[Risk stratification and low-density lipoprotein cholesterol goal attainment rates in patients with very high-risk or extreme high-risk atherosclerotic cardiovascular diseases regarding three guidelines].

Objective: To explore the differences of risk stratification of very high-risk or extreme high-risk atherosclerotic cardiovascular diseases (ASCVD) and the attainment rates of low-density lipoprotein cholesterol (LDL-C) management targets evaluated by three different criteria, and the causal attributions of these differences. Methods: Patients with ASCVD were consecutively enrolled from January 1 to December 31 in 2019, and were evaluated for very high-risk or extreme high-risk and LDL-C goal attainment rates with 2018 American guideline on the management of blood cholesterol (2018AG), 2019 China Cholesterol Education Program (CCEP) Expert Advice for the management of dyslipidemias (2019EA) and 2020 Chinese expert consensus on lipid management of very high-risk ASCVD patients(2020EC), respectively. The causal attributions of the differences in attainment rates were analyzed as well. Results: A total of 1 864 ASCVD patients were included in this study. According to 2018AG, 2019EA and 2020EC, the proportions of the patients with very high-risk or extreme high-risk were 59.4%, 90.7%, and 65.6%, respectively. The absolute LDL-C target attainment rates were 37.2%, 15.7%, and 13.7%, respectively, the differences between each two rates were statistically significant (all P<0.001). As to the differences in attainment rates between 2020EC and 2018AG, 61.5% were due to the different LDL-C goal attainment values and 38.5% were caused by the different risk stratifications, while for the differences between 2020EC and 2019EA attainment rates, different LDL-C goal attainment values were responsible for 13.2%, and different risk stratifications were responsible for 86.8% of the differences. Conclusions: There are significant differences in the proportions and LDL-C attainment rates among the three different criteria for very high-risk or extreme high-risk ASCVD. 2020EC showed a moderate proportion of patients with extreme high-risk, and had the lowest LDL-C attainment rate. The differences between 2020EC and 2018AG are mainly due to the LDL-C target values, and the differences between 2020EC and 2019EA are mainly caused by the risk stratifications.

[Clinical characteristics and prognosis of young (<35 years) patients with acute ST-segment elevation myocardial infarction].

Objective: To analyze the risk factors and clinical characteristics as well as long-term prognosis of young patients (aged 18-35 years) with acute ST-segment elevation myocardial infarction (STEMI). Methods: In this retrospective study, from January 2007 to December 2017, STEMI patients who were hospitalized in Beijing Anzhen Hospital, Capital Medical University and younger than 45 years old were collected. Patients were divided to the 18-35 years old group and 36-44 years old group. The basic information of patients, clinical laboratory results, surgical information and discharge diagnosis of enrolled patients were extracted from the electronic medical record system. Subgroup analysis on STEMI patients aged 18 to 35 years was performed to compare the clinical features and outcome of patients with normal coronary angiography or stenotic coronary angiography. Results: 496 patients (20.3%) were between 18 and 35 years old, 480 cases (96.8%) were men, and 371 cases (74.8%) were smokers. The proportion of hypertension and diabetes was lower, but percent of obese (122 cases (43.3%)), level of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), uric acid and homocysteine ​​(Hcy) were significantly higher in patients aged 18 to 35 years compared with STEMI patients aged 36 to 44 years (all P<0.05). In the 18-35 years old group, there were 53 patients (10.7%) with normal coronary angiography and 443 patients (89.3%) with stenosis. The age, proportion of hypertension and diabetes, TC, LDL-C, and triglyceride (TG) levels were lower in the normal coronary angiography group than those in the coronary artery stenosis group (all P<0.05). The main coronary artery lesions were single vessel lesions (263 cases (59.4%)), and the main culprit vessels were left anterior descending artery lesions (238 cases (53.7%)). The follow-up time was 7.0 (4.0, 10.0) years, cardiovascular events were reported in 62 patients (18.9%), of which 14 patients (3.2%) died. The survival rate of patients without cardiovascular events in normal coronary angiography group was higher than that in stenosis group (P=0.029). Multivariate Cox regression analysis showed that diabetes (HR=2.713, 95%CI 1.479-4.976, P=0.001) and dyslipidemia (HR=2.819, 95%CI 1.564-5.079, P=0.001) were independent risk factors for recurrence of cardiovascular events in adult STEMI patients aged 18 to 35 years. Conclusions: STEMI patients aged 18 to 35 years were featured by male sex, obese and smokers. The proportion of hypertension and diabetes was low, while the levels of LDL-C, uric acid and Hcy were high in these patients. Coronary artery stenosis was common, and the stenosis was more likely to occur in the left anterior descending branch. Patients with normal coronary angiography had a better prognosis than those with stenosis. Diabetes and hyperlipidemia increased the risk of recurrent cardiovascular events.

