Objective To construct eukaryotic expression vector of attenuated Salmonella typhimurium containing hCTLA-4Ig cDNA and identify its expression in COS-7 cells for the further study of function in SLE models. Methods Touchdown PCR was used to amplify hCTLA-4Ig cDNA.The PCR product was ligated into the multiple clone site of eukaryotic expression vector pcDNA3.1(+) by gene recombination technique.Then the recombinated plasmid pcDNA3.1(+)-CTLA-4Ig was transfected into COS-7 cells using DOTAP.The expression of interest protein in the supernatant of the cell disruption was detected with SDS-PAGE and Western blotting.Results Restriction analysis and DNA sequence analysis showed that the CTLA-4Ig cDNA had been successfully inserted into pcDNA3.1(+) eukaryotic expression vector.The interest protein could be detected in the supernatant of cell disruption 48h after the transfection of pcDNA3.1(+)-CTLA-4Ig.This protein specifically bound with human CTLA-4 monoclonal antibody.Conclusion The eukaryotic expression vector containing hCTLA-4Ig gene was successfully constructed and bioactive interest protein could be successfully expressed in mammalian cells.

ObjectiveTo observe the clinical effect of Chinese medication plus warm needling in regulating the hypothalamo-pituitary-ovarian axis (HPOA) for anovulatory infertility.MethodThe relevant clinical data of sixty-seven women with infertility were comprehensively analyzed, to summarize the risking factors for anovulatory infertility, and then the subjects were randomized into an observation group and a control groupby their visiting sequence. The control group was intervened by orally taking Bushen Tiaojing Cupailuan (tonifying kidney, regulating menstruation, and promoting ovulation) decoction, while the observation group was by warm needling in addition to taking the same decoction. The clinical efficacies, growth of endometrium, and size of follicle were compared between the two groups.ResultWarm needling plus Bushen Tiaojing Cupailuan decoction produced a positive effect in treating anovulatory infertility, andthe pregnancy rate of the observation group was significantly higher than that of the control group (P<0.05).ConclusionThe effect produced by warm needling plus Bushen Tiaojing Cupailuan decoction has a comparatively high clinical value.

Flood damage is one of the ordinary natural calamities. The areas of middle and down of the Yangtze River and its south are the endemic area of schistosomiasis. As the flood damage,it causes property loss,and harm public and people health severely even. This paper aims at the status of catastrophic flood damage again in the Yangtze River basin ,analyzes the impact of schistosomiasis transmission by it,and indicates that at present,the process of schistosomiasis control in China is marching from transmission control to transmission interruption and elimination,but the frequent flooding exacerbates Oncomelania hupen?sis snail breeding area and schistosomiasis source spreading,which may influence,in a certain extent,the realization of the goal of schistosomiasis control in China. We should timely assess and monitor the impact and risk of schistosomiasis transmission by flood damage,and give the early warning and adopt emergency handling measures in time. This paper also proposes the disas?ter prevention measures according to flood damage stages(the early,middle,later,and post?flood)and the occurrence and de?velopment of damage and endemic situation,so as to control the risk and damage of the disease transmission in a high limit,to consolidate the current achievements of schistosomiasis prevention and control and guarantee the realization of the goal of schisto?somiasis interruption and elimination in China on the schedule.

[Objective]To report the treatment and outcome of treatment of discoid lateral meniscal tear by arthroscopic surgery.[Method]During the period between Jan.1999 and Jun.2005,77 patients suffered from discoid lateral meniscal tear have been treated by arthroscopic surgery in Xijing Hospital. The standard arthroscopic portals (anteromedial and anterolateral)were used.Lateral menisci were totally removed in the 21 patients (29.2%) of total 77 cases,whereas lateral menisci of 13 patients (18.1%) were removed partially and meniscus-plasty was applied in 38 patients (52.8%).Thirty-nine (54.2%) of 77 patients were males,and 33 (45.8%) females.Their average age was 29.4?10.9 years (range 8~51 year).Five patients were lost in follow-up.The mean follow-up period of the rest 72 patients was 26.2 ?9.3 months (range 12~44 months).The patients were assessed pre-operationally and post-operationally by the Lysholm score.Moreover,the results of Lysholm score were analyzed by student-t test.[Result]All patients had progress in pain,swelling,range of motion and knee function. No complication,which included intraarticular infection,iatrogenic injury,fibroarthritis or loss of range of motion,was found.Lysholm score was 66.8?5.2 in average (range from 50 to 78) before operation,whereas it improved to 96.4?3.6 in average (range from 85 to 100) after operation.According to student-t analysis,very significant statistical difference was found between the pre-operative and post-operative results (t=34.7,P

