Selenium attenuates the inflammatory injury induced by tumor necrosis factor-α in AC16 cardiomyocytes / 中华地方病学杂志

Objective:To exploring the effect of selenium on tumor necrosis factor-α (TNF-α) induced inflammatory injury of AC16 cardiomyocytes.Methods:AC16 cardiomyocytes cultured in vitro were divided into control group, selenium pretreatment group (100 μg/L sodium selenite pretreatment for 4 h), 50 μg/L TNF-α group, selenium pretreatment + 50 μg/L TNF-α group, 100 μg/L TNF-α group, and selenium pretreatment + 100 μg/L TNF-α group. After 24 h of culture, the cell culture medium or cells were collected for detection. Griess method was used to detect nitric oxide (NO) level in cell culture medium. Hoechst 33342 nuclear staining was detected by fluorescence inverted microscope and karyopyknosis ratio was analyzed. The mRNA expression levels of B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-2 associated X protein (Bax) and inducible nitric oxide synthase (iNOS) were detected by real-time fluorescence quantitative PCR. The protein expression levels of nuclear factor-kappa B (NF-κB) p65 and inhibitory κB-α (IκB-α) were detected by Western blotting. Results:The levels of NO in the control group, selenium pretreatment group, 50 μg/L TNF-α group, selenium pretreatment + 50 μg/L TNF-α group, 100 μg/L TNF-α group and selenium pretreatment + 100 μg/L TNF-α group were (10.58 ± 2.32), (9.07 ± 0.73), (15.53 ± 3.97), (12.05 ± 1.11), (30.65 ± 4.16) and (19.02 ± 3.72) μmol/L, respectively. Selenium and TNF-α had main effects on NO level ( F = 37.71, 99.07, P < 0.001) and interaction effect ( F = 11.80, P < 0.001). Selenium and TNF-α had main effects on karyopyknosis ratio and Bcl-2 and Bax mRNA expression levels ( F = 56.37, 462.81, 18.04, 32.85, 18.38, 170.77, P < 0.05). Selenium and TNF-α had an interactive effect on both karyopyknosis ratio and Bax mRNA expression level ( F = 21.48, 19.96, P < 0.001). Selenium and TNF-α had main effects on iNOS mRNA expression level ( F = 129.98, 1 051.76, P < 0.001) and interaction effect ( F = 53.28, P < 0.001). Selenium and TNF-α had main effects on the protein expression levels of NF-κB p65 and IκB-α ( F = 73.65, 145.49, 710.20, 105.66, P < 0.001) and interaction effects ( F = 26.73, 197.59, P < 0.001). Conclusion:Selenium may inhibits iNOS expression through NF-κB signal system and protects cardiomyocytes from TNF-α induced inflammatory injury.

Association between single nucleotide polymorphisms of TGFβ2 gene and Keshan disease in Shaanxi Province / 中华地方病学杂志