[Difference on prevalence and trends of conventional risk factors for ASCVD between young Chinese and American adults with first acute myocardial infarction].

Objective: This study aimed to compare the prevalence and trends of conventional risk factors for atherosclerotic cardiovascular disease(ASCVD) between young Chinese and American adults with first acute myocardial infarction. Methods: This was a retrospective cohort analysis. Hospitalized yang adults (aged from 18 to 44 years old) with first acute myocardial infarction(AMI) from January 2007 through December 2017 were identified from Beijing Anzhen hospital medical record system. Prevalence and trends of hypertension, diabetes mellitus, obesity, smoking, and dyslipidemia were analyzed and compared with young American adults, whose data were reported by Yandrapalli et al, and the hospitalizations for a first AMI in young adults aged 18 to 44 years were identified from national inpatient sample from January 2005 through September 2015. Results: Chinese cohort included 2 866 young adults with a first AMI (male, n=2 739, female, n=127), the mean age was (39±5) years. Presentation of AMI was more frequently ST-segment elevation myocardial infarction (77.3%, 2 214/2 866). American cohort included 280 875 subjects (male, n=203 700, female, n=77 175), the mean age was 39±5 years. Presentation of AMI was more frequently non-ST-segment elevation myocardial infarction (53.6%, 150 549/280 875). In China, dyslipidemia 2 254 (78.6%), smoking 2 084(72.7%), and hypertension 1 170 (40.8%) were most prevalent, and 96.0% (2 752/2 866) of patients had at least 1 risk factor; in the United States, smoking 159 537(56.8%), dyslipidemia 145 212 (51.7%), and hypertension 139 876 (49.8%) were most prevalent, and 90.3% (253 630/280 875) of patients had at least 1 risk factor. Women had a prevalence of diabetes was higher in women, and prevalence of dyslipidemia and smoking was higher in men in China (all P<0.05);prevalence of obesity, diabetes and hypertension was higher in Women, and prevalence of dyslipidemia and smoking was higher in man in the United States (all P<0.001). Prevalence of dyslipidemia and smoking was higher Chinese men (79.3% vs. 54.6%, 75.5% vs. 58.1%,all P<0.001), and prevalence of obesity, diabetes and hypertension was lower (13.1% vs. 18.6%, 14.9% vs. 19.9%, 40.6% vs. 49.3%, all P<0.001)in Chinses cohort than those in the United States cohort. Prevalence of smoking and obesity was lower (12.6% vs. 53.4%, 10.2% vs. 26.9%, all P<0.001) and prevalence of dyslipidemia was higher (63.8% vs. 44.1% P<0.001) in Chinese women than those in the United States women. Patients presenting with non-ST-segment elevation myocardial infarction had a higher prevalence of obesity, diabetes and hypertension than patients presenting with ST-segment elevation myocardial infarction in China (20.1% vs.10.9%, 17.6% vs. 14.5%, 47.4% vs. 38.9%, all P<0.05). The prevalence of the three risk factors also was higher in patients presenting with non-ST-segment elevation myocardial infarction in the US (24.0% vs.17.0%, 25.0% vs. 20.0%, 54.6% vs. 44.2%, all P<0.001), prevalence of smoking and dyslipidemia was lower in these patients (53.5% vs. 60.5%,51.4% vs. 52.1%, all P<0.001). The prevalence of hypertension and obesity increased and the rate of smoking reduced in China from 2007 through 2017 (all trend P<0.001). The prevalence of all these five conventional risk factors increased temporally in the United States from 2005 to 2015 (all trend P<0.001). The prevalence of hypertension increased by 15.6% in China and 14.5% in the United States, respectively, accounting the largest increase. Conclusions: Smoking, dyslipidemia, and hypertension are most prevalent in China and United State. Significant sex and AMI subtype difference are observed for individual risk factors. The prevalence of hypertension and obesity increased significantly over time in China and all these five conventional risk factors increased significantly in the United States.

