BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.

OBJECTIVE To compare commonly used antibiotics including cefotaxime,ceftazidime imipenem and meropenem inductivity to L1,L2 ?-lactamases from Stenotrophomonas maltophilia.METHODS RT-PCR method was used to determine the expression of L1,L2 ?-lactamases under the condition of different concentrations(1/4,1,4?MIC)of antimicrobial agents.Electrophoresis strip of L1,L2 ?-lactamases was quantified by software Image J.Minimal inhibitory concentrations(MICs)were detected by agar dilution method when different concentrations of antibiotics were as inductors.RESULTS Different concentrations of four antibiotics being used as inductors,electrophoresis strips of L1,L2 amplicons were not found in strains of blank control and those whose media had imipenem,meropenem or cefotaxime(4?MIC).The weaker electrophoresis strips were found in the isolates whose media had ceftazidime(1/4,1,4?MIC)or cefotaxime(1/4?MIC),but the strongest electrophoresis strips,the most quantities of expression and higher MICs were shown in the isolates whose media had cefotaxime(1?MIC)(P

Aim To study effects of metfomin(MF) on calmodulin(CaM) mRNA exprssion of the pitutary-gonad axis in rats.Methods The CaM mRNA expression of the pitutary-gonad axis in rats was measured with the real time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Results MF (270 mg?kg-1?d-1,ig,21 d) significantly inhibited the CaM mRNA expression in testes (P

OBJECTIVE To investigate the conditions and associated risk factors of nosocomial infections caused by Stenotrophomonas maltophilia.METHODS Thirty isolates of S.maltophilia causing nosocomial infections were collected and identified with API 20NE test strips.Minimal inhibitory concentration(MIC) of 14 antimicrobial agents against 30 isolates was determined by broth microdilution method.Case-control study and multivariate Logistic regression analysis were used for statistics to verify risk factors of infections caused by S.maltophilia.RESULTS Thirty isolates of S.maltophilia were highly resistant to imipenem,meropenem,cefotaxime, aztreonam and amikacin,but showed certain susceptibility to cefoperazone/sulbactam,piperacillin/tazobactam,trimethoprim-sulfamethoxazole and ticarcillin/clavulanic acid(96.7%,76.7%,73.3% and 60.0%,respectively).The independent risk factors leading to infections of S.maltophilia were mechanical ventilation(OR=7.629) and over 60 days of length of stay(OR=4.466).CONCLUSIONS S.maltophilia shows multiresistance to commonly used antimicrobial agents.The mechanical ventilation and over 60 days of length of stay are the independant risk factors for nosocomial infections caused by S.maltophilia.