Objective:To investigate the relationship between single nucleotide polymorphisms of transforming growth factor-β2 (TGFβ2) gene and Keshan disease (KD) in Han population of Shaanxi Province.Methods:KD region in Huangling County, Yan'an City, Shaanxi Province was selected as the investigation site in this study. Using the method of cluster random sampling, 52 families with KD in 6 administrative villages in Huangling County (Duanjiawan Village, Taoqu Village, Yaoping Village, Jianzhuang Village, Anjiao Village in Yaoping Town, and Houziping Village in Diantou Town) were selected for epidemiological investigation. According to the "Diagnosis of Keshan Disease" (WS/T 210-2011), 285 subjects were identified, including 79 patients with KD (case group) and 206 healthy controls (control group). Genomic DNA was extracted from the peripheral venous blood. The polymorphism of genetic variation of TGFβ2 gene rs6658835 was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Chi-square (χ 2) test and t-test were used to analyze the baseline data, and binary logistic regression model was used to analyze the influencing factors of KD, all samples were tested for Hardy-Weinberg equilibrium using goodness-of-fit χ 2 test, differences in genotype and allele frequencies between case and control groups were compared by χ 2 test, and logistic regression analysis was used to compare the genotype frequencies between two groups after adjusting for confounding factors. Results:Epidemiological investigation showed that there were significant differences in age and heart murmur between case group and control group ( t = 7.03, χ 2 = 9.66, P < 0.05). The analysis of binary logistic regression model showed that the influence of age on KD was statistically significant (χ 2 = 20.72, P < 0.001). The gene frequency distribution of TGFβ2 gene rs6658835 in case group and control group conformed to the Hardy-Weinberg equilibrium (χ 2 = 0.02, P = 0.900). Correlation analysis results: the difference of genotype frequency of TGFβ2 gene rs6658835 in case group (GG, GA, AA: 6.3%, 38.0%, 55.7%) and control group (GG, GA, AA: 10.7%, 43.7%, 45.6%) was not statistically significant (χ 2 = 2.78, P = 0.249). After adjustment by age, the difference of genotype frequency and dominant model of TGFβ2 gene rs6658835 in case group and control group was statistically significant (χ 2adj = 5.43, 4.86, P < 0.05), the difference of recessive model of TGFβ2 gene rs6658835 in case group and control group was not statistically significant (χ 2adj = 2.12, P = 0.145). Conclusion:TGFβ2 gene rs6658835 is associated with KD in Han population of Shaanxi Province.

An association between single nucleotide polymorphisms of IL-23R gene and Keshan disease in Northwest Chinese Han population / 中华地方病学杂志

Objective To investigate the relationship between single nucleotide polymorphisms of interleukin 23 receptor (IL-23R) gene and Keshan disease (KD) in Northwest Chinese Han population.Methods A total of 285 Chinese Han subjects from Huangling,Shaanxi,including 79 KD patients (case group) and 206 control subjects (control group) were involved in this study.Genomic DNA was extracted from peripheral venous blood.The polymorphism of genetic variation was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF).All sample groups were tested for Hardy-Weinberg equilibrium using goodness-of-fit x2 test.Differences in genotype distribution between two groups were compared by x2 test.Logistic regression analysis was applied to detect association using age as a confounding factor.Results The gene frequency distribution of IL-23R gene rs10889677 in case group and control group conformed to the Hardy-Weinberg equilibrium (x2 =0.254,P ＞ 0.05).Correlation analysis results:the difference of genotype frequency of IL-23R gene rs10889677 in case group (CC,CA,AA were 6.3％,36.7％,57.0％,respectively) and control group (CC,CA,AA were 5.3％,43.2％,51.5％,respectively) was not statistically significant (x2 =1.008,P ＞ 0.05).After age adjustment,there was no significant difference in genotype frequency of IL-23R gene rs10889677 (x2sdj =0.669,P ＞ 0.05) between two groups.Conclusion There is no correlation between IL-23R gene rs10889677 and KD in Northwest Chinese Han population.

Protective effects of selenium on nitric oxide mediated myocardial apoptosis / 中华地方病学杂志

