BACKGROUND: Preliminary study has proved that the bone marrow-derived mesenchymal stem cells (MSCs) in a rat emphysema model produced by use of trypsin alone can "homing" to the lesioned lung tissues, and participate in the formation of pulmonary arteries to promote lung tissue repair. Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) play equally a powerful role in promoting angiogenesis. OBJECTIVE: To observe the influence of bFGF, VEGF and MSCs in regeneration of pulmonary capillary and pathological repair of pulmonary emphysema rats. METHODS: Except normal control group, the remaining 5 groups of rats were exposed to tobacco smoke and received a single intratracheally instillation of porcine pancreatic elastase to induce emphysema models. Following successful modeling, rats of bFGF group were intratracheally injected with 400 U bFGF and rats of VEGF group with 2 μg VEGF, once a week for three times. MSCs group was injected 1 mL suspension of 4×10~9/L MSCs into tail vein. MSCs+VEGF group was injected MSCs into tail vein and intratracheally injected VEGF (2 ug, three times) at the same time. Model control and normal control groups were intratracheally injected with equal volume of sodium chloride. Four weeks after treatment, arterial blood gas analysis was performed to observe pathological and morphological changes of lung tissues. CD34~+ expression in lung tissues was determined using immunohistochemistry method. RESULTS AND CONCLUSION: Compared with model control group, PaO_2 values dramatically increased in VEGF group (P <0.05), while other indices remained unchanged (P > 0.05); there were no obvious changes in each index in other groups (P >0.05). Gross and microscopic observations showed that, lung was smooth, pale pink, and elastic in normal control group, with uniform size of pulmonary alveoli on cross-section; pathological changes of chronic obstructive pulmonary emphysema existed in model control group, but improved in other 4 groups. Compared with model control group, mean pulmonary alveoli number and CD34~+ relative positive area dramatically increased in bFGF, VEGF, MSCs, MSCs+VEGF groups (P < 0.05), mean linear intercept and mean alveoli area were significantly reduced (P < 0.05). No significant difference was observed in each index among these 4 groups (P > 0.05). bFGF, VEGF and MSCs could improved the pathology of pulmonary emphysema models produced by tobacco smoking and intratracheally instillation of porcine pancreatic elastase. The possible mechanism of recovering the pulmonary emphysema is the proliferation of pulmonary capillary and enlargement of pulmonary artery, improved blood flow in the lung, improved ventilation/perfusion shunt, reduced pulmonary alveolus size and volume of the lung through self-compensation.

Tissue factor pathway inhibitor (TFPI) is one of the major physiological inhibitors of the human blood coagulation cascade and may have great potential in the prevention and therapy of diseases caused by thrombus formation. In this study, recombinant human tissue factor was generated in E. coli containing a recombinant vector constructed by inserting TFPI cDNA into pGEX-2T vector. The generated recombinant TFPI (rTFPI) could be simply purified with glutathione-agarose affinity method and maintained its biological function in terms of inhibition of tissue factor and factor Xa.
Escherichia coli ; genetics ; metabolism ; Humans ; Lipoproteins ; biosynthesis ; genetics ; pharmacology ; Peptide Fragments ; biosynthesis ; genetics ; pharmacology ; Plasmids ; Recombinant Proteins ; biosynthesis ; pharmacology ; Transfection

Objective Endothelial microparticles (EMPs) are direct indicator of endothelial cell activation or apoptosis,and may also reflect endothelial inflammation,increased coagulation,and vascular tone.The aim of this study is to investigate whether EMPs would be able to evaluate systemic involvement and be a new indicator of disease activity in Beh(c)et's disease (BD).Methods Thirty-nine consecutive BD patients (who fulfilled the modified International Study Group on BD in 1990 or International Criteria for BD in 2006) and 67 age and sex-matched healthy controls were enrolled (Including 37 patients with hypertension and 30 healthy subjects).The plasma levels of EMPs were measured by flow cytometry utilizing specific labels for endothelial MPs (CD31+ and CD42b-).The measurement data of each group were expressed as-x±s,and the comparison data betwen groups were analyzed by independent sample t test and analysis of variance,Spearman/Pearson correlation analysis,P＜0.05 was statistically significant.Results The levels of circulating EMPs (CD31 + and CI42b-) were significantly elevated in the case group compared with the healthy control group and hypertension (F=6.845,P＜0.05).Moreover,BD patients plasma EMPs were positively correlated with active BD (r=0.802,P＜0.05).Vascular involvement in BD patients was higher than in patients without vascular EMPs,t=4.707,P＜0.05.Gastrointestinal involvement in BD patients was more frequent than that in patients without Gastrointestinal involvement,t=2.673,P＜0.05.Conclusion Levels of circulating EMPs are elevatedd in BD patients and correlated with disease activity in BD.Elevated EMPs may be a potential indicator to predict disease activity of BD.The plasma level of EMPs is increased,which indicats increased risk of vascular and digestive tract involvement in BD.

