Clinical characteristics and outcomes of 112 cardiovascular disease patients infected by 2019-nCoV / 中华心血管病杂志

Objective: To explore the clinical characteristics and prognosis of the new coronavirus 2019-nCoV patients combined with cardiovascular disease （CVD). Methods: A retrospective analysis was performed on 112 COVID-19 patients with CVD admitted to the western district of Union Hospital in Wuhan, from January 20, 2020 to February 15, 2020. They were divided into critical group (ICU, n=16) and general group (n=96) according to the severity of the disease and patients were followed up to the clinical endpoint. The observation indicators included total blood count, C-reactive protein (CRP), arterial blood gas analysis, myocardial injury markers, coagulation function, liver and kidney function, electrolyte, procalcitonin (PCT), B-type natriuretic peptide (BNP), blood lipid, pulmonary CT and pathogen detection. Results: Compared with the general group, the lymphocyte count (0.74 (0.34, 0.94)×109/L vs. 0.99 (0.71, 1.29)×109/L, P=0.03) was extremely lower in the critical group, CRP (106.98 (81.57, 135.76) mg/L vs. 34.34 (9.55,76.54) mg/L, P<0.001) and PCT (0.20 (0.15,0.48) μg/L vs. 0.11 (0.06,0.20) μg/L, P<0.001) were significantly higher in the critical group. The BMI of the critical group was significantly higher than that of the general group (25.5 (23.0, 27.5) kg/m2 vs. 22.0 (20.0, 24.0) kg/m2，P=0.003). Patients were further divided into non-survivor group (17, 15.18%) group and survivor group (95, 84.82%). Among the non-survivors, there were 88.24% (15/17) patients with BMI> 25.0 kg/m2, which was significantly higher than that of survivors (18.95% (18/95), P<0.001). Compared with the survived patients, oxygenation index (130 (102, 415) vs. 434 (410, 444), P<0.001) was significantly lower and lactic acid (1.70 (1.30, 3.00) mmol/L vs. 1.20 (1.10, 1.60) mmol/L, P<0.001) was significantly higher in the non-survivors. There was no significant difference in the proportion of ACEI/ARB medication between the critical group and the general group or between non-survivors and survivors （all P>0.05). Conclusion: COVID-19 patients combined with CVD are associated with a higher risk of mortality. Critical patients are characterized with lower lymphocyte counts. Higher BMI are more often seen in critical patients and non-survivor. ACEI/ARB use does not affect the morbidity and mortality of COVID-19 combined with CVD. Aggravating causes of death include fulminant inflammation, lactic acid accumulation and thrombotic events.

Association of smoking status with incident cardiovascular disease in the middle-aged and older male populations / 中华疾病控制杂志

Objective To investigate the association of smoking status with incident cardiovascular disease (CVD) and its subtypes among the middle-aged and older male populations. Methods This study included 13 940 males from Dongfeng-Tongji (DFTJ) cohort who were free of coronary heart disease (CHD), stroke, cancer or severely abnormal electrocardiogram (ECG) at baseline. All participants completed baseline questionnaires, physical examinations, clinical biochemical tests and blood sample collection. Cox proportional hazard models were used to estimate the hazard ratios (HRs) and 95% confident intervals (CI) for the association analyses. Results Compared with never smokers, current smokers had significant higher risks of CVD, CHD and stroke, the adjusted HRs of current smokers who smoked for more than 40 pack-years were 1.49 (95% CI: 1.32-1.68, Ptrend=0.001), 1.40 (95% CI: 1.22-1.62, Ptrend=0.026) and 1.59 (95% CI: 1.26-2.00, Ptrend=0.029) for CVD, CHD and stroke, respectively; and the adjusted HRs of current smokers who started smoking before 20 years old were 1.29 (95% CI: 1.06-1.58, Ptrend=0.007) and 1.30 (95% CI: 1.03-1.64, Ptrend=0.010) for CVD and CHD, respectively. Former smokers who had quitted smoking for 10 or more years had significant lower risks of CVD (HR: 0.80, 95% CI: 0.71-0.91, Ptrend=0.017) and stroke (HR: 0.65, 95% CI: 0.50-0.84, Ptrend=0.207) when comparing to current smokers. Conclusions Smoking is significantly associated with higher risks of CVD, CHD and stroke, and greater amount of smoking and earlier age at smoking initiation are associated with a higher risk of CVD. Smoking cessation can reduce the risk of CVD.

