OBJECTIVE:To investigate the effect of STAT5 pathway inhibitor pimozide on the expressions of nitric oxide (NO)and nitric oxide synthetase(iNOS)in the model of mouse macrophage RAW264.7 inflammation induced by lipopolysaccha-ride (LPS). METHODS:RAW264.7 cells in logarithmic growth phase were divided into blank control group,drug control group (10μmol/L pimozide),model group(1μg/ml LPS)and the pimozide groups of low,middle and high doses(2.5,5 and 10μmol/L), where the corresponding cells were given pimozide 30 min before the administration of LPS,and then were cultured for 24 h. Griess method was used to determine the content of NO in the supernate of cell culture solutions of all groups,real-time quantita-tive polymerase chain reaction(RT-PCR)to determine iNOS mRNA expression,and Western blot method to determine the protein expression of iNOS and phosphorylated STAT5(p-STAT5). RESULTS:The content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells in the model group were higher than those in the blank control group,with statisti-cally difference (P<0.01). Compared to the model group,the pimozide groups of middle and high doses had lower content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells,with statistically difference(P<0.01 or P<0.05). CONCLUSIONS:STAT5 pathway inhibitor pimozide can inhibit the release of NO by inhibiting iNOS mRNA and pro-tein expressions in cells.

Objective To evaluate effects of trichostain A (TSA) on cell proliferation, cell cycles, apoptosis and invasiveness of multiple myeloma cell line U266; as well as active changes of methylation regulating proteins including DNA methyl-transferase(DNMTs), methyl-binding domain (MBD) proteins: MBD2 and MeCP2 after treated with TSA. Methods U266 cells were treated with different concentrations of TSA for 12, 24, 48 and 60 h. The proliferation activity of U266 cells was detected by MTT and the IC50 of 24 h was calculated. After U266 cells were treated with IC50, cell cycles were check out by dying with PI. mRNA of matrix metalloproteinase-2(MMP-2), bc1-2, bcl-xl and methylation regulating proteins (DNMTs, MBD2 and MeCP2) were detected by real-time PCR. FCM and Western blotting were used to measure expressions of MMP-2 and MBD2. Results MTT results revealed TSA inhibited proliferation of U266 cells in a dose-and time-dependent manner and the IC50 of 24 h was 0.07 μmol/L FCM analysis showed that TSA could arrest the cell cycle in G0/G1 and the proliferation index (PI) in U266 cells [(49.90 0.39)%]were significantly different after exposed to TSA (0.7 μmnol/L for 24h compared with that in the control cells[(55.78 0.49)%](P <0.01). After treated by TSA, the 2-△△Ct of MMP-2, bcl-2 and bcl-xl were 0.71 0.06, 5.04 0.92 and 2.95 0.35, respectively. There were great changes on mRNA of DNMT, MBD2 and MeCP2. TSA could reverse the transcription of DNMT, MBD2 and MeCP2. Conclusion TSA can arrest the U266 cell cycle in GVG, to prevent its proliferation and promote apoptosis, which maybe greatly connect with the changes of the methylation regulating proteins.

Objective: To investigate the epidemiological characteristics of pulmonary tuberculosis (PTB) among residents aged 60 years and older in Yangzhou City, Jiangsu Province from 2013 to 2022, so as to provide the evidence for the improvement of PTB prevention and control measures in this population. Methods: Data of PTB cases aged 60 years and older in Yangzhou City from 2013 to 2022 were collected from the Chinese Disease Prevention and Control Information System. The temporal, population, and regional distribution characteristics of PTB cases were analyzed using the descriptive epidemiological method. Results: A total of 8 726 PTB cases aged 60 years and older were registered in Yangzhou City from 2013 to 2022, including 4 167 cases positive for pathogenic tests, with a positive rate of 47.75%. The registered incidence rates and the positive rates of pathogenic tests of PTB among residents aged 60 years and older in Yangzhou City showed downward trends from 2013 to 2022 (both P<0.05). The average annual registered incidence rate of PTB was 83.95/105, and the average annual registered incidence rate of PTB positive for pathogenic tests was 40.09/105. The average annual registered incidence rate of PTB in males was higher than that in females (138.57/105 vs. 31.76/105, P<0.05). The registered incidence rate of PTB showed an increasing trend with age (P<0.05), with the highest rate observed in the age group of 75-<80 years (110.37/105). The top three regions with the highest average annual registered incidence rates of PTB were Jiangdu District (94.34/105), Baoying County (91.61/105) and Hanjiang District (84.93/105). Conclusions The registered incidence of PTB among residents aged 60 years and older in Yangzhou City showed a downward trend from 2013 to 2022. Particular attention should be payed to males, residents aged 75- <80 years, and the elderly in Jiangdu District and Baoying County.

To investigate the protective effects of scutellarin benzyl ester on neonatal rats' cardiomyocytes injured by ischemia and its anti-apoptosis mechanism.

