cleaved caspase-3 (CC3) in livers were measured by Western blot.RESULTS Compared with the model group,the levels of serum ALT,AST,IL-6,IFN-γand TNF-α of mice in DHZC group were decreased significantly.And the level of serum IL-10 of mice in DHZC group was increased significantly.Mice in DHZC group had higher rates of COL-1 deposition and apoptotic cell death in liver tissues than those in the model group.Mice treated with DHZC Pills showed lower expression of CC3.CONCLUSION DHZC Pills confers protection against ALF injury in mice by inhibiting the generation of COL-1 and down-regulating apoptosis of liver cells death as a result of adjusting the levels of inflammatory factors.

Objective To compare the diagnostic value of renal ultrasound scan (RUS) and 99Tcmdimercaptosuccinic acid (DMSA) renal scintigraphy in children with acute pyelonephritis (APN). Methods In all, 165 children with initial clinical diagnosis of APN, aged from 1.5 months to 11 yrs ( median 20 months), were included in the study, all of which were examined with RUS and DMSA renal scientigraphy. The diagnosis with DMSA renal scientigraphy results was taken as the standard reference to evaluate the diagnostic sensitivity and specificity of RUS. Results Of 99 out of all 330 kidneys that were found abnormal on DMSA renal scientigraphy, 31 were abnormal on RUS. Of the rest normal kidneys on DMSA scans renal scientigraphy, 4 were abnormal on RUS. Thus diagnostic sensitivity of RUS for APN was 31.3%(31/99) and specificity was 98.3% (227/231). Conclusions Although RUS provides with high diagnostic specificity for children with APN, its low sensitivity may underestimate the clinical evaluation of APN.More often than not, 99Tcm-DMSA renal scientigraphy is a clinical necesscity for the definite RUS diagnosis.

Objective To evaluate the success rate,vascular patency time and their influencing factors of percutaneous transluminal angioplasty (PTA) in treating swollen hand syndrome in hemodialysis patients.Methods The clinical data of 16 hemodialysis patients with swollen hand syndrome,who were admitted to authors' hospital during the period from May 2015 to March 2017 to receive PTA,were retrospectively analyzed.The technical success rate,the follow-up primary vascular patency time and primary patency rate were calculated,and the factors influencing technical success rate and vascular patency time were analyzed.Results Venography with DSA revealed that a total of 16 segments of venous stenosis or occlusion were found in 16 patients,including 6 stenotic lesions and 10 occlusive lesions.Successful PTA was obtained in 14 patients,including one patient whose angiography performed immediately after PTA with balloon dilatation showed that the stenosis was still over 50％,and stent implantation had to be carried out.The technical success rate was 87.5％,in 2 patients PTA failed as the guide wire could not pass through the long segment of vascular occlusion.The 14 patients were followed up for 3-24 months,and the median patency time was 10.5 months.The 3-,6-and 12-month primary patency rates were 71.4％ (10/14),57.1％ (8/14) and 42.9％ (6/14) respectively.Univariate analysis indicated that the length of occlusive segment and the balloon pressure required for angioplasty were the potential factors that affected the postoperative vascular patency time.Conclusion For the treatment of swollen hand syndrome in hemodialysis patients,PTA is safe and effective,although long-term vascular patency rate needs to be further improved.

Background Bone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17β-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7. Methods After the treatment of MCF-7 cells with E2 at different concentrations (10-11 mol/L, 10-9 mol/L, 10-7 mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10-7 mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor α (Erα) on BMP-6 promoter in the presence of E2. Results E2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10-7 mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P<0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of Erα on 1/2 ERE response element in BMP-6 promoter was further validated. Conclusion Estrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor Erα binding on 1/2 ERE element in the BMP-6 promoter.

Objective To investigate the oncolytic effect of E1B mutant adenovirus (d11520) on human malignant gliomas. Methods Ad-βgal vector was used to investigate the infectibility of dl1520.U251,Hep3B (positive control) and T24 (negative control) cell lines were infected with dl1520respectively at 50,5,0.5,0.005 and 0 pfu of multiplicity of infection (MOI).The replication efficiency of d11520 in host cells was assessed by plaque assay.The cytopathic effect (CPE) was assessed by crystal violet staining in a panel of tumor cells. Results Crystal violet staining showed the Hep3B cell line was the most sensitive to dl1520 and had the fastest CPE,followed by the U251 cell line,while the T24cell line had no CEP.The replication and infection rates ofdl1520 in the U251 cell line were lower than in the Hep3B cell line but significantly higher than in the T24 cell line (P＜0.05). Conclusion The E1B mutant adenovirus (dl1520) has a significant oncolytic effect on human malignant gliomas.