Analysis of prevalence and related factors of osteoporosis in retired population based on Dongfeng-Tongji Cohort / 中华预防医学杂志

Objective@#To investigate the prevalence and related factors of osteoporosis among retired population in Dongfeng-Tongji cohort.@*Methods@#27 009 retired participants were recruited from Dongfeng Motor Corporation in Hubei Province in 2008 and followed up from April to October in 2013. newly retired participants also were recruited. Data were collected by using questionnaire, physical examination, serum hepatase detection and bone densitometry. Totally, 30 916 participants were included for data analysis after excluding participants with severe bone metabolic diseases, taking hormone drugs, incomplete follow-up data and who were under 45 years old. Age-standardized prevalence of osteoporosis was calculated according to data of the 2010 Sixth National Population Census. Multivariate logistic regression analysis was applied to explore the associated factors of osteoporosis.@*Results@#Prevalence of osteoporosis was 42.3% (13 083/30 916) and age standardized prevalence was 40.7%: 35.0% (4 854/13 878) and 34.8% for males; 48.3% (8 229/17 038) and 47.1% for females. Significantly associated factors with osteoporosis for both males and females included: older age (male: OR=1.67, 95%CI: 1.40-1.99; female:OR=3.34, 95%CI: 2.70-4.13), lower BMI (male: OR=1.70, 95%CI: 1.40-2.06; female: OR=1.27, 95%CI: 1.04-1.53), exercise (male: OR=0.69, 95%CI: 0.61-0.78; female: OR=0.87, 95%CI: 0.80-0.96), abnormal elevated serum alkaline phosphatase (ALP) (male: OR=1.12, 95%CI: 1.01-1.24; female: OR=1.15, 95%CI: 1.06-1.25), γ-glutamyltransferase (γ-GT) (male: OR=1.16, 95%CI: 1.02-1.30; female: OR=1.13, 95%CI: 1.03-1.24) and aspartate transaminase/alanine aminotransferase (AST/ALT) (male: OR=1.15, 95%CI: 1.05-1.25; female: OR=1.28, 95%CI: 1.19-1.38). Smoking (OR=1.27, 95%CI: 1.07-1.39) and drinking (OR=1.11, 95%CI: 1.08-1.16) were associated factors for males while menopausal (OR=1.67, 95%CI: 1.47-1.89) for females. There were positive dose-response correlation relationships of serum levels of ALP, γ-GT and AST/ALT with osteoporosis (all P values<0.05).@*Conclusion@#Osteoporosis was relatively common among retired population in Dongfeng-Tongji cohort. In addition to known factors such as older age, lower BMI and exercise, abnormal elevated serum ALP, γ-GT and AST/ALT were also associated with osteoporosis.

Association of alcohol use and incidence of type 2 diabetes mellitus in the middle-aged and elderly male population: a prospective cohort study / 中华预防医学杂志

Objective@#To investigate the association between alcohol use and incidence of type 2 diabetes mellitus (T2DM) in the middle-aged and elderly male population.@*Methods@#All participants were from Dongfeng-Tongji cohort, 27 009 retired employees from Dongfeng Motor Corporation in Hubei Province were enrolled in the Dongfeng-Tongji cohort baseline survey in 2008. In baseline study, information of alcohol use and other covariates were collected by semi-structured questionnaire and all participants completed physical examination including the test of fasting glucose and blood lipid levels. A total of 6 784 male participants from Dongfeng-Tongji cohort who were without diagnosis of diabetes, coronary heart disease, stroke, or cancer in baseline study were enrolled in this study. We completed the first follow-up in 2013 and the outcome of disease or death was retrieved based on health-care medical records according to the unique medical insurance ID. Cox proportional hazard regression model was used to estimate the association between alcohol use and incidence of type 2 diabetes mellitus (T2DM), by drinking features and patterns.@*Results@#Out of the 6 784 participants, 3 541 participant were defined as non-alcohol drinkers and there were 15 852.2 person-years of follow-up; among which 270 new cases of T2DM were diagnosed withthe crude incidence density of non-alcohol drinkers at 1 703.2/100 000 person-years. The other 3 243 subjects were classified as alcohol drinkers and there were 14 509.8 person-years of follow-up; and among which 258 new cases of T2DM were diagnosed, with the crude incidence density of T2DM at 1 778.1/100 000 person-years. Multivariate COX proportional hazard regression model indicated that there was no significantly increased risk of T2DM incidence between alcohol drinkers and non-alcohol drinkers(HR(95% CI): 1.09 (0.91- 1.30)). However, participants who averagely consumed >20 g/d or>7 times/week had a significantly increased risk of T2DM compared with non-alcohol drinkers, and the value of HR(95%CI) was 1.27 (1.02- 1.58) and 1.35 (1.00- 1.83), respectively. Among men who consumed alcoholic beverages more than 7 times/week, HR (95%CI) for T2DM incidence in the subjects who consumed 0.01 to 40 g and > 40 g once a time were 1.48 (1.05- 2.09) and 1.27 (0.80- 2.10), respectively.@*Conclusion@#Although we found no relationship between alcohol use and T2DM incidence overall, alcohol use more than 20 g/d or more than 7 times/week would increase the risk of T2DM.