Bacillus velezensis WLYS23 strain possesses antagonistic activity against hybrid snakehead bacterial pathogens.

AIM: The aims of this study were to screen an antagonistic probiotic for the prevention and control of bacterial diseases in snakehead fish and to evaluate the antimicrobial activities, biosafety and biocontrol effect of the antagonistic strain. METHODS AND RESULTS: In this study, the WLYS23 strain exhibiting the most effective antagonistic properties against several fish pathogens was selected from the intestine of healthy snakehead fish. The strain was identified as Bacillus velezensis based on morphological, physiological and biochemical characteristics, and phylogenetic analysis. This strain showed amylase, cellulase, protease and lipase activities according to extracellular enzyme activity assays. Moreover, the cell-free extract of the WLYS23 strain exhibited strong antibacterial activity, with MICs of ≤64 µg ml-1 for most fish pathogens. Additionally, the cell-free extract was heat and pH stable, and resistant to protease, whereas their antimicrobial activities were significantly influenced by metal ions at high concentration. The WLYS23 strain and its cell-free extract were safe for snakehead and zebrafish. The disease resistance of snakehead fish was significantly increased following administration of the WLYS23 strain and its cell-free extract respectively. The complete genome of the WLYS23 strain was sequenced and found to harbour seven gene clusters encoding secondary metabolites with antimicrobial activity. CONCLUSION: The WLYS23 strain possesses great potential as a biocontrol agent, which can be commercially developed to improve disease control in freshwater aquaculture. SIGNIFICANCE AND IMPACT OF THE STUDY: Snakehead fish are important commercially farmed fish worldwide. However, the bacterial diseases caused by Aeromonas sp. and Nocardia seriolae in farmed snakehead fish lead to huge economic losses. Safe, economical and efficient probiotics are limited to prevent and control these diseases. Here, we provide a promising biocontrol agent with antagonistic activity against bacterial diseases of snakehead.

[Gender differences in risk factors and characteristics of young patients hospitalized for first acute coronary syndrome].

Objective: To analyze the characteristics of modifiable risk factors, coronary artery lesions, reperfusion and drug treatment in young women hospitalized for first acute coronary syndrome (ACS). Methods: Clinical data of young adults of 18 to 44 years hospitalized for first ACS in Beijing Anzhen Hospital between January 2007 and December 2017 were analyzed. A total of 7 106 young adults with ACS were enrolled, 6 593(92.8%) were male and 513(7.2%) were female. There were 2 254(31.7%) patients with ST-segment elevation myocardial infarction (STEMI), 704(9.9%) patients with non-ST-segment elevation myocardial infarction (NSTEMI) and 4 148(58.4%) patients with unstable angina (UA). Clinical characteristics, laboratory examinations, coronary angiography and medical treatment were recorded. The gender differences in the characteristics of modifiable risk factors and coronary artery stenosis in young patients with ACS were also analyzed. Results: Dyslipidemia (87.5%) followed by overweight/obese(83.8%) and smoking (68.5%) were most prevalent modifiable risk factors in men. Dyslipidemia (63.4%), overweight/obese (63.4%) and hypertension (45.5%) were most prevalent modifiable risk factors in women. The prevalence rates of at least 1 risk factor in men and women patients were 99.1% and 87.9%, respectively. Between 2007 and 2017, the rates of hypertension and overweight/obese in men increased, and the rates of dyslipidemia and smoking decreased (trend P<0.01). The changes of incidence of modifiable risk factors through the years in women were not statistically significant. The rate of men receiving coronary angiography was higher than that in women (96.6% vs 93.4%, P<0.01). Women were more likely to suffer left main coronary artery disease than men (7.3% vs 4.1%, P<0.01). Multi-vessel coronary artery disease occurred more in men (43.4%), and single vessel coronary artery disease occurred more in women (47.0%). Women presented with STEMI had a lower probability of receiving clopidogrel/ticagrelor (76.8% vs 86.1%, P<0.01) and angiotensin Ⅰ-converting enzyme inhibitor/angiotonin receptor blocker (ACEI/ARB) (46.5% vs 60.2%, P<0.01) than that in men. Women presented with UA had a lower probability of receiving aspirin (86.3% vs 89.9%), clopidogrel/ticagrelor (69.7% vs 75.6%), statin (78.8% vs 85.0%) and ACEI/ARB (32.7% vs 38.6%) than men (all P<0.01). Women had a higher probability of presenting Killip II-IV than men (38.6% vs 25.6%, P<0.05). Conclusion: The majority of young patients with ACS were men. The prevalence rates of modifiable risk factors are very high in both man and woman. Compared with man, women with AMI are more likely to develop acute heart failure with lower probability of reperfusion and drug treatment.

