Objective To construct the recombinant plasmid of fusion protein containing respiratory syncytial virus (RSV) cytotoxicity T lymphocyte (CTL) epitope M2:81-95 and heat shock protein (HSP) 70L1, and to investigate its immunogenicity after prokaryotic expression. Methods HSP70L1 gone was cloned from SMMC7721 cells. The M2:81-95 gene fragment was amplified by polymerase chain reaction (PCR) method. Plasmid pET-HSP70L1-M2 : 81-95 (pET-HSP70L1-M2) was constructed, identified and transferred into E. coli BL21 (DE3). Expression of HSP70L1-M2 : 81-95(HSP70L1-M2) was induced by isopropy-β-D-thiogalaetosidc (IPTG). The expressed protein was purified by affinity chromatography and renatured by gradient dialysis. The BALB/c mice were immunized with this fusion protein. IgG antibodies and the subtypes were detected by enzyme-linked immunosorbent assay (ELISA), and CTL responses were determined by methyl thiazolyl tetrazolium (MTT). Results The recombinant plasmid pET-HSP70L1-M2 was successfully constructed. The fusion protein HSP70L1-M2 was expressed in E. coll. The purified protein induced strong RSV-and CTL epitope-specific CTL responses and high titer of protein specific lgG antibody 4.87±0.35. The subtypes were IgG1 (5.53±0.28) and lgG2a (4.40±0.21) and IgG1/ IgG2a ratio was balanced. The titers of lgG, IgG1 and IgG2a in PBS control group were 0.33±0.17, 0.51±0.21 and 0, respectively, which werc significantly lower than those in immunized group (t = 3.512, 3.681, 5.856; P<0.05). Conclusions The recombinant plasmid pET-HSP70L1-M2 is successfully constructed and the fusion protein is expressed and purified. HSP70L1-M2 induced strong RSV-and CTL epitope-specific CTL responses and mixed T helper cell (Th)1/Th2 response in BALB/c mice.

Objective: To investigate the regulation of respiratory syncytial virus CTL epitope fused with G protein antigen fragment G1 to the specific immunoresponses. Methods: The recombinant plasmid pET-DsbA-G1 or pET-DsbA-G1F/M2 was transferred into E.coli BL21(DE3) and the fusion protein DsbA-G1F/M2 or DsbA-G1 was expressed.The expressing product was induced and purified by affinity chromatography. The two proteins were used to immunized BALB/c mice i.p, respectively. Serum and spleen cells were collected regularly. RSV-specific CTL responses were measured by MTT, IgG and IgG1 and IgG2a antibodies by ELISA, neutralizing antibodies by plaque reduction assay. Results: The recombinant proteins were expressed successfully and the purity was over 90% after purified by affinity chromatography. The protein DsbA-G1F/M2 induced significant RSV-specific CTL responses, while DsbA-G1 without CTL epitope did not induce detctable CTL responses. Strong IgG antibody responses were elicited,indicated by both. IgG1 and IgG2a titers induced by DsbA-G1F/M2, while only IgG1 was induced by DsbA-G1.The ratio of IgG1/IgG2a was downregulated significantly. Both antigens induced high level of neutralizing antibodies, but the latter was a little lower. Conclusion: DsbA-G1F/M2, the fusion protein of a CTL epitope and G protein fragment G1 can induce both cellular immunity and humoral immunity. The activation of CTLs downregulates the ratio of IgG1/IgG2a.The more balanced immunoresponse is advantageous for improving safety of the candidate vaccine.