Association between smoking and dopamine receptor D2 polymorphisms in male patients with schizophrenia / 中国心理卫生杂志

Objective:To investigate the association between dopamine receptor D2(DRD2) polymorphisms and smoking in male patients with schizophrenia.Methods:Totally 773 patients with schizophrenia (567 smokers and 206 non-smokers) and 302 normal controls (168 smokers and 134 non-smokers) were recruited.The two single nucleotide polymorphisms (SNPs) (rs1800497 and rs1079597) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP).SHEsis genetic analysis platform was used to calculate linkage disequilibrium index and infer allele distribution and haplotype frequency.Results:There was no significant difference in two SNPs genotype and allele distributions between the patients and normal controls or between smokers and non-smokers in either patients or normal controls alone (Ps ＞ 0.05);the frequency estimations of haplotype C-A and T-G in patients with schizophrenia were higher than in normal controls (8.0％ vs.5.2％,10.2％ vs.4.1％,Ps ＜0.05),T-A (34.6％ vs.40.2％,P ＜0.05),whereas the frequency estimation of haplotype T-A in patients with schizophrenia was lower than in normal controls,and all the differences were statistically significant (34.6％ vs.40.2％,P ＜ 0.05).It was also observed that the frequency estimation of haplotype T-A in normal smokers was significantly lower than in normal non-smokers (2.5％ vs.6.1％,P ＜0.05).Conclusion:There may be a correlation between DRD2 polymorphisms and the susceptibility to schizophrenia,but not between DRD2 polymorphisms and smoking neither in patients with schizophrenia nor in normal controls.

Elevated Expression of CD93 in Serous Exosomes from Patients with Cryptococcal Meningitis and Its Significance / 现代检验医学杂志

Objective To examine the expression of CD93 in serous exosomes from patients with cryptococcal meningitis,fur-ther explore its clinical significance.Methods The 38 experimental serum samples were from patients who received the diag-nosis with cryptococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from November 2012 to De-cember 2016.The diagnosis standardization was that the cerebrospinal fluid dyeing was positive or the culturing was posi-tive.The 38 controls were collected from the health individuals examined at the same time.The exosomes in serum was sepa-rated by the ultracentrifuge method.Magnetic bead-capture combined with the flow cytometry method was used to identify antigens on the surface of exosomes.The protein level of cytokines in serum was quantified by ELISA method.The compari-son of tested values from experimental and controlled groups was measured by two independent samples't test,and the cor-relation between two variates was showed by Pearson coefficient.Results The results of flow cytometry showed that the ex-pression of CD93 on exosomes in the serum samples from experimental group was higher than that in the controlled group, which was(79.11±19.31 vs 23.98±6.56)%,with difference in statistics(t=16.66,P<0.000 1).The expression levels of IFN-γin serum samplefrom experimental and control groups were(39.78±10.77 vs 58.98±16.99)pg/ml,with differ-ence in statistics(t=5.884,P<0.000 1).The expression levels of IL-17 in serum sample from experimental and control groups were(16.32±4.03 vs 3.11±0.87)pg/ml,with difference in statistics(t=19.75,P<0.000 1).The expression lev-els of IL-1β in serum sample from experimental and control groups were(57.12 ± 12.98 vs 13.45 ± 4.78)pg/ml,with difference in statistics(t=19.46,P<0.000 1).The percentage of CD93+exosomes was positively correlative with IFN-γ, IL-17,IL-1β(r=0.488,0.456,0.532,P<0.01)in serum samples from the experimental group and there was statistical difference.Conclusion In the disease process of cryptococcal meningitis,the expression of CD93 on exosomes might involve in the diseases'processes by affecting the CD4+T cells.

Protective effects of Cyclopamine in adjuvant arthritis rats kidney injury and mechanism research / 中国免疫学杂志

