Objective To develop mycobacterial inducible expression vectors which permit to overexpress Mycobacterium tuberculosis (Mtb) immunodominant antigen, and to analyze its immunogenicity after purification by affinity chromatography. Methods The regulatory region of M. smegmatis (Ms) acet-amidase(pACE) was obtained by PCR amplification, and was used as promoter to construct the mycobacteri-al inducible expression vectors, pMF series. The coding gene of Mtb chimeric antigen Ag856A2 which is a recombinant Ag85A with 2 copies of ESAT-6 inserted in its Acc Ⅰ site and showed excellent immunogenicity in the animal experiments we described previously, was cloned into the pMF vector series, and was induced to express by the addition of acetamide. The recombinant protein expressed in the Ms was purified by the Ni~(2+)-NTA affinity chromatography, the resulted homologous recombinant antigen was added into the spleen cells separated from BCG vaccinated mice, and the immunogenicity was analyzed by the IFN-γ ELISPOT as-say. Results The mycobacterial inducible expression vectors, pMF series was constructed successfully, target antigen could be. induced to express in the Ms by the addition of 0.02% acetamide, and could be puri-fied by the Ni~(2+) -NTA affinity chromatography due to the addition of 6×His tag in the vector pMF406. Fur-thermore, the mycobactefial homologous antigen could induce more IFN-γ secretion than the heterogonous one. Conclusion The mycobacterial inducible expression system based on the regulatory region of Ms acet-amidase as promoter could permit the Mtb target antigen of interest overexpression and purification, and the immunogenicity of the homologous antigen from Ms is better than that of be expressed from E. coli, which may be more potential for immunological detection of tuberculosis.

BACKGROUNDTo screen and identify differentially expressed genes among lung cancer tissues, paracancerous pulmonary tissues and some other kinds of tumor tissues using suppression subtractive hybridization (SSH) and cDNA Microarray.

METHODSOne cDNA chip was made by gathering clones of three differentially expressed cDNA libraries which came from BEP2D cell lines during three different malignant transformed phases. Then the clones were hybridizated with cDNA probes which extracted from 15 cases of lung cancer tissues, 5 cases of paracancerous pulmonary tissues and 24 cases of other 8 kinds of tumor tissues respectively.

RESULTSTwenty-six cDNAs were obtained which expressed higher in lung cancer tissues than that in paracancerous pulmonary tissues. Thirty-one cDNAs expressed remarkably higher in paracancerous tissues than those in cancer tissues. Compared with other 8 kinds of tumors, paracancerous tissues had 63 overexpressed cDNAs and lung cancer tissues had 87 overexpressed cDNAs.

CONCLUSIONSThe combination of SSH and cDNA microarray is rapid and effective for screening and identification of differentially expressed genes in different samples. It may be potentially useful for diagnosis of lung cancer to further study the differentially expressed genes among lung cancer tissues, paracancerous pulmonary tissues and other tumor tissues.

OBJECTIVE:There are many kinds of biological agents for the treatment of rheumatoid arthritis in clinic,but the differences in therapeutic efficacy and safety are still unclear.The purpose of this study is to compare the differences in effectiveness and safety of different biological agents for the treatment of rheumatoid arthritis. METHODS:CNKI,VIP,WanFang,China Biomedical Literature System,PubMed,Cochrane Library,Web of Science,and Embase databases were searched to collect the randomized controlled trials on biological agents for rheumatoid arthritis that meet the requirements from inception to October 1,2022.The literature was selected by EndNote software,and the quality of the included literature was evaluated by RevMan 5.3 software.The software Stata 14.2 was used for direct meta-analysis and network meta-analysis of ACR20(American College of Rheumatology 20%response),ACR50(American College of Rheumatology 50%response),ACR70(American College of Rheumatology 70%response),erythrocyte sedimentation rate,and adverse reactions. RESULTS:Totally 39 articles were included,including 5 low-risk articles,4 high-risk articles,and the remaining 30 articles contained unknown risk bias,with a total of 13 treatment measures.The results of network meta-analysis:(1)In ACR20,infliximab combined with methotrexate(OR=5.54,95%CI:1.33-23.01,P<0.05),abatacept+methotrexate tablets(OR=3.21,95%CI:1.13-9.10,P<0.05),and tocilizumab(OR=2.95,95%CI:1.61-5.44,P<0.05)were better than methotrexate tablets.The probabilistic ranking of ACR20 was:infliximab+methotrexate tablets>abatacept+methotrexate tablets>tocilizumab>certlizumab>etanercept+methotrexate tablets.(2)In the aspect of ACR50,etanercept combined with methotrexate tablets(OR=4.04,95%CI:2.13-7.66,P<0.05),infliximab combined with methotrexate tablets(OR=4.79,95%CI:1.19-19.26,P<0.05),and tocilizumab combined with methotrexate tablets(OR=3.54,95%CI:1.36-9.22,P<0.05)had better therapeutic effects than methotrexate tablets.The probabilistic ranking of ACR50 was:etanercept+methotrexate tablets>infliximab+methotrexate tablets>tocilizumab+methotrexate tablets>tocilizumab>certlizumab+methotrexate tablets.(3)In terms of ACR70,the therapeutic effects of infliximab combined with methotrexate tablets(OR=8.00,95%CI:2.31-27.69,P<0.05),etanercept combined with methotrexate tablets(OR=4.26,95%CI:2.51-7.21,P<0.05),and tocilizumab combined with methotrexate tablets(OR=3.51,95%CI:1.82-6.80,P<0.05)were better than methotrexate tablets.The probabilistic ranking of ACR70 was infliximab+methotrexate tablets>etanercept+methotrexate tablets>tocilizumab+methotrexate tablets>certlizumab>adalimumab+methotrexate tablets.(4)In erythrocyte sedimentation rate,etanercept combined with methotrexate tablets(SMD=-9.23,95%CI:-16.55 to-1.92,P<0.05)was better than etanercept and methotrexate tablets(SMD=14.59,95%CI:7.28-21.91,P<0.05).The probabilistic ranking of erythrocyte sedimentation rate was etanercept+methotrexate tablets>infliximab+methotrexate tablets>etanercept>adalimumab+methotrexate tablets>methotrexate tablets.(5)In terms of adverse reactions,placebo(OR=0.62,95%CI:0.39-0.99,P<0.05)was better than infliximab and certlizumab(OR=0.44,95%CI:0.25-0.78,P<0.05).The probabilistic ranking of adverse reactions was placebo>infliximab>etanercept+methotrexate tablets>certlizumab>etanercept. CONCLUSION:Based on evidence from 39 randomized controlled trials,infliximab combined with methotrexate tablets(highly recommended)can be the first choice in clinic,and etanercept combined with methotrexate tablets(highly recommended)can be the second choice in terms of good effectiveness and safety.