Objective To investigate the clinical effect of uterine artery embolization chemotherapy in the treatment of special site pregnancy .Methods 36 patients with special site pregnacy were randomly divided into the observation group and control group .The control group received uterine curettage after uterine artery embolization treatment,the observation group received bilateral uterine artery perfusion of methotrexate combined with uterine cu -rettage after embolization therapy .Time of-HCG returned to normal ,intraoperative blood loss ,hospitalization time and incidence rate of complications were compared between the two groups .Results After treatment,the time of -HCG returned to normal,intraoperative blood loss and hospitalization time in the observation group were (27.3 ±3.2)h, (120.5 ±18.3) mL,(85.6 ±8.4) h,which were significantly less than (76.4 ±8.2) h,(375.1 ±68.4) ml and (147.1 ±10.5)h in the control group (t=6.75,4.54,4.43,all P<0.05).The incidence rate of complication in the observation group was 16.8%,which was significantly lower than 50.1% in the control group (χ2 =4.49,P <0.05).Conclusion The uterine artery chemotherapy and embolization in the treatment of special parts of pregnancy can significantly shorten the hospitalization time and time of -HCG returned to normal ,reduce the intraoperative blood loss,the clinical effect is good ,it can be used as an effective method for the treatment of special site pregnancy .

AIM To develop a UPLC-MS/MS method for the simultaneous content determination of adenosine,salidroside,chlorogenic acid,calycosin-7-O-β-D-glycoside,specnuezhenide,calycosin,formononetin,quercetin,apigenin and astragaloside Ⅳ in Zhenqi Fuzheng Capsules (Ligustri lucidi Fructus and Astragali Radix).METHODS The analysis of methanol extract of this drug was performed on a 40 ℃ thermostatic Inertsutain C18 column (75 mm × 3.0 mm,2 μm),with the mobile phase comprising of methanol-acetonitrile-4 mmol/L ammonium acetate flowing at 0.5 mL/min in a gradient elution manner.RESULTS Ten constituents showed good linear relationships within their own ranges (r≥0.996 0),whose average recoveries were 95.1％-104.3％ with the RSDs of less than 4.20％.CONCLUSION This sensitive and specific method can be used for the rapid quality control of Zhenqi Fuzheng Capsules.

Objective To investigate the dynamic changes and prognosis of thyroid function in the patients with chronic hepatitis C (CHC)after antiviral treatment,and to clarify the influence of baseline factors in the changes of thyroid function.Methods 243 CHC patients with normal baseline thyroid function were enrolled. All patients were treated with IFN-alpha-2b(IFN-α2b)combined with ribavirin for 48 weeks.The thyroid function and serum HCV RNA level were assessed at 12,24,36,48,60 and 72 weeks.According to the changes in thyroid function after treatment,the patients were divided into continued normal,subclinical hypothyroidism,hypothyroidism and hyperthyroidism groups.The regularity of the changes of thyroid function of the patients in various groups were observed.Results Among 243 CHC patients,82(33.7%)patients had thyroid dysfunction.The prevalence of subclinical hypothyroidism,hypothyroidism and hyperthyroidism were 20.9%(51/243),5.3%(13/243)and 7.4%(18/243),respectively. At the end of 72 weeks,there were 32 (39.0%)patients suffering from subclinical hypothyroidism,12 (14.6%) patients with hypothyroidism and 7 (8.5%) patients with hyperthyroidism rehabilitated.6(7.3%)patients suffering from hypothyroidism turned to subclinical hypothyroidism,and 3(3.7%) patients suffering from hyperthyroidism turned to subclinical hypothyroidism.19(23.2%)patients had no significant change,they performed for continued subclinical hypothyroidism (1,1.2%),hypothyroidism (13,15.9%)and hyperthyroidism (5 , 6.1%).In addition, 3 (3.7%)patients with hyperthyroidism turned to hypothyroidism.An increased risk for hypothyroidism was found in female patients compared with males (P<0.05);the average age of the patients with hyperthyroidism was lower than those of the patients with hypothyroidism, subclinical hypothyroidism and continued normal (P<0.05);the baseline levels of GGT in the patients with hyperthyroidism and hypothyroidism were lower than those of the patients with subclinical hypothyroidism and continued normal(P<0.05).The ratio of the patients with HCV 2a to the patients with hypertyroidism was higher than those of the patients with hypothyroidism,subclinical hypothyroidism and continued normal(P<0.05).Conclusion Thyroid function in the CHC patients can be affected by antiviral treatment. Gender, age, liver function, genotype of HCV are influencing factors for the changes of thyroid function.

