Objective To investigate the association between inflammatory biomarkers and survival in patients with stable chronic obstructive pulmonary disease(COPD).Methods A total of 1038 patients with stable COPD from January 2008 to December 2009 were included in a prospective cohort study.Clinical characteristics,pulmonary function tests,6 min walk test and a modified British Medical Research Council dyspnea scale (MMRC) were completed.Fasting blood was obtained to detect inflammatory biomarkers including neutrophils,C-reactive protein(CRP),fibrinogen,TNF-a,IL-6 and IL-8.Participants were evaluated every 3 months,all-cause mortality was used as the end event.Results 120 patients (9.2％) died in the period of follow-up.Kaplan-Meier analysis showed that 1 year,2-year-and 3-year survival rates were 94.4％,88.3％ and 84.0％,respectively.Compared with survivors,those who died had a higher level of inflammatory biomarkers.Cox multivariate regression analysis demonstrated that the independent risk factors for death were neutrophils (HR:1.262,95％CI:1.143-1.512,P=0.035),CRP (HR:1.234,95％CI:1.097-1.624,P=0.029),fibrinogen (HR:1.327,95％CI:1.141-1.619,P=0.026),TNF-α (HR:1.124,95％CI:1.043-1.659,P=0.045),IL-6 (HR:1,429,95％CI:1.237-1.816,P=0.014) and IL-8 (HR:1.188,95％CI:1.024-1.383,P=0.042).C statistical analysis showed that no single biomarker significantly improved the C statistic value on the base of clinical model,but it was further improved by the addition of all biomarkers (C =0.764,P =0.010).Conclusions The level of inflammatory biomarkers in the death with stable COPD is significantly increased.Age.BODE index,neutrophils,CRP,fibrinogen,TNF-α,IL-6 and IL-8 are independent risk factor for the prediction of mortality in patients with COPD.

Objective To investigate the relationship between body mass index (BMI) and long-term survival in patients with chronic obstructive pulmonary disease (COPD).Methods A retrospective cohort study was conducted in 1124 patients with completed data among 1528 hospitalized patients with COPD.Vital status was ascertained at death registry and civil affairs department.Clinical characteristics were acquired from medical record.Patients were divided into 4 groups according to the quartile of BMI:BMI＜18.9 kg/m2 (group A),BMI from 18.9 kg/m2to 22.7 kg/m2 (group B),BMI from 22.8 kg/m2 to 26.2 kg/m2 (group C),BMI＞26.2 kg/m2 (group D).The differences in survival curves were compared between groups by using log-rank test.The relationship between all-cause mortality and BMI was evaluated by Cox proportional hazards regression model.Results The median BMI of 1124 patients was 22.7 kg/m2 [(18.9-26.2) kg/m2].270 patients (24.0％) had BMI＜18.5 kg/m2.162 patients (14.4％) died during follow-up.There was significant difference in the survival curves in the four groups (x2 =7.97,P＜0.05).Survival rate was highest in group C and lowest in group A.After adjustment for other factors,BMI was an independent risk factor for predicting long-term survival.The survival rate was respectively decreased by 41 ％ and 35 ％ in group C and D as compared with group A.Conclusions Weight loss is comman in COPD patients.BMI is an independent risk factor for predicting long-term survival.BMI is easily acquired and stable,which is especially suitable to evaluate the prognosis of patients with acute exacerbation.

Objective To construct and identify the screening vector pUC18-EGFP,using EGFP as an indicator. Methods The EGFP gene was prepared by PCR and cloned into pUC18 resulting the vector pUC18-EGFP. Then DNA fragment was inserted into the MCS of pUC18-EGFP to test its practicability based on green fluorescence. Results The pUC18-EGFP was confirmed correctly by restriction enzyme analyses. The pUC18-EGFP was used to select recombinants. The green strains on the plate were confirmed by restriction enzyme and DNA analyses,which were E.coli harboring recombinants. Conclusion The screening vector PUCl8-EGFP was constructed successfully. Thus,we can select the positive clones on plates based on the green fluorescence of EGFP.