[Diagnostic value of non-invasive model for hepatic steatosis in patients infected with human immunodeficiency virus].

Objective: Hepatic steatosis has a high incidence in human immunodeficiency virus (HIV) infected people, and there is no effective non-invasive method to evaluate it. This study aims to evaluate the diagnostic value of non-invasive models for hepatic steatosis in this population. Methods: A single-center retrospective study was applied to evaluate: (1) the diagnostic value of controlled attenuation parameters (CAP) and hepatic steatosis index (HSI) in HIV-infected patients with hepatic steatosis; (2) the ability of the non-invasive model to distinguish hepatic steatosis caused by abnormal glucose and lipid metabolism and hepatic steatosis caused by hepatitis C virus infection; (3) the diagnostic value of the above models for hepatic steatosis in patients co-infected with HIV/hepatitis C virus. The diagnostic value of the model was analyzed and evaluated by diagnostic test and receiver operating characteristic curve. Results: (1) the diagnostic value of hepatic steatosis for HIV-infected patients: when CAP = 232 dB/m, the sensitivity and specificity were 89.2% and 78.1%, respectively; when HSI = 34, the sensitivity and specificity were 79.1% and 83.2%, respectively. (2) The ability to identify the causes of hepatic steatosis in HIV-infected patients: when CAP = 258dB/m, the sensitivity and specificity were 81.5% and 88.2%, respectively; when HSI = 37, the sensitivity and specificity were 70.7% and 92.4%, respectively. (3) The diagnostic value of hepatic steatosis in patients co-infected with HIV/hepatitis C virus: when CAP = 241 dB/m, the sensitivity and specificity were 80% and 71.4%, respectively; when HSI = 32, the sensitivity and specificity were 73% and 68.9%, respectively. Conclusion: CAP and HSI have superior diagnostic value for hepatic steatosis in patients infected with HIV.

[Finite element analysis of evaluating the change of the hip joint biomechanics following femoral neck shortening after cannulated screw fixation for osteoporotic femoral neck fracture].

Objective: To evaluate the stress status of femoral head and neck, screws and acetabulum caused by femoral neck shortening after internal fixation of femoral neck fracture with finite element method, and to analyze the stress of proximal femoral neck and acetabulum from the mechanical point of view. Methods: CT scan data of hip of a healthy adult female were collected. Three-dimensional reconstruction MICs and related module function simulation was used to establish the postoperative shortening model of femoral neck fracture with Pauwells angle>50°, which was treated with cannulated screws. The models were divided into four groups: normal femoral neck without shortening, shortening for 2.5 mm, shortening for 7.5 mm and shortening for 12.5 mm. The finite element analysis software MSC. Nastran2012 was used to do the mechanical analysis. The acetabulum surface, femoral head surface, proximal femur and the maximum stress, stress nephogram and other relevant data were collected. Results: The maximum tensile stress and the maximum stress at the fracture site of the femoral neck increased gradually with the increasing of shortening of femoral neck, however, the maximum compressive stress under the femoral neck and the medial cortex decreased gradually; the maximum stress on the surface of the femoral head was 14.9, 15.0, 16.3 and 16.3 MPa, respectively; the maximum stress on the surface of the acetabulum was 10.1, 10.1 and 10.5,11.7 MPa, respectively. Conclusion: The mechanical environment of the hip joint changes with femoral neck shortening. With the increasing of femoral neck shortening, the peak stress of the acetabulum increases continuously. When the femoral neck is shortened seriously, the load distribution is uneven and the complex mobility of hip joint is decreased. In addition, the change of shortening might play a role in the necrosis of femoral head.

AKT-GSK3ß Signaling Pathway Regulates Mitochondrial Dysfunction-Associated OPA1 Cleavage Contributing to Osteoblast Apoptosis: Preventative Effects of Hydroxytyrosol.

