Objective To express and purify Chlamydial protease-like activity factor(CPAF)from Chlamydophila pneumoniae,for investigating the effect of its recombinant protein GST-CPAF in inducing human monocytic cells to secrete proinflammatory cytokines and cell apoptosis.Methods The recom-bination expression plasmid pGEX6p-2/CPAF from Chlamydophila pneumoniae was transformed into E.coli.The recombination GST-CPAF was expressed after induction by IPTG,and purified by a agarose gel FF.Human monocytic cells were stimulated by the GST-CPAF to test the production of tumor necrosis factor a(TNF-α)and interleukin-6(IL- 6)by ELISA.Inhibition of cells proliferation with GST-CPAF was assessed by MTT.The THP-1 cell apoptosis stimulated by GST-CPAF was detected by Hoechst33258 fluorescence staining,DNA fragmentation analysis and cell apeptosis was detested bv Annexin V-FITC-propidiuum iodide (PI)staining.Results The recombination protein GST-CPAF was successfully expressed with high level in E.coli,and stimulated human monocytic cells to produce proinflammatory cytokines including TNF-α and IL-6 in a dose-and time-dependent manner.Otherwise,the GST-CPAF inhibited the growth of human monocytic cell in a dose-dependent manner.Apoptosis with nuclear chromatin fragmentation as well as cell shrinkage was observed by fluorescent staining and microscopy,DNA ladders in apoptosis cells were detected after 24 h with the GST-CPAF.Conclusion The GST-CPAF from Chlamydophila pneumoniae can induce the secretion of proinflammatory cytokines TNF-α and IL-6 by human monocytic cells,and inhibited the proliferation of THP-1 cell and apoptosis in vitro.

We investigated the effects of IFN-γ on Chlamydia psittaci (Cps) infection.HeLa cells were treated with different concentrations of recombinant human IFN-γ (5 ng/mL,25 ng/mL,50 ng/mL) after infecting with C.psittaci 6BC,then the number and morphology of C.psittaci inclusion bodies were examined after 48 hours.C57BL/6J mice were intranasally infected with 2 × 106 IFUs C.psittaci 6BC,and intraperitoneally administrated with 10 μg recombinant murine interferon-γ 24 hours prior or post infection,then body weight,activity and survival rate were recorded.The histopathology of mice livers and lungs was analyzed by HE staining on day 5 or day10 post infection.And the chlamydial inclusion bodies were titrated in the lung homogenates of mice sacrificed on day 5 after infection.The inclusion body numbers of recombinant human IFN-γ treated groups (by 5ng/mL,25ng/mL,50ng/mL) were significantly less than that in the control group (23.8±5.1)× 106,(10± 3.58) × 106,(8.0±2.22) × 106,(43.3±11.05)× 106,respectively).And the morphology of inclusion bodies in IFN-γ treated HeLa cells was irregular and much smaller.We also found that IFN-γ could significantly improve the survival rate,reduce acute clinical manifestations and pathological injurery of lung and liver in C.psittaci respiratory tract infected mice model.So we summarized that IFN-γ can mediate strong immunological protection during acute C.psittaci early infection.

Objective To analyze the clinical efficacy of Yanshu compound sophora flavescens injection (Yanshu injection) combined with paclitaxel and cisplatin (TP) regimen in the treatment of advanced non-small cell lung cancer (NSCLC). Methods One hundred and sixty-two patients with NSCLC admitted to the Department of Oncology of Danyang Hospital of Traditional Chinese Medicine from May 2014 to June 2018 were enrolled, all of their definite diagnosis was based on pathological or cytological examinations, and they were divided into two groups by the administration type, 81 cases in each group. The western medicine treatment group (western group) was treated with TP regimen alone, 3 weeks constituting 1 therapeutic course, and 3 consecutive courses were treated; based on the treatment of western drug TP regimen, the integrated traditional Chinese and western medicine treatment group (combined group) was additionally and simultaneously given Yanshu injection for consecutive 2 weeks. After treatment, the clinical efficacy and incidence of adverse reactions of the two groups were observed. Results The total effective rate in the combined group was significantly higher than that in western group [60.49% (49/81) vs. 40.74% (33/81), P <0.05]; the incidences of adverse events: alopecia, thrombocytopenia, neurotoxicity, leukopenia, nausea and vomiting, and gastrointestinal reaction in the combined group were obviously lower than those in western group [alopecia: 24.69% (20/81) vs. 60.49% (49/81), thrombocytopenia: 23.45% (19/81) vs. 40.74% (33/81), neurotoxicity: 14.81% (12/81) vs. 34.57% (28/81), leukopenia: 17.28% (14/81) vs. 62.96% (51/81), nausea and vomiting: 14.81% (12/81) vs. 60.49% (49/81), gastrointestinal reaction: 27.16% (22/81) vs. 62.96% (51/81), all P < 0.05]. Conclusion Yanshu injection combined with TP regimen is obviously effective in the treatment of advanced NSCLC, thus it is worthy to be applied clinically.

ObjectiveTo explore the characteristics of eye collaterals of patients with hepatic Wilson's disease （WD） and intervention effect of Gandouling （GDL） based on digital diagnosis information on eyes， in order to provide references for the clinical efficacy of GDL in the treatment of patients with hepatic WD. MethodEighty-two patients with hepatic WD （WD group） and 68 healthy volunteers （normal control group， NC） were enrolled. Eye images of all subjects were collected by the ophthalmoscope， followed by digital extraction of eye collateral characteristics. Differences in main eye collateral characteristics between the WD group and the NC group were analyzed. Pearson correlation test was used to analyze the eye collateral characteristics， serological indexes of liver damage ［alanine aminotransferase （ALT），aspartate transaminase （AST），and albumin （ALB）］， and Child-Pugh score in patients with hepatic WD. The 82 patients with hepatic WD were divided into a treatment group （42 cases） and a control group （40 cases）. All patients were treated with sodium 2，3-dimercapto-1-propane sulfonate （DMPS）， while those in the treatment group received additional GDL Tablets. One course of treatment contained eight days， and the treatment lasted six courses. The effects of GDL intervention on the levels of ALT，AST， and ALB，as well as the eye collateral characteristics score and Child-Pugh score were evaluated. Result① Compared with the NC group，the WD group had higher scores of "Qiu" （P<0.05） and "Xuemai" （P<0.05），which are the characteristics of eye collaterals. ② The levels of ALT and AST， and Child-Pugh score in patients with hepatic WD were positively correlated with the scores of "Qiu" and "vessel"，while the ALB level was negatively correlated with the "bulge" and "Xuemai" scores. ③ Compared with the conditions before treatment，the serum ALT and AST levels，Child-Pugh score，and the "Qiu" and " Xuemai " scores of the two groups were reduced （P<0.01），and the ALB levels were increased （P<0.01）. ④ Compared with the control group after treatment，the treatment group showed improved levels of ALT and ALB and the scores of "Qiu" and "Xuemai" （P<0.05）and better Child-Pugh score （P<0.01）. There was no significant difference in AST between the two groups. ConclusionCompared with healthy people，hepatic WD patients showed higher scores of "Qiu" and "Xuemai"， indicating that patients with hepatic WD were mainly characterized by dampness and heat，and the scores of "Qiu" and "Xuemai" were positively correlated with the severity of liver damage. Meanwhile，copper-expelling therapy combined with GDL can significantly improve the liver function of WD patients and improve the clinical efficacy.