Objective To identify the Banna viruses isolated in northwestern part of Yunnan prov-ince in order to make the difference clear between the isolates and other Banna viruses isolated in other parts of Yunnan. Methods Three isolates of Banna vires isolated in 2005 and 2006 were identified by morpholo-gy, RNA-PAGE profile and molecular biologic method. Nueleotide and amino acid sequences of segment 12 of the 3 isolates were sequenced and analyzed. Results Three Banna viruses were isolated from mosquitoes collected in northwestern part of Yunnan during 2005 and 2006. Electron microscopy study showed that they are spherical with a diameter of 70 nm, no envelope but two layers of eapsid. It was found that the genome of the 3 isolates composes of 12 segments presenting band profile of 6-6 in RNA-PAGE. Nueleotide acid se-quence analysis about segment 12 showed that the identity was 99% between the 3 new isolates, 98% and 90% between the 3 isolates and the strains isolated in other parts of Yunnan, China and Indonesia, respec-tively. Phylogenetie analysis based on segment 12 gene showed that 3 new isolates clnstered in the same branch with the viruses isolated in other parts of Yunnan. The same difference of amino acids was found between Banna viruses isolated in China and Indonesia strains in the analysis of segment 12. Conclusion Banna virus strains were firstly isolated from mosquitoes collected in northwestern part of Yunnan province. Nueleotide acid sequence analysis of the 3 new isolates showed higher identity with strains isolated in other parts of Yunnan.

To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Alphavirus ; drug effects ; genetics ; metabolism ; Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; High-Throughput Screening Assays ; methods ; Luciferases ; genetics ; metabolism

Objective :To construct hepatocarcinoma specific IL-1? anti-sense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods:Murine IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 under the regulation of minimal alpha-feto protein (AFP) promoter and CMV enhancer were constructed,and further verified by PCR,restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2-antiIL-1? 1 or pafpIRES2-antiIL-1? 2 were divided into 3 groups:H22/mock,H22/antiIL-1?1 and H22/antiIL-1?2 group. Expression of IL-1? was detected by RT-PCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results:Hepatocarcinoma specific IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 were successfully constructed and were verified by PCR,restriction endonuclease analysis and DNA sequencing. IL-1? expression in H22 cells was down-regulated after transfected with IL-1? anti-sense RNA expression vectors,especially with the pafpIRES2-antiIL-1?2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL-1?2 mice was obviously smaller than that in H22/mock mice,and the cytotocixity of spleen NK against H22 cells in H22/antiIL-1?1 and H22/antiIL-1?2 mice was also greatly enhanced. Conclusion:Hepatocarcinoma specific IL-1? anti-sense RNA expression vector pafpIRES2-antiIL-1? was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL-1? and increasing cytotocixity of spleen NK.

Objective To evaluate retrograde transpopliteal access for femoral-popliteal artery total occlusion with blind puncture.Methods Clinical data of 22 cases admitted from Sep 2014 to June 2016 undergoing endovascular treatment of the femoral-politeal artery occlusion with transpopliteal artery retrograde access by blind puncture were analyzed.Results A total of 22 patients underwent retrograde popliteal access with blind puncture after antegrade attempts failed.Puncture above the knee was performed in 18 cases and below the knee in 4 cases.The average increase of ABI was 0.57.Hematoma of puncture site was observed in 2 patients,other complications included pneumonia in 1 case and renal insufficiency in 2 cases.The mean follow-up time was (13 ± 5)months.Restenosis occurred in 8 patiens(36.4％)during the follow-up time.The primary patency was (86.4 ± 0.07) ％ at 6 months and (70.7-± 0.12) ％ at 12 months.There was no major amputation rate and mortality during the follow-up.Conclusions Retrograde transpopliteal access for femoral-popliteal artery occlusion with blind puncture is safe and effective.