Objective To compare five public deep venous thrombosis(DVT)risk assessment models(RAM)in patients with acute cerebral hemorrhage,and to select one suitable assessment tool,with which medical staff are able to carry out effective individualized prevention accordingly in time.Methods In this retrospective study,256 in-patients with acute cerebral hemorrhage in the Neurology Department of the First Affliated Hospital of Wenzhou Medical University from January to December 2020 were randomly enrolled.The researchers applied five models and record the scores,including Padua RAM,Caprini RAM,Wells DVT RAM,Autar RAM and acute stroke DVT-RAM.The data were summarized and analyzed with SPSS 25.0 and R4.1.The predictive values of above RAMs are compared separately by TOPSIS(technique for order preference by similarity solution)to find out a suitable tool.Results Predictive value(Ci)ranking of five DVT-RAMs,Autar RAM,Padua RAM,Caprini RAM,acute stroke DVT-RAM and Wells DVT RAM,by TOPSIS were 0.681,0.636,0.385,0.363,0.315.Conclusion According to comprehensive comparison by TOPSIS,Autar RAM was the suitable assessment model to assess the DVT risk of patients with acute cerebral hemorrhage.It is suggested that medical staff apply the Autar risk assessment model to predict the risk of DVT in patients with acute cerebral hemorrhage.

Objective:To analyze the characteristics of HIV-1 molecular network in men who have sex with men (MSM) from 2016 to 2018 in Kunming, Yunnan province, explore the risk factors associated with HIV-1 transmission network and provide evidence for the effective implementation of intervention.Methods:A total of 540 samples of newly reported HIV-1 positive MSM were consecutively collected in Kunming from 2016 to 2018, the pol gene fragments were amplified by nested polymerase chain reaction (PCR). HIV-1 molecular networks were constructed according to the bootstrap value of the maximum likelihood evolutionary tree over 95% and the genetic distance less than 3%. The factors associated with the subjects entering network and network growth were further analyzed. Results:Among 459 successfully sequenced samples, seven genotypes were found, in which CRF07_BC (49.2%, 226/459) and CRF01_AE (40.3%, 185/459 ) were predominant. Other genotypes included URFs (4.8%, 22/459), CRF08_BC (3.1%, 14/459), CRF55_01B (1.7%, 8/459), B (0.7%, 3/459) and CRF68_01B (0.2%, 1/459). A total of 163 sequences entered the network, with an entry rate of 35.5%(163/459), forming 56 clusters with the number of individuals in the cluster was between 2 and 13. The analysis of the factors associated with entering network showed that the MSM who married and had multiple homosexual partners were more likely to be found in HIV-1 molecular networks. Multivariate logistic regression analysis showed that the number of sexual partners was the factor for the growth of HIV-1 molecular network. According to the criteria for the emergence of three or more newly reported cases in every year, six transmission clusters were judged as active transmission clusters, in which MSM who were not Kunming natives, had sexually transmitted diseases (STD), were divorced and students were the key targets of intervention.Conclusions:HIV-1 genotypes in MSM in Kunming were becoming complex, the risk factors associated with transmission networks in MSM in Kunming included being married and having multiple partners, the intervention targets in active transmission clusters included MSM who were not Kunming natives, had STD, were divorced and students. This study provided the basis for applying HIV-1 molecular networks to real-time intervention in this population.