Objective To investigate the protective effects of selenium on nitric oxide(NO)-mediated myocardial apoptosis.Methods The AC16 cardiomyocyte cultured in vitro were divided into control group,selenium treatmentgroup,sodium nitroprusside(SNP) treatment group and selenium + SNP treatment group,SNP was the exogenous NO donor.There was no intervention in the control group,and an equal volume of the culture solution was added to the treatment groups.The selenium treatment group added a dose of 100 μg/L of selenium,the SNP treatment group added a dose of 1.0 mmol/L of SNP,and the selenium + SNP treatment group was pretreated by 100 μg/L selenium for 4 h followed by 1.0 mmol/L SNP;the cells or supernatants were collected after 24 h of culture.The content of NO was detected by Griess method in supernatants.The level of cell reactive oxygen species was detected by flow cytometry.The changes of cell mitochondrial membrane potential and apoptosis were observed under fluorescence microscope.The real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expression levels of apoptosis-related genes B-cell lymphoma-2 (Bcl-2) associated X protein (Bax) and Bcl-2,respectively.Results The NO content in the control group,selenium treatment group,SNP treatment group and selenium + SNP treatment group were (10.3 ± 1.8),(9.2 ± 2.1),(15.2 ± 3.5),(14.3 ± 2.6) μmmol/L,respectively;SNP had a main effect on NO content (F =23.33,P ＜ 0.05).The cell reactive oxygen species were 31.63 ± 1.40,29.52 ± 2.86,60.62 ± 4.83,50.08 ± 2.41,respectively;selenium and SNP had main effects on reactive oxygen species (F =12.19,187.20,P ＜ 0.05),selenium combined with SNP had an interactive effect on reactive oxygen species (F =5.42,P ＜ 0.05).The cell mitochondrial membrane potential levels were 0.42 ± 0.11,0.37 ± 0.07,7.25 ± 1.91,and 5.21 ± 1.59,respectively;selenium and SNP had main effects on cell mitochondrial membrane potential levels (F =14.21,440.01,P ＜ 0.05),selenium combined with SNP had an interactive effect on cell mitochondrial membrane potential levels (F =12.89,P ＜ 0.05).Selenium had main effects on nuclear pyknosis ratio,Bcl-2 mRNA and Bax protein expressions (F =9.52,10.84,22.17,P ＜ 0.05);SNP had main effects on nuclear pyknosis ratio,Bax and Bcl-2 mRNA expressions,and Bcl-2 protein expression (F =192.86,21.90,16.09,18.39,P ＜ 0.05);selenium combined with SNP had an interactive effect on Bax,Bcl-2 mRNA and protein expressions (F =20.51,7.59,15.38,11.97,P ＜ 0.05).Conclusion The SNP can induce apoptosis of AC16 cardiomyocyte;selenium combined with SNP has an interactive effect on AC16 cardiomyocyte,indicating that selenium has protective effect on NO modiated myocardial apoptosis.

An association between single nucleotide polymorphisms of Bcl-2 related anti apoptotic protein 3 gene and Keshan disease in north Chinese Han population / 中华地方病学杂志

Objective To investigate the relationship between single nucleotide polymorphisms of Bcl-2 related anti apoptotic protein 3 (BAG3) gene and Keshan disease (KD) in north Chinese Han population.Methods In 2002 a total of 285 Chinese Han subjects,including 79 KD patients and 206 control subjects were involved in this study.Genomic DNA was extracted from the peripheral venous blood sample.Blood samples were provided by the Institute of Endemic Disease Prevention,Xi'an Jiaotong University,and stored at 80 ℃.The polymorphism of genetic variation was genotyped by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF).The data was analyzed using TYPER 4.0 or SPSS16.0 software.All sample groups were tested for Hardy-Weinberg equilibrium using goodness-of-fit x2 test.Differences in genotype distribution and allele frequencies between case and control were compared by x2 test.Logistic regression analysis was applied to detect association using age as a confounding factor.Results All sample group passed the Hardy-Weinberg equilibrium test (P ＞ 0.05).Significant differences were not observed in genotype distribution between cases (rs2234962:CC,CT,TT were 0.0％,0.0％ and 100.0％,respectively;rs196295:GG,GA,AA were 22.8％,54.4％ and 22.8％,respectively;rs3858339:GG,GT,TT were 5.1％,38.0％ and 56.9％,respectively;rs3858340:TT,TC,CC were 5.1％,38.0％ and 56.9％,respectively) and controls (rs2234962:CC,CT,TT were 0.0％,1.0％ and 99.0％,respectively;rs196295:GG,GA,AA were 21.4％,51.5％ and 26.2％,respectively;rs3858339:GG,GT,TT were 5.8％,34.5％ and 59.7％,respectively;rs3858340:TT,TC,CC were 5.8％,34.5％ and 59.7％,respectively) for rs2234962,rs3858339,rs196295 and rs3858340 on BAG3 gene (x2 =0.685,0.408,0.330,0.330,P ＞ 0.05).Significant differences were not observed in genotype after agecorrecting between cases and controls for 4 SNPs on BAG3 gene (x2 =0.001,0.019,1.009,0.019,P ＞ 0.05).Conclusion The results suggest that the BAG3 gene might not be a susceptibility gene of KD in north Chinese Han population.