Objective:Observed activity of adjuvant arthritis ( AA) rats Hedgehog signaling pathway and explore the effect of Cyclopamine on rat arthritis and kidney injury.Methods:40 rats were randomly divided into control group,Cyclopamine group,AA+Cyclopamine group and AA model group.We used Freund′s complete adjuvant rat model with Cyclopamine intraperitoneal injection.By measuring paw swelling, systemic inflammation and arthritis semi-quantitative assessment methods of rats to evaluate the model.HE staining was used to detect the renal pathological changes of rats.Western blot was used to detect kidney Gli1 protein expression levels of the rats.Immunohistochemical staining was used to detect the rat kidney TNF-α,IFN-γ,IL-6 expression.Results:After using Cyclo-pamine,the AA rat paw swelling reduced and arthritis refers relieved significantly.Compared with the control group,Cr,BUN and organ coefficient increased significantly in AA model group, and the difference was statistically significant.Meanwhile renal TEM detection appeared obvious pathological changes in rats of AA model group.After using cyclopamine,the content of Cr,BUN and organ coefficient change reduced significantly, so did the pathological.Western blot detected kidney tissue Gli1 protein in each group.Compared to control group,there was no significant difference in Cyclopamine group.AA model group and AA+Cyclopamine group Gli1 protein expression was significantly higher compared with Cyclopamine Gli1 protein group,the difference was statistically significant.AA model group and AA Cyclopamine group Gli1 protein expression was significantly higher,the difference was statistically significant.Compared to the AA model group,AA+Cyclopamine group Gli1 protein levels have decreased significantly,the difference was statistically signifi-cant.Immunohistochemical assay detection found that kidney tissue proinflammatory cytokines TNF-α,IFN-γ,IL-6 expression and semi-quantitative score changed.Compared with the control group, TNF-α, IFN-γexpression of AA model group were significantly increased.After using Cyclopamine,the expression of TNF-α,IFN-γin kidney of AA model reduced significantly.IL-6 expression in AA model group was significantly higher than the control group.Conclusion:Cyclopamine AA can relieve arthritis and kidney injury in rats with arthritis AA,the Hh pathway was on activity state in the process,may altered expression of inflammatory factors.

Effects and clinical significance of pentoxifylline on the oxidative stress of rats with diabetic nephropathy / 华中科技大学学报（医学）（英德文版）

Diabetic nephropathy (DN) is a common and serious clinical complication of diabetes and presently there are no effective ways to prevent its occurrence and progression. Recent studies show that pentoxifylline (PTX) can improve renal hemodynamics, reduce urinary protein excretion, and alleviate or delay renal failure in DN patients. In this study, we focused on the anti-oxidative stress effect of PTX on alleviating renal damages of DN using rat models. DN rats were established with injection of streptozotocin. Blood glucose, urinary protein excretion, serum cystatin C, renal biopsy, superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and renal homogenate and renal nitrotyrosine levels were analyzed before and 12 weeks after the treatment of PTX. Before treatment, all the DN rats had elevated blood glucose, increased urinary protein excretion and elevated serum cystatin C. Morphologically, DN rats exhibited renal tissue damages, including swelling and fusions of foot processes of podocytes under electron microscope. Masson staining revealed blue collagen deposition in glomeruli and renal interstitium. With treatment of PTX, symptoms and renal pathological changes of DN rats were alleviated. Furthermore, the MDA levels were increased and the SOD levels were decreased in the serum and kidneys of DN rats, and these changes were reversed by PTX. The expression of nitrotyrosine was up-regulated in DN rat model and down-regulated by PTX, indicating that PTX was able to inhibit oxidative reactions in DN rats. PTX could alleviate renal damage in DN, which may be attributable to its anti-oxidative stress activity.

Effects and clinical significance of pentoxifylline on the oxidative stress of rats with diabetic nephropathy / 华中科技大学学报（医学）（英德文版）

Diabetic nephropathy (DN) is a common and serious clinical complication of diabetes and presently there are no effective ways to prevent its occurrence and progression. Recent studies show that pentoxifylline (PTX) can improve renal hemodynamics, reduce urinary protein excretion, and alleviate or delay renal failure in DN patients. In this study, we focused on the anti-oxidative stress effect of PTX on alleviating renal damages of DN using rat models. DN rats were established with injection of streptozotocin. Blood glucose, urinary protein excretion, serum cystatin C, renal biopsy, superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and renal homogenate and renal nitrotyrosine levels were analyzed before and 12 weeks after the treatment of PTX. Before treatment, all the DN rats had elevated blood glucose, increased urinary protein excretion and elevated serum cystatin C. Morphologically, DN rats exhibited renal tissue damages, including swelling and fusions of foot processes of podocytes under electron microscope. Masson staining revealed blue collagen deposition in glomeruli and renal interstitium. With treatment of PTX, symptoms and renal pathological changes of DN rats were alleviated. Furthermore, the MDA levels were increased and the SOD levels were decreased in the serum and kidneys of DN rats, and these changes were reversed by PTX. The expression of nitrotyrosine was up-regulated in DN rat model and down-regulated by PTX, indicating that PTX was able to inhibit oxidative reactions in DN rats. PTX could alleviate renal damage in DN, which may be attributable to its anti-oxidative stress activity.

