OBJECTIVE:To prepare lisinopril-loaded polyvinyl alcohol(PVA)particles(LIS-PVA-P)and establish its quality control method. METHODS:LIS-PVA-P was prepared by spray-drying process with PVA as carrier. The preparation was detected in terms of morphology,particle size,span,drug-loading capacity,encapsulation and in vitro dissolution. RESULTS:The preparation assumed sphere with porous surface. The average particle size was 17.29 ?m while drug-loading capacity 31.40%,encapsulation 94.20%,Span was 0.88,90% of the drug loads were released within 30 min. CONCLUSION:The preparation process is simple,good in repeatability and qualified in quality.

Objective:To study the relation between low density lipoprotein and macrophage myeloperoxidase.Methods:Using the reaction that MPO catalyze the oxidation of o dianisidin hydrochloride,MPO activity was determined.Using reverse transcription polymerase chain reaction(RT PCR),MPO gene expression was determined.The two methods were used to observe the relationship between low density lipoprotein and macrophage myeloperoxidase activity.Results:LDL was able to accelerate the hoist of MPO activity, but its effect is less than LPS OX LDL have no this effect When the time of LDL effect was prolonged, MPO activity was enhancing little by little When the concentration of LDL effect was increased, MPO activity was enhancing to max little by little, then it come to plateau Conclusion:LDL non especially induced the hoist of MPO activity and the swelling of MPO secretion The reason of the hoist of MPO activity was likely to make MPO into action and enhance its secretion, but not MPO gene expression

Reform was made on traditional education mode based on the criterion of undergraduate medical education at home and aboard.The reform includes the changes in teaching content,teaching methods and assessment methods in an aim to establish independent learning mode,cultivate students' self-study ability,initiative spirit and innovation ability.

ObjectiveTo investigate the effects of different laboratory test reagents for hepatitis C virus (HCV) antibody through a comparative analysis. MethodsA total of 207 samples which tested positive by four anti-HCV screening reagents commonly used in the laboratories in China (Kehua, Xinchuang, Wantai, and Abbott) were included. HCV RNA nucleic acid amplification (NAT) was performed, and if NAT results were negative, recombinant immunoblot assay (RIBA) was performed for further confirmation. The test results of these four screening reagents were compared, and their S/CO values and true positive rates were analyzed. ResultsOf all the 205 samples testing positive by any one reagent, 191 (93.2%) tested positive by the four reagents, and 14 (6.8%) were tested inconsistently by the four reagents. The positive predictive values of Xinchuang, Kehua, Wantai, and Abbott reagents were 88.2% (180/204), 93.8% (180/192), 91.4% (180/197), and 90.0% (180/200), respectively. The S/CO thresholds with a positive predictive value of ≥95% for Xinchuang, Kehua, Wantai, and Abbott reagents were 9.0, 4.0, 5.0, and 7.0, respectively. ConclusionXinchuang, Kehua, Wantai, and Abbott reagents have significantly different S/CO thresholds with a positive predictive value of ≥95%, which are significantly different from those in other domestic laboratories. Each laboratory should establish an applicable S/CO threshold with a positive predictive value of ≥95%, in order to reduce the sample size for confirmatory test.

During the Corona Virus Disease 2019 pandemic,the World Health Organization issued guidelines suggesting that antibiotics should not be used in patients with suspected or confirmed mild novel coronavirus infection unless there is clinical suspicion of bacterial infection.However,recent statistical studies have pointed out that the proportion of non-essential antibiotics used by patients infected with the novel coronavirus is very high,resulting in a large number of bacteria resistances,causing tens of thousands of lives and huge economic losses.The role of antibiotics in the treatment of diseases caused by drug-resistant bacteria is increasingly limited,and it has become an urgent social issue in the post-epidemic era.In recent years,with the progress of biotechnology and the wide application of monoclonal antibodies in the field of tumor research,the development of monoclonal antibodies for the treatment of multidrug-resistant bacterial infections has gradually attracted attention.At present,certain progress has been made in laboratory research,and the clinical trials of some products have also achieved good results,but they have not been widely used in clinical practice.This article reviews the types,advantages and related clinical trials of antimicrobial monoclonal antibodies,and looks forward to the future development and challenges.