Cellular responses to T-2 toxin and/or deoxynivalenol that induce cartilage damage are not specific to chondrocytes.

The relationship between T-2 toxin and deoxynivalenol (DON) and the risk of Kashin-Beck disease is still controversial since it is poorly known about their selectivity in cartilage damage. We aimed to compare the cytotoxicity of T-2 toxin and DON on cell lines representative of cell types encountered in vivo, including human chondrocytes (C28/I2), human hepatic epithelial cells (L-02) and human tubular epithelial cells (HK-2). In addition, we determined the distribution of T-2 toxin and DON in Sprague-Dawley (SD) rats after a single dose exposure. T-2 toxin or DON decreased proliferation in a time- and concentration-dependent manner and their combination showed a similar antagonistic effect in C28/I2, L-02 and HK-2 cells. Moreover, we observed cell cycle arrest and apoptosis, associated with increased oxidative stress and decline in mitochondrial membrane potential induced by T-2 toxin and/or DON. In vivo study showed that T-2 toxin and DON did not accumulate preferentially in the knee joint compared to liver and kidney after an acute exposure in SD rats. These results suggest that T-2 toxin and/or DON inhibit proliferation and induce apoptosis through a possible mechanism involving reactive oxygen species-mediated mitochondrial pathway that is not specific for chondrocytes in vitro or joint tissues in vivo.

Cerebral white matter lesions and silent cerebral infarcts in postmenopausal women with polycystic ovary syndrome.

OBJECTIVES: The association between polycystic ovary syndrome (PCOS) and cardiovascular disease remains unclear. Our objective was to examine the relation of PCOS with cerebrovascular disease measured by brain magnetic resonance imaging (MRI) as white matter lesions (WMLs) and silent cerebral infarcts (SCIs). METHODS: Seventy postmenopausal women with PCOS and 140 controls participated in the case-control, cross-sectional study. Clinical, anthropometric, hormonal and metabolic parameters were measured. WMLs and SCIs were evaluated by brain MRI. RESULTS: Compared with controls, postmenopausal women with PCOS had higher mean body mass index, larger waist hip ratio, higher level of low-density lipoprotein cholesterol, free androgen index and lower sex hormone-binding globulin level. The PCOS group had a significantly higher prevalence of WMLs (40% versus 22%, p = 0.009), and SCIs (17% versus 7%, p = 0.032) compared to the control group. CONCLUSIONS: Postmenopausal women with PCOS are associated with WMLs and SCIs.

Optimization of the Formulation of Azelastine Hydrochloride Thermosensitive In-situ Gel Nasal Drops by Central Composite Design-Response Surface Methodology / 中国药房

OBJECTIVE:To optimize Azelastine hydrochloride (AH) thermosensitive in-situ gel nasal drops formulation. METHODS:Using poloxamer 407(P407)and poloxamer 188(P188)as excipients,AH thermosensitive in-situ gel was prepared by cold solution method. The formulation was optimized by central composite design-response surface methodology using the amount of P407 and P188(g/100 ml)as factors and phase-transition temperature as index. Binomial expression was fitted,and pre-dicted and measured values were compared. RESULTS:The correlation coefficient R2 fitted by binomial expression was equal to 0.986 5. The optimal formulation was as follows as P407 for 20.414 4%,P188 for 5.035 4%,measured value of(30.81±0.02)℃, predicted values of 31 ℃,deviation of 0.61%. CONCLUSIONS:AH thermosensitive in-situ gel nasal drops formulation is opti-mized by central composite design-response surface methodology.