Objective To detect the expressions of hTERT and MMP-7 mRNA in the peripheral blood of patients with colorectal cancer and their correlation,and discuss the clinical significance of peripheral blood micro-metastasis.Methods Total RNA was extracted from blood samples by TRIzol and was reverse transcribed into cDNA.The mRNA expressions of MMP-7 and hTERT were detected by real-time PCR.Results Positive mRNA expression of hTERT was found in 63 of 181 patients with colorectal cancer,while positive mRNA expression of MMP-7 was seen in 61 cases.The positive rates of hTERT and MMP-7 mRNA had no significant differences in the patients' sex and age (P ＞ 0.05).The positive rate of MMP-7 mRNA in the patients with poorly differentiated adenocarcinoma (43.9 ％) was higher than that in the patients with moderately and well differentiated adenocarcinoma (27.8 ％) (x2 =4.873,P ＜ 0.05).The positive rate of HTERT mRNA in the patients with stage Ⅲ-Ⅳ (42.5 ％) was higher than that in the patients with stage Ⅰ-Ⅱ (24.0 ％) (x2 =6.591,P ＜ 0.05).The positive rate of MMP-7 mRNA in the patients with stage Ⅲ-Ⅳ (39.6 ％) was higher than that in the patients with stage Ⅰ-1Ⅱ (25.3 ％) (x2 =4.014,P ＜ 0.05).The positive rate of hTERT mRNA in the patients with distant metastasis (53.3 ％) was higher than that in the patients without distant metastasis (28.7 ％) (x2 =9.059,P ＜ 0.05).The positive rate of MMP-7 mRNA in the patients with distant metastasis(46.7 ％) was higher than that in the patients without distant metastasis (29.4 ％) (x2 =4.506,P ＜ 0.05).The positive rate of hTERT mRNA in the patients with positive CEA expression (52.6 ％) was significantly higher than that in the patients with negative CEA expression (30.1％) (x2 =6.735,P ＜ 0.05).The positive rate of MMP-7mRNA in the patients with positive CEA expression (60.5 ％) was significantly higher than those in the patients with negative CEA expression (26.6 ％) (x2 =15.490,P ＜ 0.05).In the patients without distant metastasis,any of hTERT and MMP-7 mRNA expression was thought to be blood micro-metastasis.The recurrence and metastasis rates after 6 months in the patients with blood micrometastasis (36.58 ％) were higher than those in the patients without blood micro-metastasis (7.14 ％)(x2 =13.027,P ＜ 0.05).The recurrence and metastasis rates after 12-month follow-up in the patients with blood micro-metastasis (68.29 ％) were higher than those in the patients without blood micro-metastasis (12.5 ％) (x2 =31.948,P ＜ 0.01).The sensitivity of combined detection was 80.00 ％,and the specificity was 79.03 ％ after 12-month follow-up.Spearman correlation analysis revealed that significant positive correlation existed between hTERT and MMP-7mRNA expression (r =0.341,P =0.000).Conclusion Detection of hTERT and MMP-7 mRNA expression can be used as reliable markers to predict peripheral blood micrometastasis in colorectal cancer patients.

Objective To observe the anesthetic effect of Sumianxin Ⅱcombined with chloral hydrate in plastic surgical operation on rabbit and explore the fast , safe and effective anesthetic methods .Methods A total of 82 Rabbits for operation were randomly divided into A , B and C groups.The A group was injected with SumianxinⅡintramuscularly (0.35 mL/kg by weight).The B group was injected with 10%chloral hydrate intravenously (2.5~3 mL/kg by weight).The C group was injected with Sumianxin Ⅱintramuscularly ( 0.2 mL/kg by weight ) combined with 10% chloral hydrate intravenously(1 ~1.5 mL/kg by weight).The anesthetic effect of induction and recovery time of anesthesia ,anesthesia maintaining time ,and anesthesia mortality rate were observed and compared .Results The anesthetic effect of induction of anesthesia,recovery time of C groups fall in between which of A and B groups ( P <0.05).However, as for anesthesia maintaining time , there was no significant difference between A and C group and both the two groups were longer than B group.Conclusion SumianxinⅡcombined with chloral hydrate possess the advantages of fast anesthesia induction and recovery effect , longer anesthesia maintaining time and low mortality .Therefore it is an easy to operate , expeditious , safe and effective anesthetization for plastic surgical operation on rabbit .

Objective To investigate the anti-inflammatory and analgesic effects of combined application of glucosamine(GS)and chondroitin sulfate(CS).Methods The acute and chronic anti-inflammatory effects of combined application of GS and CS were observed respectively through carrageen-induced paw edema and cotton-induced inguinal granuloma of rats.The analgesic effect of combined application of GS and CS was investigated by the mouse pain model induced by 0.6% acetic acid Results As compared with the control group,the degree of paw edema and the weight of granuloma in combined application of GS and CS group were significantly reduced(P<0.05 and P<0.01,respectively);and the writhing number of mice decreased significantly(P<0.05).Conclusion Combined application of glucosamine and chondroitin sulfate demonstrates obviously anti-inflammatory and analgesic effects.