Objective To discuss the short-term curative effect and the safety of percutaneous vertebroplasty using high viscosity bone cement for the treatment of severe osteoporotic vertebral body compression fractures. Methods The clinical data of 100 patients with severe osteoporotic vertebral body compression fractures (compression degree>70%), who received percutaneous vertebroplasty by using high viscosity bone cement during the period from December 2010 to May 2013, were retrospectively analyzed. All the patients were followed up for at least one month. Both preoperative and postoperative visual analogue scale (VAS) and quality of life (QOL) scores, as well as the incidence of bone cement leakage, were recorded and the results were used to evaluate the curative effect and the safety of percutaneous vertebroplasty for severe osteoporotic vertebral body compression fractures. Results One week after the treatment, significant pain relief was obtained in 92 patients (92%), VAS scores decreased from preoperative (7.0 ±1.2) to postoperative (2.0±1.5), and QOL scores increased from preoperative (30±5.0) to postoperative (80±18.0);the differences were statistically significant (P<0.01). One month after the treatment, significant pain relief was seen in 91 patients (91%), VAS scores decreased from preoperative (7.0 ±1.2) to postoperative (1.5 ±1.0) and QOL scores increased from preoperative (30±5.0) to postoperative (80±15.0); the differences were statistically significant (P<0.01). No statistically significant differences in VAS scores and QOL scores existed between the data determined at one week after the treatment and the data determined at one month after the treatment (P>0.05). Leakage of bone cement was observed in 40 patients (40%), resulting no severe neurological symptoms; among the 40 patients, intervertebral disc leakage at above and below the vertebral body was detected in 28 patients (70%), vertebral anterior edge leakage was observed in 11 patients (27.5%) and vertebral posterior edge leakage was seen in one patient (2.5%). Conclusion For the treatment of severe osteoporotic vertebral body compression fractures, percutaneous vertebroplasty by using high viscosity bone cement is safe and effective.

Objective: To investigate the causes of disease among patients with liver disease hospitalized in Department of Infectious Diseases in our hospital and the changes in such causes within the past 20 years. Methods: A retrospective analysis was performed for the clinical data of 7570 patients who were admitted to our hospital from January 1995 to December 2015. The chi-square test was used for the statistical analysis of constituent ratio. Results: Of all 7570 patients with liver disease, 4930 (65.13%) had viral hepatitis, 332 (4.39%) had immune disease, 215 (2.84%) had drug-induced liver injury, 192 (2.54%) had fatty liver disease, 88 (1.16%) had schistosome-induced liver disease, 160 (2.11%) had inherited metabolic diseases, and 20 (0.13%) had vascular disease; 689 (9.1%) still had no clear cause of disease at discharge. The proportion of patients with viral hepatitis was 77.61% in the first 10 years and 59.19% in the last 10 years (P < 0.01). As for liver disease caused by hepatotropic virus, there were significant increases in the proportion of patients with hepatitis C or hepatitis E from the first to the last 10 years (hepatitis C: 2.24% vs 15.56%, P < 0.01; hepatitis E: 18.61% vs 23.07%, P < 0.05), while there were significant reductions in the proportion of patients with hepatitis B (68.14% vs 60.01%, P < 0.05) or hepatitis A (10.7% vs 1.36%, P < 0.05). The proportion of patients with immune diseases was 0.82% in the first 10 years and 6.08% in the last 10 years (P < 0.01). There were also certain changes in the proportion of patients with liver disease caused by other reasons. Conclusion There is a large proportion of patients with viral hepatitis among patients with liver disease hospitalized in Department of Infectious Diseases in a large general hospital, especially hepatitis B and E caused by hepatotropic virus. There is a certain change in the epidemiology of liver disease within the past 20 years, with a reduction in the proportion of patients with viral hepatitis and an increase in the proportion of patients with non-infectious liver diseases. There is a large proportion of patients with unknown causes of liver disease.