Objective To investigate the effect of Sirt1 gene knockout on chronic kidney disease induced by 5/6 nephrectomy in mice and vascular endothelial growth factor (VEGF)/fetal liver kinase-1 (Flk-1) signaling pathway.Methods Twenty four male Sirt1 +/+ and Sirt1 +/-mice wererandomly divided into four groups:Sirt1+/+ mice with sham-operation (WT-Sham,n=6),Sirt1+/-mice with sham-operation (KO-Sham,n=6),Sirt1 +/+ mice with 5/6 nephrectomy (WT-Nx,n=6) and Sirt1 +/-mice with 5/6 nephrectomy (KO-Nx,n=6).Proteinuria was determined by urine collection from 8:00 to 8:00 the next day at 20 weeks.Serum creatinine (Scr),urea nitrogen (BUN) and the renal pathological changes were measured after 20 weeks.Expressions of Sirt1,collagen Ⅰ and transforming growth factor β(TGF-β) were used to analyze the changes of renal fibrosis by immunohistochemistry staining.Real-time PCR and Western blotting were used to measure the mRNA and protein expressions of Sirt1,fibronectin,collagen Ⅰ,VEGF and Flk-1 in kidney.Results Sirt1 expressed in glomernlar endothelial cells,podocytes,mesangial cells and renal tubular epithelial cells in Sirt1 +/+ mice,while Sirt1 expression intensity was significantly reduced in Sirt1 +/-mice.Compared with the WT-Sham group,WT-Nx group had increased proteinuria,BUN,Scr,glomernlar sclerosis index and tubulointerstitial fibrosis index at 12 weeks after operation (all P ＜ 0.01),and KO-Nx group had exacerbated the above up-regulations (all P ＜ 0.01).Compared with those in WT-Sham group,the expressions of fibronectin,collagen Ⅰ and TGF-β were up-regulated in WT-Nx group (all P ＜ 0.01),and were significantly augmented in KO-Nx group (all P ＜ 0.01).Compared with those in WT-Sham group,renal mRNA and protein expressions of VEGF and Flk-1 were decreased in WT-Nx group,and KO-Nx group aggravated their down-regulation (all P ＜ 0.01).Conclusions Sirt1 gene knockout can increase proteinuria and Scr,and aggravate renal pathology and renal fibrosis in 5/6 nephrectomized mice,which is associated with the inhibition of VEGF/Flk-1 signaling pathway.It is suggested that Sirt1 may be a potential therapeutic target of chronic kidney disease.

Objective: To study the hepatitis E virus(HEV) infection in pregnant women and the healthy blood donors in Jinan. Methods: A total of 651 blood samples from pregnant women who came for screening of hemolytic disease of newborn and 600 blood samples from blood donors were collected during June 2015 to October 2016. All the blood samples were tested for anti-HEV IgG and anti-HEV IgM antibodies by using enzyme-linked immunosorbent assay (ELISA). Results: Anti-HEV IgG was positive in 2.61% (17/651) of the blood samples from the pregnant women, and none of the samples were positive for anti-HEV IgM. The serum positive rates for anti-HEV IgG and IgM in blood donors were 16.33% (98/600) and 0.83% (5/600) respectively. The seroprevalence of the two groups was significantly different(χ²=70.43, P<0.0001). Conclusions The HEV infection rate in pregnant women was much lower compared with that in blood donors in Jinan; however, in view of the high mortality of HEV infection in pregnant women, it still cannot be neglected. The infection of HEV in blood donors of Jinan was confirmed, suggesting the potential risk of transfusion-transmitted HEV infection.