Using the stable HSPA1A promoter-driven luciferase reporter HepG2 cells to assess the overall toxicity of coke oven emissions / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>Using the stable HSPA1A (HSP70-1) promoter-driven luciferase reporter HepG2 cells (HepG2/HSPA1A cells) to assess the overall toxicity of coke oven emissions.</p><p><b>METHODS</b>The stable HepG2/HSPA1A cells were treated with different concentrations of coke oven emissions (COEs) collected from the top, side, and bottom of a coke oven battery for 24 h. After the treatments, luciferase activity, cell viability, malondialdehyde (MDA) concentration, Olive tail moment, and micronuclei frequency were determined, respectively.</p><p><b>RESULTS</b>The bottom COEs induced significant increases (P < 0.01) in relative luciferase activity up to 1.4 times the control level at 0.15 µg/L. The low dose of side COEs (0.02 µg/L) led to a significant increase (P < 0.01) in relative luciferase activity that progressively increased to 2.1 times the control level at 65.4 µg/L. The top COEs produced a strong dose-dependent induction of relative luciferase activity up to over 5 times the control level at the highest concentration tested (202 µg/L). In HepG2/HSPA1A cells treated with the bottom COEs, relative luciferase activity was positively correlated with MDA concentration (r = 0.404, P < 0.05). For the three COEs samples, positive correlations were observed between relative luciferase activity and Olive tail moment and micronuclei frequency.</p><p><b>CONCLUSION</b>The relative luciferase activity in HepG2/HSPA1A cells can sensitively reflect the overall toxicity of COEs. The stable HepG2/HSPA1A cells can be used for rapid screening of the overall toxicity of complex air pollutants in the workplace.</p>

Rapid cloning and functional characterization of hypericin synthase gene / 药学学报

Hypericin, a red-colored naphtodianthrone, is a natural product synthesized in the medicinal plant Hypericum perforatum, commonly known as St. John's wort. Hypericin has attracted a growing attention of the pharmaceutical industry because of its potential application to various therapies, including the treatment of depression and remarkable antiviral and photodynamic activities, hyp-1 gene encodes for phenolic coupling protein which catalyzes in vitro direct and specific conversion of emodin to hypericin which, however, has not formed common opinion so far. Six pairs of primers specific to hyp-1 gene were synthesized. The rapid cloning of hyp-1 gene was performed based on step-by-step extension of a short region of the gene through a series of PCR reactions. All cloned sequences were confirmed by DNA sequencing. A vector named pET32ahyp containing hyp-1 gene was constructed and was transformed into E. coli to induce heterologous expression. SDS-PAGE and Western blot results showed the recombinant Hyp-1 protein was expressed successfully in E. coli. The soluble fraction was used to test the function of the recombinant Hyp-1. Hypericin was detected by LC-MS/MS with emodin as a substrate under in vitro conditions. The above results corroborated the Hyp-1 function, a confusing question, which lay a material foundation for the synthesis of hypericin by synthetic biotechnology.

Molecular genetics of globozoospermia: an update / 中华男科学杂志

Globozoospermia, as a severe teratozoospermia caused by gene mutations, is a rare congenital disease with main clinical manifestations of the round head of sperm and abnormality or absence of acrosome, and its precise mechanism is not yet clear. Studies show that the pathogenic genes associated with globozoospermia include SPATA16, PICK1, GOPC, Hrb, Csnk2a2 and bs. This paper outlines the progress in the studies of molecular genetics of globozoospermia, aiming to contribute to the molecular diagnosis and mechanism investigation of the disease.

Flow cytometry for evaluation of sperm plasma membrane integrity in varicocele patients / 中华男科学杂志

<p><b>OBJECTIVE</b>To detect the sperm plasma membrane integrity (PMI) of varicocele (VC) patients using SYBR-14/PI fluorescent staining and flow cytometry, and investigate its clinical significance.</p><p><b>METHODS</b>We collected semen samples from 120 men, including 30 grade-1 varicocele patients (VC1), 30 grade-2 (VC2), 30 grade-3 (VC3), and 30 normal fertile volunteer controls. Conventional semen analyses were performed by computer-assisted semen analysis (CASA). All the semen samples were washed with PBS and then subjected to SYBR-14/PI staining for the detection of sperm PMI by flow cytometry. The proportion of normal sperm with PMI was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI- %), sperm PMI was determined and sperm fertilization capacity predicted.</p><p><b>RESULTS</b>Significant differences were detected in SYBR-14+/PI- and SYBR-14-/PI+ between the normal men and varicocele male patients (P < 0.01). The percentages of the sperm with PMI (SYBR-14+/PI- %) were remarkably lower in the VC1, VC2 and VC3 groups ([54.85 +/- 3.78]%, [45.37 +/- 4.12]% and [35.14 +/- 4.91]%) than in the normal controls ([70.79 +/- 6.71]%). SYBR-14+/PI-% was correlated positively with sperm motility (r=0.965, P < 0.01) and the percentage of grade a + b sperm (r = 0.874, P < 0.01), negatively with the percentage of grade d sperm (r = -0.965, P <0.01), but not significantly with pH, semen volume and liquefaction time (P > 0.05).</p><p><b>CONCLUSION</b>SYBR-14/PI fluorescent staining and flow cytometry can quickly and exactly detect sperm PMI. Varicocele decreases sperm PMI, which might be an important cause of male infertility.</p>

