Objective To investigate impacts of chronic hepatitis B virus (HBV) co-infection on immune responses and clinical manifestations in acute infection of dengue virus type 1.Methods The serum levels of interferon (IFN)a,IFNβ,IFNγ,interleukin (IL)-10 and tumor necrosis factor (TNF)α of 310 dengue serotype 1 (DENV1)-infected patients were assessed by enzyme linked immunosorbent assay (ELISA),among which 8％ (25/310) were chronic HBV co-infection.Meanwhile,serum samples from 41 healthy adults and 47 chronic HBV infected subjects were recruited as controls.Comparisons among groups were done by one factor analysis of variance and correlation analysis of results was done by spearman rank correlation test.Results The serum level of IFN-α [(95.1 ± 279.3) pg/mL] was significantly higher than IFN-β[(2.8 ± 16.2) pg/mL] during acute dengue infection,while IFN-α level [(86.5±358.1) pg/mL] reduced in patients with HBV co-infection.The secretion kinetics of IFNα,IFNγ (pro-inflammatory cytokine) and IL-10 (anti-inflammatory cytokine)were analyzed.The medians of IFNα,IFNβ and IL-10 level were elevated to peak on day 2,day 3-4and day 6 after fever onset,respectively.Additionally,IFNα levels in patients with only dengue infection were negatively correlated with the platelet counts and serum alanine aminotransferase levels (r=-0.2327,0.2122,both P＜0.01).Conclusions Chronic HBV co-infection alters human immune responses elicited by acute dengue viral infection.Moreover,IFNα secretion may be associated with hemorrhagic tendency,while protective against inflammatory damage of liver.

Objective To investigate the effect of emphysema and intermittent hypoxia (IH) on the hepatic oxidative stress and inflammatory injury in rats. Methods Sixty male Wistar rats were randomly assigned to 4 groups, control group (A), emphysema group (B), IH group (C) and emphysema+IH group (D). Group A was normally fed. Group B was exposed to smoke, 30 min per time, twice everyday. Group C was exposed to 5%O2 30 s/Air 90 s for 8 h/d. Group D was exposed to smoke twice, about 30 min each time, and exposed 5%O2 30 s/Air 90 s for 8 h/d. After continues exposure for 8 weeks, five rats in each group were randomly selected for arterial blood gas analysis. The tissue blocks of liver was obtained for pathologi-cal scoring and measurements of liver oxidative stress in the rest 10 rats of each group. HE staining was used to calculate the mean lining interval (MLI) and mean alveolar number (MAN). The hepatic inflammatory factor interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), superoxide dismutase (SOD) activity, catalase (CAT) activity and malondialdehyde (MDA) con-centration were measured in four groups. Results Characteristics of emphysema were found in group B and group D. The values of MLI were significantly higher in Group B and group D than those of group A and group C (P<0.05). The values of MAN were significantly lower in group B and group D than those of group A and group C (P<0.05). The levels of SOD and CAT were significantly lower in group B, group C and group D than those of group A (P<0.05). And the levels of SOD and CAT were significantly lower in group D than those of group B and group C (P<0.05). The values of liver MDA were signifi-cantly higher in group B, group C and group D than those of group A, and the values were significantly higher in group D than those of group B and group C (P<0.05). The liver histological scores and the levels of IL-6 and TNF-αwere signifi-cantly higher in group B, group C and group D than those of group A, and the values were significantly higher in group D than those of group B and group C (P<0.05). Conclusion Emphysema and IH have synergistic action in causing hepatic oxidative stress and inflammation.

Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

Objective To measure the mRNA expression of immediate-early gene c-fos in endothelial cells by the use of different intermittent hypoxia (IH) protocols. Methods A gas control delivery system was homemade, which pro-duced IH/re-oxygenation(ROX) environmental exposure. The endothelial cells were exposed to IH/ROX cycles, and were di-vided into three groups (A, B and C). There were five sub-groups in each group. Group A included intermittent normoxia group, standard incubator control group, continuous hypoxia group (CH), 1.5%O2 IH group and 10%O2 IH group. There were different frequency IH of 1.5%O2 and 10%O2 sub-groups in group B and group C. Hypoxia time was 15 s. The intermittent hypoxia cycle was 60 and the ROX time was different. The total cycle time was different, including 1.5 h group, 3 h group, 5 h group, 6.5 h group and 9.5 h group. The mRNA expression of immediate-early gene c-fos was measured by real-time PCR. Results There was no significant difference in the level of c-fos mRNA between intermittent normoxia group and CH group. The expression of c-fos mRNA was significantly higher in 1.5%O2 IH group and 10%O2 IH group than that of intermit-tent normoxia group, and there was a higher expression level of c-fos mRNA in 1.5%O2 IH group than that of 10%O2 IH group (P＜0.01). It was found that the mRNA expression of c-fos increased gradually at first, and then gradually decreased. The expression of c-fos mRNA was significantly higher in 5 h group than that of other groups (P＜0.01). Conclusion The mRNA expression of immediate-early gene c-fos is increased after exposing to IH/ROX in endothelial cells, which is closely related with IH extents and frequencies.

Objective To investigate the effect of adenovirus-mediated IL-24 (Ad-IL-24)gene expression on the apoptosis in a human squamous cell carcinoma cell line COLO 16, and to explore the underlying molecular mechanism. Methods Cultured COLO 16 cells were divided into two groups to be transfected with an adenovirus vector carrying the IL-24 gene (Ad-IL-24 group)or green fluorescent protein (Ad-GFP group), while those receiving no treatment served as the control group. After culture for different durations, qPCR was performed to quantify IL-24 gene expression, methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of COLO 16 cells, flow cytometry to detect the apoptosis of COLO 16 cells, laser scanning confocal microscopy (LSCM) to observe the morphological changes of COLO 16 cells, Western blot to determine the levels of Bax and Bcl-2 proteins and to evaluate the activation of caspase-3, qPCR to determine the levels of Bax and Bcl-2 mRNAs, an immunofluorescence assay to observe the expression of Bax and Bcl-2 proteins. Statistical analysis was carried out by a two-sample t-test with the SPSS 19.0 software. Results MTT assay showed that the proliferation of COLO 16 cells in the Ad-IL-24 group was significantly inhibited as early as 4 days after the transfection; thereafter, the inhibitory effect increased in a time-dependent manner, and peaked on day 6(P<0.05). However, there was no significant difference in cellular proliferative activity between the Ad-GFP group and control group (P>0.05). Flow cytometry revealed that the apoptosis rate was significantly higher in the Ad-IL-24 group(13.10%± 0.92%)than in the control group(3.69%± 0.36%, P<0.05)and Ad-GFP group(3.39%± 1.06%, P<0.05), but no significant difference was observed between the control group and Ad-GFP group (P>0.05). LSCM demonstrated that the apoptosis of COLO 16 cells was accelerated in the Ad-IL-24 group. The immunofluorescence assay, Western blot and qPCR all showed that the mRNA and protein expressions of Bax were increased, but those of Bcl-2 were decreased in the Ad-IL-24 group compared with the Ad-GFP group and control group. Moreover, Western blot showed a protein band that could specifically bind to the anti-cleaved caspase-3 antibody in the Ad-IL-24 group, but not in the Ad-GFP group or control group. Conclusions Ad-IL-24 can induce apoptosis in human COLO 16 squamous cell carcinoma cells, probably by up-regulating Bax expression, down-regulating Bcl-2 expression, and activating caspase 3.

