Objective: To investigate the effects of polydatin on a high-fat diet-induced non-alcoholic fatty liver disease(NAFLD) mouse model and hepatoma G2(HepG2) cell model, and to reveal its potential molecular mechanisms. Methods: Thirty 6-week-old male SPF C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet group. After the NAFLD mouse model was established in the high-fat diet group, they were further divided into a model group and a polydatin treatment group. The polydatin treatment group was administered polydatin by gavage at a dose of 250 mg/(kg·d) for 10 weeks, during which body weight was monitored and oral glucose and insulin tolerance tests were performed. At the end of the experiment, a series of tests to evaluate the effects of polydatin on mouse liver weight, blood lipids, liver lipid accumulation, and liver injury markers were performed. The expression of G0/G1 switch gene 2(G0S2) and adipose triglyceride lipase(ATGL) was measured by qRT-PCR and Western blot, and gene expression was further verified using immunohistochemical staining. The effects of polydatin on HepG2 cell activity was assessed by CCK-8 assay, lipid accumulation was observed by oil red O staining, and the expression of G0S2 and ATGL was detected by qRT-PCR and Western blot. Results: Polydatin significantly reduced the body weight, liver weight, and serum and liver tissue levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), triglyceride(TG), and total cholesterol (TC) in mice (P < 0. 05) , al⁃leviated pathological liver damage , decreased G0S2 expression (P < 0. 05) , and increased ATGL expression (P <0. 05) . At the cellular level , polydatin reduced lipid droplet accumulation , improved lipid metabolism , decreased G0S2 expression ( P < 0. 05 ) , and increased ATGL expression ( P < 0. 05 ) . Even in cells with knockdown of G0S2 , polydatin still promoted fat decomposition (P < 0. 01) . Conclusion Polydatin promotes hepatic fat break⁃down by regulating the expression of G0S2 and ATGL , helping to alleviate metabolic disorders and liver damage in the NAFLD mouse model caused by a high⁃fat diet , offering a new strategy for treating NAFLD.

Modern TCM understands the pathogenesis of blood glucose fluctuation mainly from the viscera (spleen, kidney, liver, bile, small intestine), middle energizer, "yin fire theory", "Xuanfu depression" and "qi transformation". In clinic, blood glucose fluctuation is mainly divided into four syndrome types: qi-yin deficiency syndrome, yin-yang deficiency syndrome, exuberance of fire and heat syndrome, and heat stagnation in liver-stomach syndrome. TCM compounds can improve blood glucose fluctuation by improving insulin resistance, regulating intestinal microflora, promoting the secretion of glucagon-like peptide-1, and fighting oxidative stress.

Mutants of proteins are the basis for studying their structure and function, this work aimed to establish an efficient and rapid method for constructing multi-site mutants. When four or more adjacent amino acid residues need to be mutated, firstly, two long and two short primers (long primers Ⅰ/Ⅰ, short primersⅡ/Ⅱ) were designed: the long primers contain mutated sites, and the number of mutant bases is ≤20 bp, the short primers do not contain mutated sites; GC contents of the long and short primers are ≤80%, and the difference of annealing temperature is ≤40 °C. Then two sets of reverse PCR amplifications were performed using primer pairs (Ⅰ/Ⅱand Ⅰ/Ⅱ) and templates, respectively. After amplification, each system can obtain non-methylated linear plasmids which contain mutated sites, and the breakpoints of the two sets of linear plasmids amplified by primers Ⅰ/Ⅱ and Ⅲ/Ⅳ were distributed on both sides of the mutated sites. Followed by digested by DpnⅠ to remove the methylated templates, the recovered PCR products, which were mixed in an equimolar ratio, were performed another round of denaturation and annealing: the two sets of linear plasmids were denatured at 95 °C and then annealed with each other's single-stranded DNA as templates to form open-loop plasmids, and then the transformants containing the mutations will be obtained after transformed the open-loop plasmids into Escherichia coli competent cells. Results showed that, this method can mutate 4 to 11 consecutive amino acid residues (8-20 bp) simultaneously, which will greatly simplify the construction of multi-site mutants, Thereby improve the efficiency of protein structure and function research further.
DNA Primers ; genetics ; Escherichia coli ; Mutagenesis, Site-Directed ; methods ; Plasmids ; genetics ; Polymerase Chain Reaction

Objective To establish an athymic (nu/nu)BALB/c mouse model with chronic subcutaneous infection due to Phialophora verrucosa (P.verrucosa),and to explore the role of T lymphocytes in defensing against invasive infection due to P.verrucosa.Methods Six immunocompetent BALB/c mice and 6 nu/nu BALB/c mice were subcutaneously inoculated with 100 μl of P.verrucosa hyphae suspensions at a concentration of 5.0 × 108 colony-forming unit (CFU)/ml into one footpad,while another 6 immunocompetent BALB/c mice and 6 nu/nu BALB/c mice were subcutaneously inoculated with 100 μl of 5.0 × 108 CFU/ml P.verrucosa conidium suspensions into one footpad.The degree of footpad swelling was measured with a vernier caliper every 3 days.Histopathological characteristics of infected footpads were further analyzed.Biopsy tissues were also subjected to fungal culture to analyze the growth of P.verrucosa in infection foci in mice.Results After the treatment with hyphae or conidium suspensions,the BALB/c mice experienced transient footpad swelling within 12 days,and basically recovered after 50 days.However,the nu/nu BALB/c mice experienced persistent footpad swelling with recurrent ulceration and crusting.As pathological examination revealed,all the inoculation loci in BALB/c mice experienced local infection,and the morphology of P.verrucosa in the infected foci was not changed over time.However,invasive infections due to P.verrucosa hyphae alone or a mixture of P.verrucosa hyphae and sclerotic cells were observed in all the inoculation loci in nu/nu BALB/c mice.The fungal culture showed that P.verrucosa could not grow in the footpads of BALB/c mice after 21 days,while P.verrucosa could persistently grow in the footpads of nu/nu BALB/c mice.Conclusion An experimental model with subcutaneous phaeohyphomycosis due to P.verrucosa has been successfully established by using nu/nu BALB/c mice,and the nu/nu BALB/c mice are more susceptible to P.verrucosa infection compared with the immunocompetent BALB/c mice.