Objective To establish a determination method for leuprolide acetate microspheres. Methods HPLC was performed on Inertsil ODS-SP (150×4.6 mm×5μm) with mobile phase consist of 0.1 mol/L of Ammonium Dihydrogen Phosphate(adjust its pH to 7.00±0.05 with Ammonium Hydroxide) and Acetonitrile in ratio of 3:1(V/V),and the flow rate was 1.0 mL/min.The wavelength was 220 nm and column temperature was 30℃. The injection volume was 20 μL. Results The linear range of leuprolide acetate was 20.0-160.0μg/mL (r=0.9999) with an average recovery of 99.80%,RSD=0.63%(n=9). Conclusion The method of HPLC was accurate,reliable and specific, which could be used to determinate the assay of leuprolide acetate microspheres and for quality control of microspheres.

Objective To investigate the effects of dexmedetomidine pre-treatment on pneumonocyte apoptosis and CCAAT/enhancer binding protein homologous protein (CHOP) in acute lung injury (ALI) induced by ischemia/reperfusion (I/R) during orthotopic liver transplantation in rats.Methods Forty adult male Sprague-Dawley (SD) rats were randomly divided into four groups by random number table method: sham operation group, I/R model group, dexmedetomidine low dose group and dexmedetomidine high dose group, 10 rats per group. Hepatic artery was ligated and cut off by two cuff method, and the portal vein was completely opened after donor liver transplanted into the recipient, thus, a hepatic I/R model was established. The perihepatic ligaments of rats were just separated after laparotomy in sham operation group and no other special treatment was performed. One hour prior to I/R, dexmedetomidine at a dose of 2.5μg·kg-1·h-1 and 5.0μg·kg-1·h-1, respectively, were pumped intravenously and finished within 1 hour in the rats of low dose group and high dose group. After experiment, the lung tissue was taken, and the lung wet/dry weight (W/D) ratio was determined. Pathological changes of lung tissue were observed and alveolar damage index of quantitative assessment (IQA) was tested by light microscope, and changes of ultrastructure of lung tissue were observed by transmission electron microscope. The mRNA and protein expressions of CHOP were detected respectively by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. The apoptosis in lung tissue was determined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method and apoptosis index (AI) was calculated.Results Compared to sham operation group, the lung W/D ratio (4.94±0.84 vs. 2.29±0.54), IQA [(40.52±5.15)% vs. (4.55±1.85)%] and AI [(36.57±5.85)% vs. (2.85±0.95)%] in I/R model group were significantly higher (allP < 0.01); remarkable injury of lung tissue was confirmed by light microscope and transmission electron microscope in the I/R model group. Compared to I/R model group, the W/D ratio (3.29±0.85, 2.68±0.78 vs. 4.94±0.84), IQA [(23.69±2.62)%, (15.86±3.61)% vs. (40.52±5.15)%] and AI [(25.73±3.71)%, (14.66±2.61)% vs. (36.57±5.85)%] in dexmedetomidine low and high dose groups were markedly lower (allP < 0.01); under light and transmission electron microscopes, the injury of lung tissue in these two dose groups was notably alleviated. There was a large amount of apoptotic cells of pulmonary vascular endothelium and alveolar epithelium in I/R model group, while the cell apoptosis was distinctly decreased in dexmedetomidine low and high dose groups compared to that in model group. Compared to sham operation group, the expressions of CHOP mRNA [absorbance (A) value: 0.96±0.18 vs. 0.43±0.08] and protein (gray scale: 2.79±0.74 vs. 1.02±0.27) were significantly higher in I/R model group (bothP < 0.01). Compared to I/R model group, the expressions of CHOP mRNA (A value: 0.69±0.13, 0.56±0.12 vs. 0.96±0.18) and protein (gray scale: 1.96±0.58, 1.34±0.49 vs. 2.79±0.74) were significantly lower in dexmedetomidine low and high dose groups, the decrease in dexmedetomidine high dose group being more marked (allP < 0.01).Conclusion The pretreatment of dexmedetomidine can protect lung tissue against I/R injury during liver transplantation in rats, and the mechanism may be related to the suppression of CHOP activation and alleviation of lung tissue cell apoptosis.