Background Restylane, a hyaluronic acid gel, has been widely used as a dermal filler in USA and European countries. This study was designed to study the safety and efficacy of Restylane as a non-permanent dermal filler for facial augmentation therapy in China for the correction of nasolabial folds during a follow-up period of 6 months. Methods The study consisted of a screening visit, a baseline visit during which injection with Restylane was given, and follow-up visits after four weeks, three months and six months. The efficacy was subjectively assessed by comparing the treatment results between pre-treatment and post-treatment. Adverse events were analyzed by severity and duration. Results At six months post-baseline, the subjects and the investigators' independent assessment score of Wrinkle Severity Rating Scale was decreased. Global Aesthetic Improvement Scale was considered to have improved by over 90% in some cases. Adverse events related to product and injection was reported in 21.5% of the injections. A vast majority of the post-treatment reactions were mild. Conclusions The efficacy of Restylane for nasolabial fold in a Chinese population was excellent. Restylane was well tolerated and no systemic reactions or other safety concerns were raised.

Objective To investigate the role of antigen-processing machinery (APM) component defects in HLA class Ⅰ antigen down-regulation in laryngeal squamous cell carcinoma (SCC) and to assess the clinical significance of these defects. Methods Fifty-one formalin-fixed,paraffin-embedded SCC specimens were examined for the expressions of APM component transporter associated with antigen processing (TAP1) and low molecular weight polypeptide (LMP-7) and HLA class Ⅰ antigen by immunohistochemistry.Results HLA class Ⅰ antigens,TAP-1 and LMP-7 expressions were downregulated in 56.9％ (29/51),39.2 ％ (20/51) and 45.1％ (23/51) of the tested specimens respectively,whereas HLA class Ⅰ antigens,TAP-1 and LMP-7 expressions lost in 21.6 ％ ( 11/51 ),33.3％ ( 17/51 )and 27.5 ％ ( 14/51 ) of the tested specimens respectively.TAP-1 and LMP-7 expressions were significantly correlated with HLA class Ⅰ antigen expression (r =0.460,P < 0.05 and r =0.685,P < 0.05,respectively).HLA class Ⅰ antigens down-regulation was significantly correlated with T stage( x2 =8.61,P ＜ 0.05).Both TAP-1 and LMP-7 down-regulations were significantly correlated with T stage ( x2 valueswere 9.72 and 8.97 respectively,P ＜ 0.05 ) and TNM stage( x2 values were 9.18 and 7.70 respectively,P ＜0.05 ).TAP-1,LMP-7 and HLA class Ⅰ antigen down-regulations were significantly associated with reduced patients' overall survival ( P ＜ 0.05 ) and disease-free survival ( P ＜ 0.05 ).Multivariate analysis showed lymph node metastasis,recurrence and HLA class I antigen down-regulation were unfavorable prognostic factors(P ＜ 0.05).Conclusions Down-regulated expressions of HLA class Ⅰ antigen and APM component TAP-1 and LMP-7 occur frequently in laryngeal squamouss cell carcinoma,by which cancer cells could avoid immune surveillance,while HLA class Ⅰ antigen down-regulation is a major contributing factor to tumour progression and mortality.

Objective To establish a HPLC to analyze the content of quercetin and kaempfer in the Camptotheca acuminata Decne leaves.Methods The sample of Camptotheca acuminata Dene leaves were obtained by 5％ hydrochloric acid-methanol The content of quercetin and kaempferol was measured by HPLC with chromatographic column:Hubble C18 (4.6 mm× 250 mm,5 μm),mobile phase:CH3 OH-0.2％ H3PO4(30:70),flow rate:1 mL·min-1,column temperature:30 ℃,detection wavelength:370 nm.The specificity,standard curve and limit of quantification (LOQ),precision and recovery,stability and repeatability were measured.Results The quercetin and kaempferol existed a good linear relation at 5.53-69.08 μg · mL-1 and 3.99-49.90 μg · mL-1,respectively.The LOQ of quercetin and kaempferol was 4.23 and 7.81 ng,respectively.And the average spotting recovery rate of quercetin and kaempferol was 98.94％ and 98.29％,respectively.The RSD values of inter-day and intra-day assays were lower than 0.89％ and 1.04％ for quercetin and kaempferol,respectively.Conclusion This method is an accurate,reliable and convenient method,which is suitable for the quality control of the leaves of Camptotheca acuminata Decne.