Ureaplasma urealyticum infection affects sperm plasma membrane integrity in infertile men / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the impact of Ureaplasma urealyticum (Uu) infection on the integrity of sperm plasma membrane in infertile males.</p><p><b>METHODS</b>Sixty-three semen samples were divided into a Uu infection group (n = 32) and a normal control group (n = 31). Conventional semen analyses were performed by computer-assisted semen analysis (CASA) and Uu detected by the culture method. The semen samples were washed with PBS and dyed by SYBR-14/PI double fluorescent staining, followed by detection of the integrity of sperm plasma membrane by flow cytometry. The percentage of the sperm with intact plasma membrane was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI-%).</p><p><b>RESULTS</b>The Uu infection group showed a significantly decreased integrity of sperm plasma membrane ([45.14 +/- 10.69]%) and reduced percentage of grade a + b sperm ([23.29 +/- 8.81]%) as compared with the normal control group ([72.68 +/- 9.91]% and [46.32 +/- 9.54]%) (P < 0.01). But there were no significant differences in the semen volume, pH value, and sperm concentration between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>Uu infection decreases the integrity of sperm plasma membrane, which might be an important factor of male infertility.</p>

The role of miRNA-122 expression during the acute liver failure in mice induced by D-GalN/LPS / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression of miR-122 and its relationship with progression and development of acute liver failure in mice induced by D-GalN/LPS, and to explore new biomarker(s) for early diagnosis of acute liver failure.</p><p><b>METHODS</b>BALB/C mice were randomly divided into four groups: the mice were given D-GalN (900 mg/kg body weight) and LPS (10 micog/kg body weight) intraperitoneally (i.p.) to construct the acute liver model; whereas the control groups were given D-GalN (900 mg/kg), LPS (10 microg/kg) and normal saline respectively. All biochemical and histological indexes were determined at 0, 1, 3, 5, 7 and 9 h respectively after administration. Real-time RT-PCR were used to detect the expression of miR-122 and pro-inflammatory cytokines, furthermore, the expression of miR-122 was verified by LNA (lock nucleic acid)-Northern-blot. ALT and AST levels were tested by biochemistry analyzer. Serum pro-inflammatory cytokine levels were tested by ELISA.</p><p><b>RESULTS</b>The mortality rate was about 80% at 24h after D-GalN/LPS treatment, but no mortality was observed in the other three control groups. Liver special miRNA miR-122 was highly expressed in liver tissue of normal mice (ct is approximately equal to 14), it was up-regulated significantly (P = 0.013) at first hour after treatment then down-regulated according to the development of acute liver failure, the change was more obvious at 9 h (ct is approximately equal to 15, P = 0.002). ALT and AST levels increased obviously at 3h after treatment and reached peak at 7 hours then they were declined sharply. It was found that the expression of miR-122 was faster and more durable than ALT. Pro-inflammatory cytokines related to acute liver failure including TNFa and IL-6 were all up-regulated in serum as well as liver tissue (P less than 0.05). Correlation analysis showed that miR-122 had a negative correlation with ALT (correlation coefficients -0.505) and positive correlations with TNFa and IL-6 (correlation coefficients were 0.493 and 0.674 respectively).</p><p><b>CONCLUSIONS</b>Liver-specific miR-122 supposed be a new marker molecule for early diagnosis of liver cells injury in the acute liver failure.</p>

Study on the association of -689C/T polymorphism in the PPARgamma2 promoter with myocardial infarction / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of -689C/T polymorphism in the peroxisome proliferator-activated receptor-gamma2 (PPARgamma2) promoter with myocardial infarction (MI).</p><p><b>METHODS</b>This is a case-control study, which included 194 subjects with MI and 693 subjects without MI in nondiabetic Han population in Wuhan. Polymerase chain reaction-restriction fragment length polymorphism was used to determine the -689C-->T substitution.</p><p><b>RESULTS</b>The CC,CT, and TT genotype frequencies of -689C/T polymorphism were 88.1%,11.9%,and 0.0 in MI patients and 93.1%,6.6%,and 0.3% in controls, respectively (CC vs. CT+TT, P=0.025). The -689T allele was an independent risk factor for MI (OR=2.125, 95%CI: 1.206-3.744, P=0.009) after adjusting for age,sex,waist circumference,body mass index, smoking, alcohol drinking, physical activities, systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, triglyceride, level. The -689T allele carriers had significantly higher TC levels than noncarriers [(5.05+/-1.16) mmol/L vs. (4.78+/-1.05) mmol/L, P=0.004] in the total population.</p><p><b>CONCLUSION</b>The PPARgamma2 promoter -689C/T polymorphism is associated with an increased risk of MI.</p>