Oxidative stress (OS) induces osteoblast apoptosis, which plays a crucial role in the initiation and progression of osteoporosis. Although OS is closely associated with mitochondrial dysfunction, detailed mitochondrial mechanisms underlying OS-induced osteoblast apoptosis have not been thoroughly elucidated to date. In the present study, we found that mitochondrial abnormalities largely contributed to OS-induced osteoblast apoptosis, as evidenced by enhanced production of mitochondrial reactive oxygen species; considerable reduction in mitochondrial respiratory chain complex activity, mitochondrial membrane potential, and adenosine triphosphate production; abnormality in mitochondrial morphology; and alteration of mitochondrial dynamics. These mitochondrial abnormalities were primarily mediated by an imbalance in mitochondrial fusion and fission through a protein kinase B- (AKT-) glycogen synthase kinase 3ß- (GSK3ß-) optic atrophy 1- (OPA1-) dependent mechanism. Hydroxytyrosol (3,4-dihydroxyphenylethanol (HT)), an important compound in virgin olive oil, significantly prevented OS-induced osteoblast apoptosis. Specifically, HT inhibited OS-induced mitochondrial dysfunction by decreasing OPA1 cleavage and by increasing AKT and GSK3ß phosphorylation. Together, our results indicate that the AKT-GSK3ß signaling pathway regulates mitochondrial dysfunction-associated OPA1 cleavage, which may contribute to OS-induced osteoblast apoptosis. Moreover, our results suggest that HT could be an effective nutrient for preventing osteoporosis development.

Comparison of cardiac MRI with PET for assessment of myocardial viability in patients with coronary chronic total occlusion.

AIM: To compare cardiac magnetic resonance imaging (MRI) and positron-emission tomography (PET) assessment of myocardial viability in patients with coronary chronic total occlusion (CTO). MATERIALS AND METHODS: Eighty patients with coronary CTO underwent cardiac MRI and PET. Cardiac MRI images were analysed using a 17-segment model, and late gadolinium enhancement (LGE) and wall motion were scored. PET was used to classify myocardial viability via myocardial perfusion and 18F-fluorodeoxyglucose, digital superscript uptake. RESULTS: With PET as the reference standard, the sensitivity of cardiac MRI in detecting myocardial viability was 95.3%, specificity was 87.5%, positive predictive value was 96.8%, negative predictive value was 84.2%, and accuracy was 93.8% on a per patient basis. The receiver operator characteristic curve was used to analyse the performance of cardiac MRI in the detection of myocardial viability on a per-patient basis and the area under the curve was 0.910 (95% confidence interval 0.805 to 1). Cardiac MRI had the highest sensitivity and specificity for differentiating viable and non-viable myocardium as defined by PET when the cut-off value of LGE was 50%. The motion consistency and correlation of cardiac MRI and PET were analysed and kappa was 0.788 (r=0.825; p<0.001). CONCLUSION: Compared with PET, cardiac MRI assessment of myocardial viability in patients with coronary CTO has high sensitivity, specificity, and accuracy. Therefore, cardiac MRI can be used as an important method for evaluating myocardial viability in coronary CTO patients.

Development and validation of an individualized nomogram to identify occult peritoneal metastasis in patients with advanced gastric cancer.

BACKGROUND: Occult peritoneal metastasis (PM) in advanced gastric cancer (AGC) patients is highly possible to be missed on computed tomography (CT) images. Patients with occult PMs are subject to late detection or even improper surgical treatment. We therefore aimed to develop a radiomic nomogram to preoperatively identify occult PMs in AGC patients. PATIENTS AND METHODS: A total of 554 AGC patients from 4 centers were divided into 1 training, 1 internal validation, and 2 external validation cohorts. All patients' PM status was firstly diagnosed as negative by CT, but later confirmed by laparoscopy (PM-positive n = 122, PM-negative n = 432). Radiomic signatures reflecting phenotypes of the primary tumor (RS1) and peritoneum region (RS2) were built as predictors of PM from 266 quantitative image features. Individualized nomograms of PM status incorporating RS1, RS2, or clinical factors were developed and evaluated regarding prediction ability. RESULTS: RS1, RS2, and Lauren type were significant predictors of occult PM (all P < 0.05). A nomogram of these three factors demonstrated better diagnostic accuracy than the model with RS1, RS2, or clinical factors alone (all net reclassification improvement P < 0.05). The area under curve yielded was 0.958 [95% confidence interval (CI) 0.923-0.993], 0.941 (95% CI 0.904-0.977), 0.928 (95% CI 0.886-0.971), and 0.920 (95% CI 0.862-0.978) for the training, internal, and two external validation cohorts, respectively. Stratification analysis showed that this nomogram had potential generalization ability. CONCLUSION: CT phenotypes of both primary tumor and nearby peritoneum are significantly associated with occult PM status. A nomogram of these CT phenotypes and Lauren type has an excellent prediction ability of occult PM, and may have significant clinical implications on early detection of occult PM for AGC.

Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia, Oreochromis niloticus.

A disease with white spots in internal organs of Nile tilapia occurred in Zhanjiang, southern China. Multiple, white nodules, 0.8-2.2 mm in diameter, were scattered throughout the liver, spleen and kidney of diseased fish. Signs of nodules reproduced after artificial infection with the isolated strain. Isolated bacteria were Gram-negative, facultative anaerobic, motile, short rod-shaped, with a length of 1.2-2.2 µm. Morphological and biochemical tests, as well as phylogenetic analysis, all strongly indicated that the isolate from tilapia is identical to Aeromonas schubertii (A. schubertii) which temporary named LF1708 strain. Antibiotic sensitivity assays showed the LF1708 is sensitive to 24 of 27 tested antibiotics. Pathogenicity test revealed that the isolate at the dose of 3.75 × 106 CFU/g killed 100% of experimental tilapia within 2 days and the dose of 1 × 107 CFU/g killed 100% of experimental zebrafish within 1 day. Histopathology of diseased tilapia infected with A. schubertii showed numerous necrotic lesions widely distributed in spleen, liver and kidney, and infiltration with a large number of bacteria. To our knowledge, this was the first report that associated A. schubertii with mortality in tilapia.

Partial Recovery of Mitochondrial Function of Vitrified Porcine MII Stage Oocytes During Post-Thaw Incubation.

　　The survival of porcine oocytes is still very low after cryopreservation. OBJECTIVE: To investigate whether and when the mitochondrial function of vitrified porcine oocytes could be recovered post-thaw. MATERIALS AND METHODS: Mitochondrial potential, ROS level, ATP content, apoptotic rate, caspase activity, and parthenogenetics developmental ability of thawed porcine oocytes were measured after culture in vitro for 0, 1, 2 or 4 h. RESULTS: Mitochondrial potential after 2 h and 4 h post-thaw culture were 1.19 and 1.26, significantly lower than that of fresh oocytes but much higher than the groups cultured for 0 h and 1 h (P<0.05). Cryopreservation increased the ROS level in oocytes considerably, which decreased only after 2 to 4 h incubation following thaw. ATP content increased gradually over time and recovered to the level comparable to that of fresh oocytes after 4 h. Pan caspase levels increased after cryopreservation and reached the highest level at 1 h incubation. Thereafter it decreased to a low value, but still higher than fresh oocytes. Oocytes showing an early apoptotic event decreased upon 2 to 4 h incubation. The parthenogenetic cleavage and blastocyst rates were the highest (19.8% and 5.6%) after 2 h incubation. CONCLUSION: The recovery of mitochondrial function could complete after 2 to 4 h post-thaw incubation. Post-thaw incubation for 2 to 4 h reduced apoptotic events and improved parthenogenetic developmental ability of vitrified porcine MII stage oocytes.

A pleiotropic effect of the APOE gene: association of APOE polymorphisms with multibacillary leprosy in Han Chinese from Southwest China.

BACKGROUND: Patients with leprosy have a very low risk of Alzheimer disease (AD) and ß-amyloid (Aß) deposition is significantly lower in the brain tissue of elderly patients with leprosy compared with age-matched controls. Apolipoprotein E (ApoE) plays a critical role in lipid metabolic pathways and in the brain, facilitating the proteolytic clearance of Aß. We hypothesized that APOE confers risk of leprosy as lipid metabolism is involved in Mycobacterium leprae infection. OBJECTIVES: To investigate the potential genetic associations between APOE and leprosy in two independent Chinese case-control cohorts from the Yuxi and Wenshan prefectures, Yunnan Province of Southwest China. METHODS: Five APOE single-nucleotide polymorphisms (SNPs) were analysed in 1110 individuals (527 patients and 583 controls) from the Yuxi prefecture using a SNaPshot assay. Genetic variations in the entire APOE exons were screened in 1788 individuals (798 patients and 990 controls) from the Wenshan prefecture using next-generation sequencing technology. RESULTS: The AD-associated SNPs rs405509 and rs439401 increased the risk of leprosy per se and multibacillary leprosy (P < 0·005), but the APOE-ε4 allele did not. The SNPs rs405509 and rs439401 were cis expression quantitative trait loci (eQTL) for APOE expression in human skin. Differential APOE mRNA expression was observed in skin lesions of patients with type I reaction leprosy and those with multibacillary leprosy. APOE and related lipid genes are involved in an interaction network with leprosy susceptibility genes. CONCLUSIONS: The APOE gene is associated with leprosy, most likely by regulating lipid-metabolism-related genes.