OBJECTIVETo explore low density lipoprotein (LDL) oxidation by macrophage myeloperoxidase (MPO) at molecular level.

METHODSUsing a mouse macrophage model, we examined the relationship between LDL oxidation and macrophage MPO by measuring macrophage MPO activity, LDL oxidation products, MPO gene expression and cellular orientation of LDL oxidation.

RESULTSMPO gene expression increased to its maximum gradually when the concentration of LDL was increased, and then maintained at that level. NaN(3) inhibied the elevation of MPO activity and LDL oxidation, which was LDL concentration-dependent. After the composition of macrophage membrane was roughly analyzed, it was determined that the contents of MPO and LDL in 5% sucrose were 7.667 and 21 times higher than those in 10% sucrose, respectively.

CONCLUSIONLDL is attached to the "microdomain" of the macrophage membrane in which LDL is oxidized by MPO.

Animals ; Lipoproteins, LDL ; metabolism ; Macrophages ; metabolism ; Mice ; Oxidation-Reduction ; Peroxidase ; genetics ; metabolism

Objective:To investigate the effect of ubiquitin binding enzyme 2T (UBE2T) on the radiosensitivity of lung adenocarcinoma and unravel its possible mechanism.Methods:A total of 45 patients pathologically diagnosed with different stages of lung adenocarcinoma and treated with radiotherapy in the Second Affiliated Hospital of Zunyi Medical University from March, 2019 to December, 2021 were enrolled, and the efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST1.1). All patients were divided into radiosensitive group ( n=25) and radioresistant group ( n=20). Radiosensitive group was complete remission (CR)+partial remission (PR), and radioresistant group was stable disease (SD) + progression disease (PD). Immunohistochemistry (IHC) was used to calculate the score based on the staining intensity and the number of positive cells. Chi-square test was combined to analyze the correlation between the expression level of UBE2T in paraffin specimens of lung adenocarcinoma patients and the radiosensitivity of patients. Lentivirus UBE2T-interfered (UBE2Tsh) A549 and UBE2T-overexpressed SPC-A-1 lung adenocarcinoma cells and their respective controls were constructed for irradiation and colony formation assay. The survivor fraction curve was fitted by single-hit multi-target model. The DNA double-strand break (DSB) marker γH2AX foci were detected by immunofluorescence (IF). The expression levels of UBE2T, γH 2AX and Rad51 proteins were detected by Western blot. Cell cycle and apoptosis rate of A549 were determined by flow cytometry. Binary variables were statistically analyzed by Fisher's exact probability method and measurement data were assessed by t-test. Results:High-expression level of UBE2T was correlated with the radiosensitivity of lung adenocarcinoma patients ( P<0.05). UBE2Tsh improved the radiosensitivity of A549 lung adenocarcinoma cells, and the sensitizing enhancement ratio (SER) was 1.795. UBE2T overexpression decreased the radiosensitivity of SPC-A-1 lung adenocarcinoma cells with an SER of 0.293. γH2AX foci number per cell were significantly increased in UBE2Tsh A549 cells after irradiation ( P<0.01) . Compared with the control group, the expression level of γH2AX protein was up-regulated ( P<0.01)and that of Rad51 protein was down-regulated in UBE2Tsh A549 cells after radiation ( P<0.001). Compared with the control group, the expression level of γH2AX protein was down-regulated ( P<0.05) and that of Rad51 protein was up-regulated in UBE2T overexpressed SPC-A-1 cells ( P<0.001). The proportion of UBE2Tsh A549 cells in G 2 phase was decreased ( P<0.01) and cell apoptosis was increased ( P<0.001). Conclusions:UBE2T might promote the radioresistance of lung adenocarcinoma cells by enhancing DNA DSB repair induced by radiotherapy, inducing cell cycle G 2 phase arrest, and reducing cell apoptosis.