Dot-immunobinding assay for the activity of egg yolk anibody / 中国组织工程研究

BACKGROUND: There are differences in physical and biological activity between the antibody from mammals and egg yolk antibody (IgY) from chicken. IgY is acid- and heat-resistant, and can prevent and cure the infectious diseases in animals and human being, which is also benefit to develop routine diagnostic immunoassays. Conventional ELISA assay for IgY takes much more time than dot-immunobinding assay.OBJECTIVE: To detect the IgY stability byusing dot-immunobinding assay.DESIGN: Open trail.SETTING: Department of Transfusion, Kunming General Hospital of Chinese PLA.MATERTALS: The experiment was completed in the Kunming General Hospital of Chengdu Military Area Command of Chinese PLA from January to June 2006. Two White Leghorn hens (30 weeks old) were selected. HLA-A*0201 α chain served as the antigen. The total protein concentration of the purified antigen was 0.04 g/L with the molecular mass of 32 000(self-prepared); nitrocellulose filter (NC, import and divided); nonfat dry milk (Anyi Corp. No. 20051220); DAB (Boshide Corp.);caprylic acid (made by Shanghai Xinghuo Chemical Factory); ammonium sulfate (Shantou Guanghua Chemical product).METHODS: ①HLA-A*0201 α chain with the total protein concentration of 0.04 g/L was purified with egg yolk antibody,and identified by SDS-PAGE. ②1 μL antigen was spotted into the center of NC membrane and dried in the incubator at 37 ℃. Then the NC membrane was blocked in 1 mL PBST and put in the incubator at 37℃ with shaking in 90 r per minute for 15 minutes. Then the liquid was exchanged with 1 mL PBST and added the primary antibody at a final concentration of 10 mg/L. After 30 minutes shaking in the incubator at 37 ℃, the NC membrane was washed in PBST for three times. The second antibody, mouse anti-chicken IgY conjugated to horseradish peroxidase (HRP) was added and after 30 minutes incubation, the NC membrane was washed three times in PBST. Binding was revealed by incubation with a DAB reagent. A positive reaction was represented by adeep brown spot,Irdlcating that IgY had better activity; if the spot became lighter IgY lost part activity, and when the spot disappeared, the IgY lost a the activty.According to intensity (gray degree)of the dot compared tothe standard, the remained percent of activity of the IgY was calculated. ③IgY was adjusted to three different protein concentrations with PBS: 1, 0.1, 0.01 g/L and stayed at room temperature for four months. 10 μg lgY was taken out from each concentration sample every month to detect the activity by dot-immunobinding assay. ④IgY was put into seven EP tubes with 100 μL per tube and numbered 1-7. Number 1 to 3 was adjusted pH to 5, 3 and 2, respectively with 1 mol/L HCI; Number 4 to 6 was to 9, 11 and 12, respectively with 1 mol/L NaOH. The pH of number 7 was neutral without adding acid or base. The samples were stayed in incubator at 37 ℃ for 3 hours. 10 μg IgY from each tube was taken every hour to detect the stability at different pH by dot-immunobinding assay. ⑤IgY was added to six EP tubes (10 μL per tube) and numbered 1-6. Number 1-6 was put into waterbath at 30, 40, 50, 60, 70 and 80 ℃ for 15 minutes. After cooled in refrigerator at 4 ℃, 10 mg samples from each tube and standard sample (untreated sample) taken to check the thermal stability by dot-immunobinding assay.MAIN OUTCOME MEASURES: ①SDS-PAGE of IgY. ②IgY stability at room temperature. ③IgY stability at different pH. ④ Detection of IgY thermal stability.RESULTS: ①Purified IgY after SDS-PAGE had two major binds, the molecular mass of the heavy chain was 66.000,and the light chain was 25 000. ②1, 0.1, 0.01 g/L IgY still had partial activity after staying at room temperature for four months. ③When pH ranged from 5 to 9, IgY still had partial activity after staying in 37 ℃ for 3 hours. If pH was lower than 5 or higher than 9, it lost the whole activity in above condition. ④Purified IgY was added to six EP tubes, the number 1-4 still had partial activity, but number 5 and 6 showed some white precipitate, which was caused by protein denaturation at higher temperature.CONCLUSION: IgY stability is higher than others. The dot-immunobinding assay described a rapid and simple method for the demonstration and characterization of functional activity of egg yolk antibody. With only small volume antigen and antibody, and specific dot, the dot-immunobinding assay method could process many samples at the same time.