Objective:To investigate the risk factors for hemorrhagic transformation (HT) after intravenous thrombolysis (IVT) in patients with acute ischemic stroke (AIS), and the predictive value of Neutrophil to lymphocyte ratio (NLR).Methods:Consecutive patients with AIS received IVT in Zhengzhou People’s Hospital from January 2021 to December 2022 were retrospectively enrolled. HT was defined as no intracranial hemorrhage was found on the first imaging examination after admission, and new intracranial hemorrhage was found on the imaging examination 24 h after IVT or when symptoms worsened. sHT was defined as HT and the National Institutes of Health Stroke Scale (NIHSS) score increased by ≥4 compared to admission or required surgical treatment such as intubation and decompressive craniectomy. The baseline clinical and laboratory data of the patients were collected, and NLR, lymphocyte to monocyte ratio (LMR), and platelet to neutrophil ratio (PNR) were calculated. Multivariate logistic regression analysis was used to identify the independent predictors of HT and sHT, and receiver operating characteristic (ROC) curve was used to analyze the predictive value of NLR for HT and sHT after IVT. Results:A total of 196 patients were included (age 65.37±13.10 years, 124 males [63.3%]). The median baseline NIHSS score was 4 (interquartile range: 2-10). Twenty patients (10.2%) developed HT, and 12 (6.1%) developed sHT. Univariate analysis showed that there were statistically significant differences in age, baseline NIHSS score, creatinine, NLR, and stroke etiology type between the HT group and the non-HT group (all P<0.05); there were statistically significant differences in age, NLR, PNR, creatinine, baseline NIHSS score, and stroke etiological type between the sHT group and the non-sHT group (all P<0.05). Multivariate logistic regression analysis showed that NLR was an independent predictor of HT (odds ratio [ OR] 1.375, 95% confidence interval [ CI] 1.132-1.670; P=0.001) and sHT ( OR 1.647, 95% CI 1.177-2.304; P=0.004) after IVT. The ROC curve analysis showed that the area under the curve for predicting HT by NLR was 0.683 (95% CI 0.533-0.833; P=0.007), the optimal cutoff value was 5.78, the sensitivity and specificity were 55.0% and 84.1%, respectively. The area under the curve for predicting sHT by NLR was 0.784 (95% CI 0.720-0.839; P=0.001), the optimal cutoff value was 5.94, the sensitivity and specificity were 66.67% and 84.24%, respectively. Conclusions:A higher baseline NLR is associated with an increased risk of HT and sHT after IVT in patients with AIS, and can serve as a biomarker for predicting HT and sHT after IVT.

Dynamic surgical navigation system has been wildly used in implantology, the navigation surgical system provide preoperative trajectory planning. Moreover, the constant visualization of drilling trajectory during operation assist the operators by avoiding critical anatomic structures to achieve safer surgery. Our article focuses on the development and function of dynamic navigation system to evaluate the accuracy of dynamic surgical navigation system when used for regular implants and zygomatic implants placement. We aim to discuss the accuracy of different brand of dynamic surgical navigation systems for implants placement and to investigate the main reasons led the inaccurate outcome.