Objective To explore the effect of intermittent hypoxia (IH) on RhoA/ROCK pathway in lung and on the muscularization in pulmonary vascular in rat model. Methods Wistar rats (n=40) were randomly divided into two groups:the normal oxygen control group (n=20) and the IH group ( n=20). For 4 weeks, rats in control group and IH group were ex?posed to intermittent normal oxygen (21%O2) or IH (5%-21%O2) respectively. Then, mRNA transcription and protein trans?lation levels of RhoA/ROCK were examined by Real-time PCR and Western blot. Expression of proliferation cell nuclear an?tigen (PCNA) andα-smooth muscle actin (SM-α-actin ) of lung and pulmonary artery were detected by immunohistochemis?try. Results RhoA mRNA transcription level(0.463 ± 0.067 vs 0.182 ± 0.040), ROCK mRNA transcription level(0.384 ± 0.062 vs 0.192 ± 0.052), RhoA protein expression level(0.827 ± 0.065 vs 0.424 ± 0.075)and ROCK protein expression level (0.488±0.088 vs 0.336±0.102)were higher in IH group than those in control group(P<0.05);Levels of PCNA in lung tissue [(54.67±1.80)%vs (9.14±0.91)%], PCNA in pulmonary artery [(49.40±1.21)%vs (8.38±1.13)%], SM-α-actin in lung tis?sue [(42.66±1.63)%vs (35.44±1.41)%] and SM-α-actin in pulmonary artery [(62.62±2.53)%vs (45.54±2.58)%] were also higher in IH group than those in control group(P<0.05). Conclusion Rho/ROCK pathway may play an important role in developing pulmonary hypertension (PH) associated with IH;and IH can promote the muscularization in pulmonary vascular to accelerate PH.

Rosmarinic acid (RA), a phenolic acid, is one of the important secondary metabolites produced in Salvia miltiorrhiza. To observe the influence of salicylic acid (SA), an elicitor, on the synthesis of RA and related enzymes, we treated the cell suspension cultures of S. miltiorrhiza with SA and L-a-aminooxy-beta-phenylpropionic acid (AOPP), a competitive inhibitor of tyrosine aminotransferase (TAT). Under this condition, the activities of related enzymes, such as phenylalanine ammonia-lyase and TAT were traced and assayed; the accumulative amount of RA was measured. The results showed that the PAL activity reached the peak at 4 h, 124% higher than that of the control, and the content of RA reached its maximum ((5.914 +/- 0.296) mg/g dry weight) at 8 h, after treated by 6.25 mg/L SA on day 6 of the suspension culture. The results of treatment with 0.1 micromol/L AOPP showed that AOPP affected little on the TAT activity, while the PAL activity was significantly influenced, with 44% lower than that of the control at 6 h. Meanwhile, the reduced accumulation of RA ((4.709 +/- 0.204) mg/g dry weight) paralleled with the decrease in PAL activity. The co-treatment by 0.1 micromol/L AOPP and 6.25 mg/L SA relieved the restriction imposed by AOPP on PAL, and made the cell cultures accumulate more RA than sole treatment with AOPP, indicated that SA induced the accumulation of RA in suspension cell culture of S. miltiorrhiza, and the rate-limiting effect of PAL was stronger than TAT.
Cell Culture Techniques ; methods ; Cinnamates ; metabolism ; Depsides ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Cells ; metabolism ; Salicylic Acid ; pharmacology ; Salvia miltiorrhiza ; cytology ; growth & development ; metabolism ; Suspensions ; Tyrosine Transaminase ; metabolism