Objective To compare the application of Goldman , CARPREG and ZAHARA in predicting cardiac outcomes in pregnant women with congenital heart disease. Methods A retrospective analysis of 144 pregnant women with CHD , enrolled in The Third Affiliated Hospital of Guangzhou Medical University between Jan 1992 and June 2015 was conducted. The mortality and the incidence of cardiac events were counted by three risk estimation methods. The sensitivity and calibration of three systems were compared by using Hosmer-Lemeshow goodness of-fit analysis and Aera Under Curve (AUC). Results In terms of mortality rates, the sensitivity of CARPREG was higher than that of Goldman (AUC:0.874 and 0.889 respectively;P = 0.568 >0.05);. In terms of the incidence of cardiac events, the sensitivity of ZAHARA was higher than that of Goldman (AUC:0.912 and 0.773 respectively;P = 0.063 > 0.05); Conclusions All 3 risk estimation methods have good performance in sensitivity of predicting the incidence of cardiac events and mortality in pregnant women with CHD. The ZAHARA has a better calibration in predicting the incidence of cardiac events , and we recommend ZAHARA as the first choice of evaluation system for this kind of patients.

OBJECTIVE:To provide reference for safe application of antibiotic drugs in single or inter-medical institution ser-vices. METHODS:An antibiotic drug safety evaluation index system was established by reference to literatures. Medical records tracing method was adopted to obtain the medical records of children with respiratory infectious diseases under the medical consor-tium model. The medical records were matched and evaluated by experts to analysis the safety of antibiotic drug use in single medi-cal institution and the connection between two medical institutions with respect to the safety of antibiotic drug use. RESULTS:248 effective medical records had been obtained. Under the medical consortium model,the rate of the combination of antibiotic drugs in large hospitals was 86.29%,higher than the community hospitals of 77.02%(χ2=5.49,P<0.05). The error rate of antibiotic drug use by children in large hospitals was 14.52%,lower than the community hospitals of 25.81%(χ2=9.733,P<0.05). There were many contradictions in antibiotic drug use between the medical institution where children received treatment initially and the medi-cal institution which the children were referred to and hospitalized in. There were totally 128 cases of unsafe antibiotic drug use, with the overall incidence of 51.61%. CONCLUSIONS:Under the medical consortium model,the safety of antibiotic drug use by the hospitalized children in single medical institution is worrying,and the connection between two medical institutions with respect to the safety of antibiotic drug use by children who are referred and hospitalized is less reliable.

BACKGROUND: It has been explained in many big sample experiments that community comprehensive prevention from the risks of cardiocerebrovascular disease can reduce its morbidity and mortality. But, it is required more samples to verify the evaluations of the process and effect of intervention.OBJECTIVE: To understand the effect of community comprehensive prevention on cardiocerebrovascular disease and hygienic costs demanded and summarizes the suitable patterns of comprehensive prevention on cardiocerebrovascular disease in countryside.DESIGN: Healthy people in community were taken as the objects and community intervention was designed.SETTING: Department of Community Hygienic Service of People's Hospital in Guzheng Town, Zhongshan City, Guangdong Province and Neurological Institute of Guangzhou Medical College.PARTICIPANTS: Totally 32 000 inhabitants were collected from 8 villages in Guzheng Town, Zhongshan City, Guangdong Province from March 1992 to March 2002, aged varied from 20 to 74 years, of which, 14 600inhabitants were male and 17 400 inhabitants were female.METHODS: The intervention of community-based prevention was applied on cardiocerebrovascular disease, including managementof hypertension,smoking control, rational nutrient, nationwide body building and diabetic management.MAIN OUTCOME MEASURES: [1] Awareness, attitude and behavior related to cardiocerebrovascular disease. [2] Management rate and control rate of hypertension. [3] Morbidity and mortality of cerebral apoplexy.RESULTS: [1] By 10-year intervention, awareness rate of hypertension in whole group was 70.29%. [2] Management rate and control rate of hypertension were increased yearly, in which, the management rate was increased from 25.66% in 1997 to 80.50% in 2002, the total control rate was increased from 4.43% in 1997 to 45.80% in 2002, the control rate of blood pressure ＜160/95 mm HG (1 mm Hg=0.133 kPa) was 58.80%and that ＜140/90 mm Hg was 32.80%. [3] The systolic and diastolic pressures in intervention community were decreased averagely, in which the D-values are 7.23 mm Hg and 3.92 mm Hg respectively. [4] The awareness, attitude and behavior related to cardiocerebrovascular disease were enhanced remarkably. [5] Morbidity and mortality of cerebral apoplexy were in tendency of decreasing yearly, from 146.90/100 000 and 108.63/100 000 in 1997 to 105.83/100 000 and 69.90/100 000 in 2002 respectively, by 41.07/100 000 and 38.73/100 000 respectively in 6 years and the age of incidence was postponed. [6] It was indicated in economic analysis that the average ratio of profit to cost between 1997 and 2001 was 2.32.CONCLUSION: Community-based comprehensive prevention on cardiocerebrovascular disease improves the awareness, attitude and behavior in community group and reduces morbidity and mortality of cerebral apoplexy.