Objective:To explore the mediating role of self-hating in the influence of adverse childhood experiences on adolescents' negative emotions.Methods:A questionnaire method was used to collect demographic variables, and 7 012 valid questionnaires were obtained from adolescents by applying the revised adverse childhood experiences questionnaire, patient health questionnaire-4, and self-hate scale from May 1 to May 30, 2022, in five high schools(90 classes) and five junior high schools(60 classes) in Rizhao city, Shandong province, China. Data entry and analysis were performed by SPSS 22.0 software.Mann-Whitney U test was used for the comparison between demographic variables and other variables, and the correlations between variables were expressed by Spearman correlation coefficient. AMOS 23.0 software was applied for testing the mediating and moderating effects of variables. Results:(1)There were significant positive correlations between adverse childhood experiences(0(2)) and negative emotion(3(10))( r=0.459, P<0.01), self-hating(2(4))( r=0.427, P<0.01). There were significant positive correlations between self-hating and negative emotion( r=0.566, P<0.01). (2) Self-hating played a mediating role between adverse childhood experience and adolescent negative emotion, and the mediating effect was 0.299, accounted for 61.27% of the total effect.(3) The mediating pathway of self-hated was moderated by gender, with girls' adverse childhood experiences( Bsimple=2.428, t=39.585, P<0.05) predicting self-hating more than boys( Bsimple=1.641, t=25.355, P<0.05). Conclusion:Adverse childhood experiences can predict adolescents' negative emotions, and self-disgusting can also affect adolescents' negative emotions.Gender plays a moderating role in the mediating pathway.

OBJECTIVETo prepare purified and concentrated coltivirus high titer antigen in order to further detect antibodies against coltivirus in serum sample of patients.

METHODSThe coltivirus in C6/36 cells was cultured and harvested at different time, and the titer was titrated. The virus was purified and concentrated by polyethylene glycol (PEG), and stored at -20 degrees and 4 degrees, with and without glycerol, respectively, then the titer of coltivirus antigen was tested by indirect ELISA. By using the antigen, coltivirus antibodies in serum samples from both suspected Japanese encephalitis (JE) and viral encephalitis (VE) patients were detected.

RESULTSThe highest titer of coltivirus was found at 3-4 weeks of culturing. The antigen titer could be maintained at least for 6 months, especially antigen with glycerol either at 4 degrees or at -30 degrees even for two years. Totally 1141 serum samples from patients diagnosed clinically as JE and VE were tested. The results showed that 130 samples were coltivirus IgM antibody positive and the average positive rate was 11.4% (130/1141). Among 41 samples of paired-serum from patients in Guangzhou Children's Hospital, 9 samples were positive, the positive rate was 22.0% (9/41) in which 5 samples were diagnosed clinically as VE.

CONCLUSIONSStable and purified coltivirus antigen was obtained in order to test coltivirus antibodies as well as development of kits. Coltivirus probably can cause summer-autumn encephalitis in China.

Antibodies, Viral ; blood ; Antigens, Viral ; isolation & purification ; Cell Line ; Coltivirus ; immunology ; Cryopreservation ; methods ; Enzyme-Linked Immunosorbent Assay ; Humans ; Reoviridae Infections ; blood

The treatment of bi （痹） disease in The Inner Canon of Yellow Emperor （《黄帝内经》） is mainly based on acupuncture therapy. There are differences in needling depth， angle， needling techniques， number of needles and the selection of needle instruments. By reviewing literature， it is found that in The Inner Canon of Yellow Emperor， five body constituents （skin， vessel， flesh， sinew， bone） disease location differentiation is taken as the principle for the diagnosis and treatment of bi disease， guiding the needle to the lesion level where the disease is located， and according to the severity of the disease， the characteristics of pathological qi and other factors， the choice of specific acupuncture method and needles are made. This paper summarized and grouped the 17 kinds of acupuncture methods for the treatment of bi disease according to the different five body consitutuents disease location. For needle instruments， filiform needles which can softly unblock and regulate qi are often used in the treatment of bi disease. Lance needles are good at treating vessel bi with the function of clearing blood and moving qi. Round-sharp needles and fire needles are applicable for sinew bi， among which fire needles are especially good at that induced by cold. Long needles have advantages for deep-seated bi disease due to their long needle body. The puncturing method should be in accordance with the needle instruments. The five body instituents disease location differentiation and treatment is a unique system of acupuncture in the treatment of bi disease， which is worthy of more inheritance and application.