Application of low dose of fludarabine and cyclophosphamide combined with donor NK Cells as a non-myeloablative conditioning regimen for the haploidentical hematopoietic stem cell transplantation in mice / 中国实验血液学杂志

This study was aimed to investigate the feasibility of low dose of fludarabine, cyclophosphamide combined with donor derived alloreactive NK cells as a new nonmyeloablative conditioning regimen in the haploidentical hematopoietic stem cell transplantation (haploidentical HSCT). F1 derived-NK cells were enriched with MACS magnetic separation system, in which the proportions of the Ly49C+ and Ly49A+ cells were detected by flow cytometry and the alloreactivity was measured by LDH method. The haploidentical HSCT models were constructed, and the myeloablativity in vivo, donor engraftment and the intensity of GVHD were compared between different myeloablative and nonmyeloablative conditioning regimens, including 9 Gy TBI, 6.5 Gy TBI, flu + cy, and flu + cy + allo-NK. The results showed that the flu + cy + allo-NK conditioning was nonmyeloablative, but the rate of donor chimerism after haploidentical HSCT was significantly higher as compared with other nonmyeloablative methods, which were (28.70 +/- 5.90)% in bone marrow and (46.40 +/- 5.00)% in spleen at day 21 post-transplantation. When compared with the flu + cy conditioning, the intensity of GVHD was slight in the flu + cy + allo-NK group, in which only a half of C57BL/6 recipients experienced weight loss, and no distinct pathological damages observed in the liver, intestine, kidney and skin samples. It is concluded donor derived-alloreactive NK cells can facilitate engraftment of the haploidentical hematopoietic stem cells and mitigate GVHD. The flu + cy + allo-NK conditioning provides a new method for those elder patients with high-risk solid tumor undergoing haploidentical-HSCT.

Polymorphisms of hsp 60 gene in Chinese Han people / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To identify the single nucleotide polymorphisms (SNPs) in the regulatory and coding regions of heat shock protein 60 gene and search for its genetic makers in Chinese Han people.</p><p><b>METHODS</b>The 5' flank region, parts of the exons and introns of hsp60 gene were resequenced to identify the SNPs in Chinese Han people, and then the sequenced results to the Japanese, European and African's data in National Center for Biotechnology Information (NCBI) and HapMap databases were compared.</p><p><b>RESULTS</b>One novel SNP was identified in exon 2 resulting in synonymous variant and the G allele frequency was 0.025. There were 11 reported SNPs in the sequenced region. The minor allele frequencies of rs1116734, rs3749095, rs1050347, rs8539 were 0.51, 0.30, 0.29, 0.49. The heterozygosity of the other 7 SNPs was 0. The distributions of rs1116734, rs1050347, rs8539, rs3749095 in Chinese Han people were similar to the Japanese's. The hsp60 rs3749095 which was not found in Japanese people was a high-frequency SNP in Chinese Han people; the distribution of rs8539 in Chinese Han people was quite different from European and African's (P < 0.01).</p><p><b>CONCLUSION</b>The SNPs of hsp60 in Chinese Han people are different from the other peoples. The SNPs of hsp60 gene rs1116734, rs3749095, rs1050347, rs8539 are very common in Chinese Han people and might be used for candidate genetic markers of hsp60 gene.</p>

Glutathione S-transferase M1, T1 genotypes and the risk of mountain sickness / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the relationship between genetic polymorphisms of glutathione S-transferase (GST) M1, T1 and susceptibility to mountain sickness.</p><p><b>METHODS</b>Forty-three soldiers with acute mountain sickness and 80 healthy soldiers matching with sex/age and training under the same condition were divided into case group and control group. A multiple polymerase chain reaction method was used to detect GSTM1 and GSTT1 genes in genomic DNA isolated from peripheral blood cells from both cases and controls.</p><p><b>RESULTS</b>The frequency of the GSTT1 positive genotype was significantly higher in cases (69.8%) than in controls (42.5%) (P = 0.004, OR = 3.12, 95% CI 1.42 approximately 6.86). The frequency of GSTM1 negative genotype was also higher in cases (72.1%) than in controls (52.5%) (P = 0.03, OR = 2.34, 95% CI 1.05 approximately 5.02). Persons with both GSTM1 and GSTT1 negative genotypes had 5-fold more risk than those with GSTT1 negative and GSTM1 positive genotypes in developing mountain sickness (OR = 5.04, 95% CI: 1.00 approximately 25.3).</p><p><b>CONCLUSION</b>Genetic polymorphisms of glutathione S-transferase M1, T1 may be the risk factors in the development of mountain sickness.</p>