Aim To investigate the effect of theaflavin on oxidized low density lipoprotein(ox-LDL)-induced foam cell formation and oxidative stress in THP-1 macrophages and its mechanism.Methods THP-1 derived macro-phages were pretreated with 50 μmol/L theaflavin and(or)10 μmol/L nuclear factor erythroid 2-related factor 2(NRF2)inhibitor ML385,then 100 mg/L ox-LDL was added to the cells for 24 h to establish the foam cell model.The effect of theaflavin on THP-1 macrophages viability was evaluated by CCK-8 assay and LDH release.The expression of inflamma-tory cytokines interleukin-6(IL-6),interleukin-1 β(IL-1β),tumor necrosis factor-α(TNF-α)were analyzed by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot.The release of inflammatory cytokines were detected by enzyme linked immunosorbent assay(ELISA).Intracellular lipid accumulation was detected by Oil red O staining,and lipid absorption was observed by DiL-labeled oxidized low density lipoprotein(DiL-ox-LDL)staining.Re-active oxygen species(ROS)level was detected by DCFH-DA probe.The expression of lipid uptake,cholesterol efflux and oxidative stress-related proteins were detected by Western blot and RT-qPCR.Results Treatment with 100 mg/L ox-LDL significantly decreased cell viability and cholesterol efflux-related protein expressions,increased lipid uptake,ac-cumulation and lipid uptake-related protein expressions,and significantly promoted inflammation and ROS level,as well as the expressions of myeloperoxidase(MPO),NADPH oxidase 2(NOX2)in THP-1 macrophages(all P＜0.05).After pretreatment with theaflavin,cell viability was increased,intracellular lipid uptake,accumulation and lipid uptake-related protein expressions were significantly reduced,cholesterol efflux-related protein expressions were significantly increased,the expression and release of IL-6,IL-1β and TNF-α were significantly decreased,ROS level was significantly decreased,and the expression of MPO and NOX2 were decreased(all P＜0.05).Pretreatment with theaflavin effectively alleviated intracellular oxidative stress by altering the expression of NRF2,heme oxygenase-1(HO-1)and Kelch-like ECH-associated protein 1(KEAP1)in NRF2 signaling pathway,and enhanced the translocation of NRF2 into the nucleus.After pretreat-ment with ML385,the expression levels of NRF2,HO-1,KEAP1 and CD36 were significantly decreased.Conclusion Theaflavin can significantly inhibit ox-LDL-induced foam cell formation,inflammation,and oxidative stress through NRF2/HO-1 signaling pathway in THP-1 macrophages.

Objective: To investigate the differences in enolase and laminin levels in vaginal epithelial tissues between mice successfully infected withGardnerellaand mice not infected with Gardnerella, providing information for further exploration of the correlation between enolase and laminin levels and the incidence of bacterial vaginosis. Methods: Gardnerella strains isolated, purified, and identified from vaginal secretions of patients with bacterial vaginosis were used to infect the vagina of mice and establish a mouse model of bacterial vaginosis. Successful and failed mice was defined as successful and failed groups, respectively. Differential expression of enolase and laminin in the vaginal epithelial tissue of two groups of mice was detected by Western blot. Modeling success rate was statistically analyzed, and the expression differences of enolase and laminin was compared between two groups. Results: One strain of Gardnerella vaginalis infected 10 SPF grade KM mice, 7 mice met the diagnostic criteria for bacterial vaginosis, and 3 mice failed to model, with a success rate of 70%. Western blot was used to detect protein expression levels, and the levels of laminin and enolase in the successfully modeled mouse vaginal epithelial tissue were significantly higher than those in the failed modeling group, with statistical differences between the two groups(P<0.05). Conclusion Enolase and laminin may be involved in the occurrence of bacterial vaginosis, however, further research is needed to determine the mechanisms through which they trigger the occurrence and development of the disease.

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.
Chromatography, Liquid ; Cloning, Molecular ; Escherichia coli ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; Tandem Mass Spectrometry