Clinical Observation of Pricking Sifeng (EX-UE10) plus Chinese Medication for Anorexia in Kids / 上海针灸杂志

Objective To observe the clinical efficacy of pricking Sifeng (EX-UE10) plus Chinese medication in treating anorexia in kids, and to provide clinical evidence for the treatment of kid’s anorexia with traditional Chinese medicine. Method Seventy-six eligible anorexia patients were randomized into an observation group and a control group. The control group was intervened by orally taking Chinese medication, while the observation group was by pricking Sifeng plus Chinese medication, and the clinical efficacies were compared 1 month later. Result After treatment, the scores of poor appetite and decreased food intake in the observation group were significantly lower than that in the control group (P<0.05); the recovery rate was 34.2% in the observation group versus 13.2% in the control group, and the recovery rate in the observation group was significantly higher than that in the control group (P<0.05). Conclusion Pricking Sifeng plus Chinese medication can produce a significant clinical efficacy in treating anorexia in kids.

Construction and identification of pcDNA3-HBsAg-p30-ROP2 expression vec-tor / 中国血吸虫病防治杂志

Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.

Application of Fourier transform profilometry in 3D-surface reconstruction / 生物医学工程学杂志

With the improvement of system frame and reconstruction methods in fluorescent molecules tomography (FMT), the FMT technology has been widely used as an important experimental tool in biomedical research. It is necessary to get the 3D-surface profile of the experimental object as the boundary constraints of FMT reconstruction algorithms. We proposed a new 3D-surface reconstruction method based on Fourier transform profilometry (FTP) method under the blue-purple light condition. The slice images were reconstructed using proper image processing methods, frequency spectrum analysis and filtering. The results of experiment showed that the method properly reconstructed the 3D-surface of objects and has the mm-level accuracy. Compared to other methods, this one is simple and fast. Besides its well-reconstructed, the proposed method could help monitor the behavior of the object during the experiment to ensure the correspondence of the imaging process. Furthermore, the method chooses blue-purple light section as its light source to avoid the interference towards fluorescence imaging.

Splenic sarcoidosis: a case report and review of literature / 中华内科杂志

Objective To describe the clinical features and imaging characteristics of nodular splenic sarcoidosis. Methods We describe a patient with splenic sarcoidosis and review the related medical literature, the etiology, symptomatology, pathology, diagnosis, differential diagnosis, management and prognosis of splenic sarcoidosis. Results The etiology of this rare disease remains unknown. Symptoms are scanty and usually mild; computed tomography usually reveals splenomegaly or the presence of multiple nodules, confusing with metastatic tumor in spleen. On histopathologic examination, sarcoid produces noncaseating granulomas. Sarcoid is typically treated only when symptomatic. Oral corticosteroids is the most important method of treatment in patients with progressive loss of organ functions. Prognosis has closed relationship with early clinical manifestation. Conclusion Splenic sarcoidosis is rare and often misdiagnosis as other diseases.

Ginsenoside Rg1 inhabits cardiomyocyte hypertrophy induced by prostaglandin F_(2?) / 中国药理学通报

Aim To study the effect of ginsenoside Rg1(Rg1)on cardio myocyte hypertrophy induced by prostaglandin F2?(PGF2?),and to probe primarily into its mechanism.Methods The cultured neonatal rat cardiomyocyte hypertrophic response and the antihypertrophic effects of Rg1 were observed by measuring the cell diameter,protein content and the expression of atrial natriuretic factor(ANF) mRNA,which was assayed by real-time PCR.For mechanism studies,the intracellular free calcium concentration(i) in cultured cardiomyocytes was measured by using Fura-2/AM as a fluorescent indicator,nitric oxide(NO) metabolite level in the culture medium was tested by using Nitric Oxide Synthase Detection Kit.Results PGF2? 0.1 ?mol?L-1 caused the increases in the cardiomyocyte cell diameter,protein content and the expression of ANF mRNA.It could increase the i in cultured cardiomyocytes.Rg1 15.6、31.2、62.4 ?mol?L-1 could concentration-dependently inhibit the cardiomyocyte hypertrophy induced by PGF2?,and the cell diameter of cardiomyocyte treated by PGF2? was decreased by 18.4%、32.7%、43.8%(P