Objective To observe the changes of peripheral blood T lymphocytes,IFN-?,TNF-? and IL-4 in rats infected by T.gondii.Methods 48 Sprague-Dawley(SD)rats were intra-abdominally injected with 2?105/L of cellulose purified living tachyzoites in 2 ml and randomly divided into 8 groups.Six rat was intra-abdominally injected 2 ml of saline as control and 4 rats were remained as normal control.Peripheral blood was collected and the level of IFN-?,TNF-?,IL-4 was analyzed by ELISA on day 1,3,7,14,28,35,42,60.Results Level of IFN-?(6.73 pg/ml)and IL-4(6.91 pg/ml)increased in experimental rats on day 7(P

BACKGROUND:Animal models of osteoporosis play an important role in the research of the pathogenesis, occurrence and development of osteoporosis, as wel as in the clinical diagnosis, prevention and treatment of osteoporosis. OBJECTIVE: To summarize and discuss the establishment and research ideas of osteoporosis models, explore the current situation and advance of osteoporosis models, compare the advantages and disadvantages of various methods, and provide evidence for clinical investigation. METHODS: A computer-based online search was conducted in SinoMed, VIP, Wanfang and PubMed databases by using the key words of “animal model, osteoporosis” from January 1969 to October 2015. The language was limited to both Chinese and English. Relevant articles were screened according to inclusion and exclusion criteria. The documents about the methods of osteoporosis model preparation, method improvement as wel as their advantage and disadvantage were summarized. RESULTS AND CONCLUSION:A total of 576 articles were included. Among them, articles published earlier, duplicated, and similarly were excluded, and 53 articles were finaly included. Various animal models of osteoporosis may only focus on the certain causes, certain stage, some of the main symptoms and some pathophysiological changes of disease. Accordingly, appropriate modeling methods and experimental animals should be selected based on research objective. Rat undergoing castration is the most commonly used model in the modeling of osteoporosis. Among drug methods for constructing osteoporosis model, glucocorticoids is the most commonly used one. Disuse method and nutritional method have limitations, and always combined with castration and drug methods. The effects of gene transfer, gene mutation and brain-derived model deserve

Objective To explore the diagnostic value of the concentration of plasma circulation microRNA155 (miRNA155) in ulcerative cilitis (UC) and its correlation with clinical characteristic of UC.Methods From October 2010 to August 2012,a total of 136 patients diagnosed as UC were enrolled,and at same time,170 healthy individuals were set as healthy control.The blood samples of all participants were obtained and plasma was isolated.The adsorption column was used for RNA extraction according to miRNeasy kit instruction.RNA was reverse-transcribed into cDNA with miScript reverse transcription kit.cDNA was a template and miRNA155 real-time quantitative polymerase chain reaction (PCR) was performed with miScript SYBR Green PCR kit.The relative quantity of miRNA155 expression was calculated with 2-△△Ct method.Analysis of variance were performed for comparison between groups.Receiver operating characteristic curve analysis was used for the diagnostic value of miRNA155 concentration in UC.Multiple linear regression analysis was used for the correlation between miRNA155 concentration and clinical characteristics of UC.Results The concentration of plasma circulation miRNA155 of patients with UC ((1357.43±326.15) fmol/L)was higher than that of healthy controls ((1140.70 ± 312.47) fmol/L) and the differences were statistically significant (F=35.56,P＜0.01).The area under receiver operating characteristic curve of the concentration of plasma circulation miRNA155 of patients with UC was 0.847,and the 95 ％CI was 0.806 to 0.888 (P＜0.01).When the concentration of plasma circulation miRNA155 was 1404.51 fmol/L,its specificity in the diagnosis of UC was 94.7％,and sensitivity was 40.4％.There was correlation between the concentration of plasma circulation miRNA155 and the disease activity in patients with UC (F=12.91,P＜0.05).However there was no correlation with the severity and location of the disease (both P＞0.05).Conclusion Plasma circulation miRNA155 highly expressed in patients with UC,and its concentration is correlated with the disease activity.