Objective:To understand the infection status of HCV and Treponema pallidum (TP) in HIV/AIDS cases in Yunnan province,and identify the risk factors. Methods:Between January 1 and June 30 in 2020,a cross-sectional survey was conducted in Yunnan. Two enzyme-linked immunosorbent assay (ELISA) kits were used to detect anti-HCV, the positive results of both two kits indicated HCV infection. ELISA and syphilis toluidine red untreated serum test were applied to identify TP infection. Both Excel 2016 and SPSS 22.0 software were used for statistical analysis, and logistic regression model was conducted to identify the relevant factors of HCV and TP infection.Results:A total of 5 922 HIV/AIDS cases were included in this study, the infection rates of HCV and TP were 6.5% (383/5 922) and 5.8% (344/5 922) respectively. The co-infection rate of HCV and TP was 0.4% (22/5 922). The risk for HCV infection in HIV/AIDS cases was higher in younger age groups compared with age group ≥50 years (15-19:a OR=3.53;20-29:a OR=3.02;30-39:a OR=2.91;40-49:a OR=3.61), in males than in females (a OR=2.31), in the married and unmarried than in the divorced or widowed (married:a OR=1.61;unmarried:a OR=1.63), in other ethnic groups than in Han ethnic group (a OR=1.70), in people with lower education level than in people with education level of college and above (primary school degree and below:a OR=4.69;middle school:a OR=3.96), in people living in the central and western Yunnan than in people living in eastern Yunnan (central Yunnan:a OR=2.46; western Yunnan:a OR=7.08), in injection drug users than in MSM (a OR=131.08). The risk of TP infection in HIV/AIDS cases was higher in people with education level of college and primary school than in middle school degree (primary school and below:a OR=1.73;college and above:a OR=1.77), in people with other occupations than in farmers (a OR=1.39), in people living in eastern Yunnan than in people living in western Yunnan (a OR=1.75); in MSM than in people with heterosex (a OR=9.75). Conclusions:A certain proportion of HIV/AIDS cases reported between January and June in 2020 in Yunnan were co-infected with HCV and TP, many factors were associated with the co-infection. It is suggested to strengthen HCV and TP tests in HIV/AIDS cases and conduct active treatment of the co-infection.

Objective:To understand the characteristics of HIV-1 genotypes and drug resistance among men who have sex with men in Kunming in 2018.Methods:A total of 193 plasma samples were collected from the newly reported HIV-1 infected MSM in Kunming from January to December 2018. Viral RNA was extracted, and the gag, pol, env gene segments were amplified by nested PCR. HIV-1 genotypes and drug resistance were also analyzed. Subsequently, the evolutionary characteristics of CRF55_01B and CRF68_01B among MSM in Kunming were analyzed by Bayesian Markov Chain Monte Carlo method. Results:Multiple HIV-1 genotypes were identified among these 193 samples, including CRF07_BC (39.4%, 76/193), CRF01_AE (34.2%, 66/193), unique recombinant forms (URFs) (20.2%, 39/193), CRF08_BC (3.1%, 6/193), CRF55_01B (1.6%, 3/193), subtype B (1.0%, 2/193) and CRF68_01B (0.5%, 1/193). Results from the Bayesian evolutionary analysis showed that CRF55_01B started to spread locally after being imported from other provinces, while CRF68_01B was likely to have been brought in from the eastern provinces of China. Prevalence of HIV-1 drug resistant strains was 2.6%(5/190) before antiviral treatment, with mutation rates resistant to non-nucleoside reverse transcriptase inhibitors being the highest (2.1%, 4/190) among MSM in Kunming, 2018.Conclusion:The diversity of HIV-1 was increasing among MSM in Kunming. Although the resistance rate on pretreatment drug was relatively low, the emergence of multiple resistant strains to first-line antiviral drugs posed a challenge to antiretroviral therapy, in Kunming.

Objective:To assess and compare the performance of limiting-antigen avidity enzyme immunoassay (LAg-Avidity EIA) and pooling PCR in the surveillance for recent infection rates of HIV-1 in men who have sex with men (MSM).Methods:Blood samples were collected from MSM selected through snowball sampling method in sentinel surveillance in 13 prefectures of Yunnan province from 2016 to 2017. The samples were tested for HIV-1 antibody. The confirmed positive samples were tested by LAg-Avidity EIA. The negative samples were tested by pooling PCR. The recent infection rates of HIV-1 were estimated by the algorithm based on LAg-Avidity EIA and pooling PCR respectively. The two results were compared.Results:During 2016-2017, a total of 5 363 blood samples were collected from MSM, in which 407 samples were HIV-1 positive (including 177 positive tested previously) and 4 956 samples were HIV-1 negative. A total of 211 samples(91.7%) were tested by LAg-Avidity EIA, 69 were confirmed to be recent infections. A total of 4 469 samples were tested by pooling PCR, 8 were confirmed to be acute infections. The recent infection rates of HIV-1 from 2016 to 2017 estimated by LAg-Avidity EIA were 3.36% and 4.84%, and the recent infection rates estimated by pooling PCR were 3.27% and 3.02% respectively. The differences in recent infection rates of HIV-1 estimated by the two algorithms were not significant.Conclusions:The recent infection rates of HIV-1 estimated by LAg-Avidity EIA and pooling PCR in sentinel surveillance in MSM in Yunnan had good consistency from 2016 to 2017. Using the two methods might have a better stability in continuous surveillance for recent infection rates of HIV-1.