[Interaction between family history of diabetes and hyperlipidemia on risk of diabetes in population with normotension in Harbin: a cross-sectional study].

Objective: To explore the interaction between family history of diabetes and hyperlipidemia on the risk of diabetes in population with normotension. Methods: A multistage stratified probability random sampling was conducted to select a representative sample of urban residents aged 20-74 years in Harbin. A total of 376 diabetes patients with normotension and 3 692 residents with normal blood pressure, normal fasting glucose, and normal 2 hours glucose from OGTT were surveyed. The interaction was evaluated by using crossover analysis and additive model. Results: Multivariate logistic regression analysis indicated that there was a possible additive interaction between family history of diabetes and hyperlipidemia on the risk of diabetes. The relative excess risk due to the interaction, the attributable proportion due to the interaction, and the synergy index were 1.97 (95%CI:-0.32-4.26), 0.30 (95%CI: 0.03-0.57), and 1.54 (95%CI: 0.96-2.47), respectively. There were significant combination effects between family history of diabetes and high both total cholesterol and triglyceride, isolated high total cholesterol, and isolated high triglyceride levels; the ORs were 10.55 (95%CI: 5.62-19.80), 7.81 (95%CI: 3.65-16.71) and 5.13 (95%CI: 3.22-8.16), respectively. Conclusion: There might be synergistic effect between family history of diabetes and hyperlipidemia on the risk of diabetes in population with normotension.

Direct sequencing of mutations in the copper-transporting P-type adenosine triphosphate (ATP7B) gene for diagnosis and pathogenesis of Wilson's disease.

Copper-transporting P-type adenosine triphosphatase (ATP7B) has been identified as the pathogenic gene in hepatolenticular degeneration, or Wilson's disease (WD). The aim of this study was to explore the correlation between genetic mutations and the clinical profile of WD, and to discuss the value of mutation examination in its diagnosis for providing a scientific basis for the development of a method to examine genetic mutations. Sixty-eight Chinese Han patients with WD and 20 controls were included in this study. The ATP7B gene in DNA extracted from patient blood samples was amplified by PCR and sequenced. These sequences were compared against corresponding gene sequences obtained from healthy controls to statistically analyze the genetic mutations. Five of the nineteen mutations in ATP7B were newly detected mutations; moreover, 8 of these mutations were polymorphic (2 were newly identified). The Arg778Leu and Pro992Leu mutations in exons 8 and 13 were detected at the highest mutation frequencies of 25.74 and 16.91%, respectively. The frequencies of all other mutations were below 5%. However, the clinical manifestations of WD did not differ significantly in patients with the Arg778Leu and Pro992Leu mutations. Therefore, these mutations were considered as hotspot mutations in Chinese WD patients. However, we observed no significant correlation between these genetic types and the clinical symptoms of WD. The correlation between the mutation genotype and disease phenotype remains to be elucidated. In conclusion, the highly sensitive and specific direct DNA sequencing method can be used to screen for the causative genes of WD.

Both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified mii stage oocytes.

BACKGROUND: Oocytes vitrification is widely used for cryopreservation of female genetic resources. OBJECTIVE: In order to illuminate the apoptotic pathways of porcine MII stage oocytes after vitrification. MATERIALS AND METHODS: This study used in situ fluorescence staining and RT-PCR to detect the expression levels of some key molecules from death receptor and mitochondria mediated apoptotic pathways. RESULTS: (1) Early stage apoptosis were detected in both PI staining survival oocytes and PI staining dead oocytes. (2) The fluorescence intensity of caspase 8, caspase 9, caspase 3 and pan caspase from vitrified oocytes were 32.03, 16.56, 16.70 and 8.43 respectively, which were much higher than those from fresh oocytes (4.02, 4.83, 4.23 and 3.08, P < 0.05). (3) Not only the genes from death receptor mediated apoptotic pathway, but also from mitochondrial mediated apoptotic pathway were changed greatly. CONCLUSION: The death of porcine vitrified oocytes could be induced by apoptosis, both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified MII stage oocytes.