Objective To approach the effect of different vasopressor on hemodynamics, volume responsiveness, fluid volume balance, renal function and prognosis in patients with acute respiratory distress syndrome (ARDS) complicated with septic shock.Methods A prospective single-blind randomized controlled trial was conducted. ARDS patients with septic shock admitted to the Department of Critical Care Medicine of Jiangxi Provincial People's Hospital from January 1st, 2015 to May 1st, 2016 were enrolled. The patients satisfied ARDS Berlin diagnostic criteria, over 15 years old, needing vasopressor after fluid resuscitation were enrolled. The patients were divided into norepinephrine group (NE group) and terlipressin group (TP group) by randomise number table derived by computer. Patients in TP group were given terlipressin (0.01-0.04 U/min) with an intravenous pump, while those of NE group were given norepinephrine (> 1μg/min) with an intravenous pump, and the target mean arterial pressure (MAP) was maintained at 65-75 mmHg (1 mmHg = 0.133 kPa). Hemodynamics and extravascular lung water index (EVLWI) were monitored by pulse indicator continuous cardiac output (PiCCO). The volume responsiveness of patient was evaluated by passive leg raising (PLR) test, and cardiac index (CI) change (ΔCI ≥ 10%) served as positive volume responsiveness. The differences in hemodynamics, EVLWI, oxygenation index (OI), lactate clearance rate (LCR), rate of positive volume responsiveness, urinary output, fluid volume balance, renal function, and prognostic indicators were compared between the two groups.Results Fifty-seven patients with ARDS complicated with septic shock were enrolled, with 26 patients in NE group, and 31 patients in TP group, thebaseline data in both groups was balanced with comparability. Compare with NE group, 48-hour and 72-hour heart rate (HR) in TP group was significantly slowed (bpm: 82.1±6.8 vs. 87.6±7.4, 81.3±6.1 vs. 85.6±8.3, bothP < 0.05), 72-hour central venous pressure (CVP) was significantly decreased (mmHg: 9.4±2.6 vs. 10.9±3.0,P < 0.05), but no significant difference was found in HR, MAP, CVP, CI, EVLWI, OI and LCR at other time points between the two groups. 48-hour and 72-hour positive volume responsiveness rate in TP group were significantly increased as compared with those of NE group (74.2% vs. 46.2%, 64.5% vs. 38.5%, both P < 0.05), urinary output on the 2nd day (mL/24 h: 2342.8±704.1 vs. 1944.6±684.3) and fluid volume balance (mL: -319.7±54.8 vs. -169.6±27.2) were significantly decreased (bothP < 0.05). There was no significant difference in positive volume responsiveness rate, urine output, fluid volume balance, and the level of serum creatinine at other time points between the two groups. There was no statistically significant difference in the following features between TP group and NE group: duration of mechanical ventilation (days: 8.41±2.97 vs. 9.67±3.56), length of intensive care unit (ICU) stay (days: 12.84±4.47 vs. 14.77±5.01), total length of hospital stay (days: 19.34±7.37 vs. 21.07±8.41), and 28-day mortality (29.0% vs. 30.8%, allP > 0.05).Conclusions Compared with norepinephrine, terlipressin for ARDS patients with septic shock is more conducive to restrict fluid load, improve the renal perfusion and increase urine output. However, in both groups therewas no significant difference in the efficiency of stabilizing hemodynamics, shortening the duration of mechanical ventilation, reducing ICU or hospital days and decreasing 28-day mortality.

This study aimed at verifying a previous patented fraud detection method of propolis.In accordance with the patented process,the chromatographic separation was achieved on a Kromasil 100-5C18 column (250 mm × 4.6 mm,5 tm) as mobile phrase A was methanol and mobile phrase B was water (0.5％ phosphoric acid) at the flow rate of 1 mL· min-1 for gradient elution.The detection wave length was 296 nm.Fraudpropolin A was taken as the reference,while the known sources of natural propolis were determined by HPLC.As a result,no fraudpropolin A was detected in the 134 sources of natural propolis at different types or from various origins.It was concluded that the patented process was sound in the fraud detection method of propolis.

Objective To study the expressions of metalloproteinase 2(MMP-2) and tissue inhibitors of metalloproteinase 2(TIMP-2) in lung cancer,and their relationships with microvessel density(MVD).(Methods The) expressions of MMP-2,TIMP-2 and Ⅷ factor were tested by immunohistochemical staining in(90 cases) of lung cancer tissue and 40 cases of para-cancer lung tissue.Results The immunohistochenmical staining results analyzed by IPP software indicated that MMP-2 and TIMP-2 expressed in all pathological types of lung cancer.The expreesion level of MMP-2 in lung cancer tissue was higher than that in para-cancer tissue(P