Objective: To evaluate the effectiveness of recombinant Mycobacterium tuberculosis fusion protein skin test (EC-ST) in screening for latent tuberculosis infection (LTBI) among HIV/AIDS patients, so as to provide insights into the applicability of EC-ST in LTBI screening among HIV/AIDS patients. Methods: From April to June 2023, HIV/AIDS patients under management and treatment in Yangzhou City, Jiangsu Province, were selected as study subjects. Basic information was collected through questionnaire surveys. LTBI was screened by EC-ST and interferon-gamma release assay (IGRA). Taking IGRA results as the diagnostic standard, the positive rate, sensitivity, specificity and consistency rate of EC-ST, and the impact of CD4+T lymphocyte (CD4) counts on the screening effect of EC-ST were analyzed. Results: A total of 523 HIV/AIDS patients were screened, including 458 males (87.57%) and 65 females (12.43%). The median age was 48.00 (interquartile range, 21.00) years. The positive rate of EC-ST was 7.27% and the positive rate of IGRA was 7.46%, with no statistically significant difference (P>0.05). The consistency rate of the two methods was 94.84%, and the Kappa value of 0.621 (95%CI: 0.489-0.752, P<0.05). The sensitivity of EC-ST was 64.10% and the specificity was 97.31%. Comparing the groups with CD4 counts <500 and ≥500 cells/μL, the consistency rates of the two methods were 95.32% and 94.44%, and the Kappa values were 0.568 and 0.650, respectively (both P<0.05). There were no statistically significant differences in the positive rates, sensitivity, and specificity of EC-ST (all P>0.05). Comparing the groups with CD4 counts <200 and ≥200 cells/μL, the consistency rates of the two methods were 96.55% and 94.62%, and the Kappa values were 0.648 and 0.619, respectively (both P<0.05). There were no statistically significant differences in the positive rates, sensitivity, and specificity of EC-ST (all P>0.05). Conclusion The effectiveness of EC-ST in screening for LTBI among HIV/AIDS patients is consistent with that of IGRA and is not affected by CD4 counts.

Objective To screen mutations in genes including ASXL1, TET2, IDH1, IDH2, SETBP1, MPL515, JAK2 exon 12 and JAK2V617 in 135 polycythemia vera (PV) patients. To assess progreasson and genomics characteristics post polycythemic myelofibrosis. Methods DNA sequencing of ASXL1(Exon12),TET2 (Exons 3-11),IDH1(Exon4),IDH2(Ex-on4),SEPBP1(Exon4),JAK2 exon 12 and MPL515 (Exon 10) genes were carried out using Sanger method. JAK2V617 muta-tion was detected by allele-specific PCR in patients with PV. In the mean time, the mutation load of JAK2V617F allele (V617F%) was evaluated by real-time PCR using Tagman MGB probe. Then, the significant of gene mutations and clinical outcomes of post-PV Myelofibrosis(PPMF)was analyzed. To study risk factors of PPMF, logistic regression were employed. Results ASXL1, TET2, IDH1, IDH2 were mutated in 7.69%(8/104), 5.26%(1/19) , 0.08%(1/120) and 0.08%(1/121) of all PV patient respectively. JAK2 was mutated in 82.22%(111/135) of PV patients with mutation rate of exon12 of 2.96%(4/135) and there were no mutation of MPL515 and SETBP1 in PV patients. ASXL1 mutation was found in 31.82%(7/22) PPMF patients. Spearman analysis showed that ASXL1 is correlated with JAK2V617F (V617F%) (rs=0.298,P=0.002). The hemo-globin was lower in patients with ASXL1 mutation than patient without mutation (wild type). Leukocyte count, V617F%>50%rate, thrombosis and PPMF were higher in patients with ASXL1 mutation than that of ASXL1 wild type(P＜0.05). ASXL1 mu-tation, V617F%>50% rate and splenomegaly were all risk factors of PPMF. Conclusion ASXL1 mutation is the risk-fac-tor of PPMF and may promote V617F%by some mechanism.

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.

Objective To predict and identify liner B-cell epitopes in the hemagglutinin ( HA) of human-infected avian-origin H7N9 influenza virus and analyze the specificity of H7 subtype.Methods Three serum samples collected at different times from the same patient who was confirmed to be infected with H7N9 influenza virus were provided by Shaoxing People’s Hospital, and one serum sample from healthy person was collected as the control.The extracellular region of HA protein was predicted by TMHMM Sever v.2.0.The potential B-cell epitopes were predicted by DNAStar Lasergene’ s Protean, BcePred and ABCpred tools, and the immunogenicity of the predicted B cell antigen epitopes was assessed by indirect enzyme-linked immunosordent assay ( ELISA ) .H7 subtype specificity was analyzed by comparing HA protein amino acid sequence with H7N9 and H1-H16 subtype influenza virus from Genbank using Clustal X 2.1 software, and Cn3D 4.3.1 software was used to detect the distribution and 3D structure of predicted epitopes on the HA protein of H7N9.Results The potential B-cell epitopes may be located in 172-183, 363-380, 452-472 and 491-506 of extracellular N-terminus of HA protein.ELISA showed that four predicted eptiopes specifically reacted with positive serums from patient.Multi-sequence alignment demonstrated that peptide 172-183 and 363-380 had higher H7 subtype specificity compared with amino acid sequences of other subtypes.Moreover, the predicted linear B-cell epitopes all located on the surface of HA protein according to the 3D structure analysis.Conclusion Four potential B-cell epitopes were identified, in which peptide 172-183 and 363-380 have higher H7 subtype specificity, and may be used in the design of epitope-based vaccines and diagnostics tests.