Objective:To summarize the genetic and phenotypic features of MORC family CW-type zinc finger 2 (MORC2) gene-related neuropathy in Chinese patients. Methods:The clinical and whole exome sequencing data of MORC2 gene-related neuropathy families with a definitive genetic diagnosis were collected from the Third Xiangya Hospital of Central South University between 2010 and 2023. Literature involving Chinese families with MORC2 gene-related neuropathy was extensively reviewed to provide a comprehensive summary of the genetic and phenotypic spectrum of the disease. Results:A total of 10 families with MORC2 gene-related neuropathy were identified and analyzed. Six different heterozygous pathogenic variants in the MORC2 gene were observed among these families, including the novel variant c.1330G>C (p.G444R) that had not been previously reported. Six families presented as axonal Charcot-Marie-Tooth disease caused by variants in the MORC2 gene (CMT2Z) phenotype with childhood or adult onset, and carried variants c.754C>T (p.R252W), c.1199A>G (p.Q400R), c.1330G>C (p.G444R), or c.1396G>A (p. D466N); 3 families manifested as severe spinal muscular atrophy (SMA)-like phenotype with infantile onset, all carried c.260C>T (p.S87L); 1 family carried c.1181A>G (p.Y394C), presented as DIGFAN syndrome phenotype with infantile onset combined with mental and motor retardation. Systematic review showed 8 Chinese families carried pathogenic variants of the MORC2 gene, among which 5 families were associated with the CMT2Z phenotype, carrying c.754C>T (p.R252W), c.1079A>G (p.E360G), c.1220G>A (p.C407Y), or c.1397A>G (p.D466G); 1 family was associated with SMA-like phenotype, carrying c.260C>T (p.S87L); and 2 families were associated with DIGFAN syndrome, carrying c.79G>A (p.E27K) and c.292G>A (p.G98R). Conclusions:A novel pathogenic variant c.1330G>C (p.G444R) of the MORC2 gene associated with the CMT2 phenotype is reported. Eleven pathogenic variants of the MORC2 gene have been reported in the Chinese patients to date, and c.754C>T(p.R252W) may be the most common. Patients with MORC2 gene-related neuropathy carrying different variants present with significant clinical heterogeneity, manifesting as CMT2Z, early-onset severe SMA-like myasthenia, or DIGFAN syndrome.

Objective To find out the epidemic characteristics of brucellosis in Harbin and provide scientific evidences for development of countermeasures.Methods The brucellosis surveillance data were collected from the "National Diseases Surveillance Information Management System" and the municipal surveillance results from 2013 to 2017 in Harbin;the place,time and population distribution characteristics of brucellosis were analyzed by descriptive epidemiological methods.Results A total of 2 909 cases of brucellosis were reported in Harbin from 2013 to 2017,and the incidence of brucellosis declined from 5.98 per 100 000 to 4.53 per 100 000,the epidemic situation was mainly sporadic,and an outbreak caused by a live Brucella vaccine (A19 strain) was reported in 2017;all eighteen districts (counties or cities) in Harbin had reported epidemic in the past five years,Shuangcheng,Yilan,Binxian and Bayan had a large number of cases,accounted for 53.76％ (1 564/2 909) of the total number of patients in Harbin;the time of onset focused on March to July,which accounted for 57.17％ (1 663/2 909) of annual incidence;age distribution,mainly young adults aged 30 to 59,accounted for 72.57％ (2 111/2 909);gender distribution predominandy male,accounted for 75.42％ (2 194/2 909);and occupational distribution predominantly farmers,accounted for 86.28％ (2 510/2 909).In the municipal surveillance stations from 2013 to 2017,7 005 cases were surveyed,of which 2 141 cases received serological test,the positive rate was 3.32％ (71/2 141),except 2016,the serum positive rate was decreasing year by year (x2 =13.927,P ＜ 0.05),and the monitoring results of municipal monitoring stations can better reflect the epidemic trend.Conclusions The incidence of brucellosis in Harbin is decreasing year by year.However,the outbreak caused by the new vaccines should be given high attention and we should strengthen prevention and control of the disease.

Stress urinary incontinence (SUI) following prostatectomy is an important and common problem in male, which can significantly reduce the quality of life. Urethral sling is a promising treatment for male SUI. It is easy to operate and can avoid the insertion of prosthesis, making it highly acceptable to patients. In recent years, with the advancement of technology, several kinds of slings have been put into clinical application and have achieved great outcomes. This article reviews the application of urethral sling in the treatment of SUI after prostatectomy.