Objective To observe the effectiveness of introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride .Method A total of 80 rabbits of both genders for operation were randomly divided into A , B and C groups .The A group was injected with Sumianxin intramuscularly ( 0.3 mL/kg by weight ) .The B group was injected with Sumianxin and ketamine hydrochloride intramuscularly ( 0.3 mL/kg by weight ) .The C group was injected with Diazepam intravenously ( 1.5 mL/kg by weight ) combined with Sumianxin and ketamine hydrochloride injected intramuscularly (0.3 mL/kg by weight).The aesthetic effects, induction time, anesthesia maintaining time, total anaesthetic dose and operation time were observed , recorded and compared .Result The induction time of the C group was significantly shorter than A and B groups (P<0.01).The initial anesthesia maintaining time of the C group was the longest among the three (P<0.01) with least total anaesthetic dose (P<0.01).The operation time of the C group was the least with best aesthetic effects (P<0.01).Conclusion Introducing Diazepam into combined aesthesia of Sumianxin and ketamine hydrochloride can improve the aesthetic effects .Therefore , this is an optional aesthetic method for time-consuming animal operation or sensitive surgical sites of rabbits .

Objective:To clone and express Mycobacterium tuberculosis(Mtb) Rv2460c gene(encoding ClpP2 protein),and evaluate the immunogenicity of its coding product. Methods: The recombinant plasmid of pET32a (+) vector-ClpP2 that H37Rv Rv2460c gene was cloned into the plasmid pET32a(+)vector,was transformed into E. coli BL21(DE3)and induced expression by IPTG,then purified by affinity chromatography. The recombinant protein was confirmed by SDS-PAGE and Western blot. The analysis of immunogenicity of Mtb ClpP2 and its epitope prediction were performed by bioinformatic methods. The antibody levels of polyclonal antibody titer against ClpP2 protein in rabbits and TB patients′ serum were detected by ELISA. Results: The recombinant ClpP2 protease was expressed as inclusion bodies in E. coli. The purity of purified protein was 93% by bandscan software analysis. The bioin-formatics analysis shows Mtb ClpP2 protein has multiple preponderant B cell and T cell epitopes. Rabbit antiserum titer was 1∶64 000;Serum anti-ClpP2 antibody levels in TB patients was higher than that in healthy control subjects. Conclusion:The recombinant ClpP2 protein was purified, and specific Rabbit anti-ClpP2 polyclonal antibody was prepared successfully. Experiment and bioinformatic information studies showed that Mtb ClpP2 protease has strong immunogenicity.

Objective To discuss the application value of transthoracic echocardiography (TTE) and lower extremity deep venous ultrasonography in patients of pulmonary thromboembolism (PTE) combined with coronary heart disease (CHD).Methods Nine hundred and seventy-four hospitalized patients of PTE in the first hospital of China medical university between December 2010 and March 2015 was enrolled,119 cases of these were combined with CHD (12.22％),including 94 cases of unstable angina and 25 cases of myocardial infarction.To review the general information,embolism area and ultrasonic report of the patients of PTE combined with CHD.The patients were divided into two groups according to whether they were combined with left ventricular systolic or diastolic dysfunction or valvular diseases.To compare TTE parameters between the two groups with two-sample t-test and calculate the sensitivity and specificity of diagnosing imaging massive pulmonary embolism (MPE) by TTE.Results (1) Seventeen cases thrombosis (3.43％) were observed in PTE patients who underwent TTE.The majority were pulmonary artery thrombosis (11 cases).Four cases of thrombosis (5.63％) were observed in PTE patients combined with CHD.The majority were right heart embolus (3 cases).(2) Lower extremity deep venous thrombosis (LEDVT)was observed in 52.60％ of all the PTE patients and 34.62％ of PTE patients combined with CHD.The majority were on the left and the deep venous thrombosis of the calf.(3) Fouty-six cases (64.79％) were PTEindirect signs in TTE.The sensitivity of echocardiography diagnosis of MPE was 84.38％ and the specificity was 52.60％.(4) Pulmonary arterial systolic pressure [(47.90± 21.49) mmHg vs (31.18± 22.43) mmHg(1 mmHg=0.133 kPa),t=3.227,P ＜0.05] and inferior caval vein diameter [(19.66±4.41) mm vs (16.20± 3.46) mm,t=3.598,P ＜ 0.05] in CHD combined with left ventricular systolic or diastolic dysfunction or valvular heart disease were higher than the control group.The differences were statistically significant.Conclusions The proportion of CHD in hospitalized PTE patients was high.TTE could find PTE signs and had high sensitivity for diagnosis ofMPE.TTE could prompt the possibility of PTE combined with left heart disease.