TGF-ß1 and Serpine 1 expression changes in traumatic deep vein thrombosis.

The objective of this study was to investigate the expression changes of transforming growth factor ß1 (TGF-ß1) and Serpine 1 in rats with traumatic deep vein thrombosis (DVT). In total, 60 male Sprague Dawley rats were divided into model (N = 50) and control groups (Group A, N = 10). From the model group, 10 rats were randomly selected after modeling as the pre-thrombosis group (Group B, N = 10), and the remaining 40 rats in the model group were divided into the thrombosis (Group C) and no thrombosis groups (Group D) depending on whether DVT was apparent at 25 h after modeling. All rats were dissected and the total RNAs of the femoral veins were extracted. TGF-ß1 and Serpine 1 expression was detected by microarray and polymerase chain reaction (PCR) analyses, and the related signal pathways were analyzed using bioinformatic analysis. Of the 40 rats, DVT was evident in 23, yielding an incidence rate of 57.50%. TGF-ß1 and Serpine 1 expression increased significantly at 2.5 h after modeling, while DVT began to form at 25 h after modeling. Both PCR and microarray analysis showed that TGF-ß1 and Serpine 1 expression levels were significantly higher in the thrombosis group than in the other groups (P < 0.05). Bioinformatic analysis indicated that TGF-ß1 was an upstream regulatory gene of Serpine 1 and could induce Serpine 1 overexpression. Together, these results suggested that TGF-ß1 and Serpine 1 overexpression might play an important role in DVT formation and have predictive values.

Detection of targets and their mechanisms for early diagnosis of traumatic deep vein thrombosis.

The purpose of this investigation was to identify targets for the early diagnosis and predictors of deep venous thrombosis (DVT) and the role of these targets in the formation of venous thrombosis. A model of DVT was constructed in rats. Thromboses and venous walls were sampled for reverse transcription polymerase chain reaction study, and blood was sampled for enzyme-linked immunosorbent assay studies. Vein endothelial cells were cultured to observe the effects of interleukin (IL)-17 on the expression of tissue plasminogen activator (t-PA)/plasminogen activator inhibitor type 1 (PAI-1). IL-17 monoclonal antibody was used to study its effect on preventing the formation of DVT. One-hundred and twenty hours after the animal model was constructed, significant DVT started to form. Polymerase chain reaction tests showed that immediately after the model was created, the expression of IL-17 increased greatly, whereas the balance between t-PA and PAI-1 was disrupted just before DVT formed. The increase of serum IL-17 was positively related with the formation of DVT. Thus, the application of IL-17 monoclonal antibody could reduce the formation of DVT in rats. IL-17 might be a target for the early diagnosis of DVT and should be further studied to assess its clinical value.

Association of the LRRK2 genetic polymorphisms with leprosy in Han Chinese from Southwest China.

Leprosy is a chronic infectious and neurological disease that is caused by infection of Mycobacterium leprae (M. leprae). A recent genome-wide association study indicated a suggestive association of LRRK2 genetic variant rs1873613 with leprosy in Chinese population. To validate this association and further identify potential causal variants of LRRK2 with leprosy, we genotyped 13 LRRK2 variants in 548 leprosy patients and 1078 healthy individuals from Yunnan Province and (re-)analyzed 3225 Han Chinese across China. Variants rs1427267, rs3761863, rs1873613, rs732374 and rs7298930 were significantly associated with leprosy per se and/or paucibacillary leprosy (PB). Haplotype A-G-A-C-A was significantly associated with leprosy per se (P=0.018) and PB (P=0.020). Overexpression of the protective allele (Thr2397) of rs3761863 in HEK293 cells led to a significantly increased nuclear factor of activated T-cells' activity compared with allele Met2397 after lipopolysaccharides stimulation. Allele Thr2397 could attenuate 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine-induced autophagic activity in U251 cells. These data suggest that the protective effect of LRRK2 variant p.M2397T on leprosy might be mediated by increasing immune response and decreasing neurotoxicity after M. leprae loading. Our findings confirm that LRRK2 is a susceptible gene to leprosy in Han Chinese population.