Objective To establish cell strain expressing the genes of HIV vpr and mutant HIV vpr-FS, and to explore cell apoptosis ability by HIV Vpr and Vpr-FS. Methods The recombinant plasmids were constructed by cloning HIV vpr and HIV vpr-FS genes into the eukaryotic expression vector pcDNA3.1respectively. To determine the primary structures of HIV vpr and HIV vpr-FS, plasmids were cleaved by restriction enzymes. After the plasmids were transfected into HeLa cells by liposome, the HeLa cells were selected with G418 selective medium, mRNA expression of HIV vpr or HIV vpr-FS of transfected cells was detected by RT-PCR, and Vpr and Vpr-FS protein expression were detected by Western blot assay respectively. The DNA content and the percentage of apoptosis in HeLa HIV vpr cell, HeLa HIV vpr-FS cell and HeLa pcDNA3.1 cell were monitored by flow cytometry and the DNA fragmentation was analyzed by agarose gel electrophoresis. Results BamH Ⅰ and Hind Ⅲ cleavaged products of pcDNA3.1-vpr and pcDNA3.1-vpr-Fincluded 342 bp length fragments suggesting that the length of DNA sequence containing HIV vpr (HIV vpr-FS) within pcDNA3.1 was the same as theoretical length. The HeLa cells transfected by pcDNA3.1-vpr or pcDNA3, l-vpr-FS and selected with G418 could express HIV vpr or HIV vpr-FS by RT-PCR, and express HIV Vpr or HIV Vpr-FS protein by Western blot. The results of flow cytometry and DNA fragmentation showed that there was significant different in the number of apoptotic cells between HeLa HIV vpr cell and HeLa HIV vpr-FS cell, but the difference between HeLa HIV vpr-FS cell and control group was not obvious. Conclusion Recombinant plasmids pcDNA3.1-vpr and pcDNA3. 1-vpr-FS were constructed successfully, and the cell strain expressing HIV Vpr and HIV Vpr-FS proteins was established. The HIV Vpr could induce host cell apoptosis, while the mutant of Vpr did not or weakened this ability. This study provides foundation for further study on HIV vpr gene.

Objective: To investigate milk consumption and associated factors in the national pilot counties of Nutrition Improvement Programme for Rural Compulsory Education Students (NIPRCES) in 2016, and to provide basic data for milk consumption improvement. Methods: Two primary and middle schools in each county were selected randomly from 50 key monitoring counties in 22 provinces in NIPRCES by three food supply modes (school-based, company-based, and others). Then 1 or 2 classes in each grade were selected randomly from grade 3 or above and junior schools. A total of 25 948 students were collected, milk consumption and related factors were investigated through student questionnaires. Results: Among the students surveyed, the proportion of students drank 1 to 3 packs of milk every week was highest (45.4%), only 29.1% students drank 4 or more packets of milk per week. The proportion of children in grades 3 and 4 who drank 4 or more packets of milk per week (35.7%) was higher than that of children in grades 5 and 6 (31.8%) and junior school (22.5%)(P<0.01). The proportion of boys who drank 4 or more packs of milk per week (30.7%) was higher than girls (27.5%) (P<0.01). Multivariate Logistic regression analysis showed that boys, primary school students in grades 3 to 6, non-resident students, students with both parents or one parent at home, students who had more meals per day, students who had breakfast every day, students who took snacks or drank beverages once or more per day, students who drank 4 or more cups of boiled water were more likely to drink 4 or more packs of milk each week. Conclusion Students in poor rural areas in China lack access to milk. It is recommended that targeted measures should be taken to encourage and promote rural students to drink enough milk.

Objective To investigate the curative effect of ultra early enteral nutrition (EN) supplemented with probiotics in patients with severe acute pancreatitis (SAP).Methods Seventy-five SAP cases admitted in Second People's Hospital of Nantong from December 2014 to June 2018 were enrolled and assigned into 24 h group (who received EN & probiotics at 24 h,n =25),72 h group (who received EN & probiotics at 72 h,n =25) and the control group (who received EN & probiotics on day 7,n =25).After admission,the levels of PCT,hs-CRP and endotoxin on day 1,4 and 8 were detected.APACHE Ⅱ score on day 3 after admission,complication rate,infection rate,hospitalization duration and mortality were recorded.Results On day 1 after admission,serum PCT,hs-CRP and endotoxin concentrations among three groups were of no significant differences.On day 4 after admission,the serum concentrations of PCT and edotoxin in 24 h group were significantly lower than those in 72 h group and the control group [(3.12 ± 1.45) μg/L vs (5.26±1.52),(6.07 ± l.59) μg/L;(0.24 ± 0.02) EU/ml vs (0.35 ±0.03),(0.46±0.04) EU/ml].The serum hs-CRP in 24 h group was significantly lower than that of the control group [(20.71 ±4.89) μg/L vs (28.37 ± 4.64) μg/L],and the difference was statistically significant (P ＜ O.05).The serum concentrations of hs-CRP in 24 h group and 72 h group was of no significant difference.On day 7 after admission,the serum concentrations of PCT,hs-CRP and endotoxin in 24 h group were significantly lower than those in 72 h group and the control group (P ＜0.05);the serum concentrations of PCT,hs-CRP and endotoxin in 72 h group were significantly lower than those of the control group,and all the differences were statistically significant (all P ＜ 0.05).The score of APACHE Ⅱ (day 3 after admission) [(15.1 ± 1.8)],complication rate (52％),infection rate (8％),length of stay [(19.7 ± 5.3) d] in 24 h group were all significantly lower than those of 72h group [(17.0 ± 2.0),72％,32％,(25.4 ± 6.8) d] and control group [(18.3 ±2.6),84％,44％,(38.7 ± 12.6) d],and all the differences were statistically significant (all P ＜0.05).Conclusions Ultra EN supplemented with probiotics in treating the patients with SAP could significantly reduce the level of serum PCT,hs-CRP and endotoxin,decrease complication rate and mortality,and shorten the time of hospitalization.

AIM:We investigated how AT 1-R stimulated by mechanical stresses induces cardiac fibrosis .METHODS:We produced in vivo cardiac pressure overload model in angiotensinogen knockout ( ATG-/-) mice and in vitro mechanically-stretched cell model in cultured neonatal cardiac cells of ATG-/-mice both lack the participation of Ang II .RESULTS: Pressure overload for 4 weeks in ATG-/-mice induced myocardial hypertrophy accompanied by the significant interstitial fibrosis , however , the TGF-β, a key regulatory factor of fibrosis, was not significantly increased in these ATG-/-mice.Meanwhile, the inhibitor for AT1-R significantly inhibited mechani-cal stress-induced cardiac fibrosis in these ATG-/-models whereas inhibition of TGF-βdid not.CONCLUSION:The results showed that mechanical stress-induced fibrotic responses through AT 1-R required the phosphorylation of Smad 2 but not the involvement of TGF-β.

Objective: To explore the prevalence of anemia and related factors among left-behind children in poverty-stricken rural areas in China, to provide basic information for anemia prevention and treatment for children. Methods: Data from the National Nutrition Improvement Program for Rural Compulsory Education Students in 2016 were used to express the prevalence of anemia among left-behind children. By using the multistage stratified random cluster sampling method, left-behind children of the 6-17-years-old in poverty-stricken rural areas were recruited in this study. Blood hemoglobin concentration of these students was determined by cyanmethemoglobin method. Anemia was judged by the WHO recommended standard and combined with the sea level elevation correction standard. Frequencies of food intake were collected through questionnaires. Data was then available for analysis including the level of hemoglobin and anemia rates. Relationship between anemia and food intakes as well as other influential factors was also analyzed. Results: The overall anemia prevalence (AP) was 11.6% among the participants. Rates of AP among the 6-, 7-, 15- and 16-17-years-old were 21.1%, 18.0%, 20.6% and 17.3%, respectively. Rates of AP among left-behind girls of 10-11-years-old and the 13-15-years-old were significantly higher than those in boys at the same age (P<0.01) group. Factors including: being girls (OR=1.34, 95%CI: 1.23-1.45), at grade 7 to 9 (OR=1.47, 95%CI:1.30-1.67) and mother being migrant worker (OR=1.32, 95%CI:1.16- 1.50) had higher proportions of getting anemia. Factors as living in the western area (OR=0.85, 95%CI: 0.78-0.93), taking breakfast every day (OR=0.70, 95%CI: 0.64-0.76), having meat more than twice per week (OR=0.84, 95%CI: 0.75-0.93) and having two kinds of vegetables per day (OR=0.85, 95%CI: 0.75-0.96) appeared protective. Conclusions Prevalence of anemia in left-behind children from the poverty-stricken rural areas was high, including junior middle school students from grade 7 to 9 at the early stage of youth development, girls and children with mothers as migrant workers. Anemia should be reduced by promoting the protective factors as having breakfast, increasing intakes of meat and fresh vegetables.

Objective: To analyze the height changes among children who had received subsidies of the National Nutrition Improvement Program for Rural Compulsory Education Students (NNIPRCES) by comparing data from the year of 2012 and 2017. Methods: Among 699 counties where NNIPRCES was implemented, at least 10% of elementary schools and primary middle schools with different food supply modes (school canteen meals, company meals, and mix meals) in the county were selected randomly. Forty students from one or two classes from each grade (grade 1 to grade 9) were randomly selected, to ensure equal proportion of boys and girls in each grade were surveyed, data on height from 1 655 793 students measured in 2012 were compared with 1 419 281 students measured in 2017. Height was measured by metal height pole within 0.1 cm accuracy. Results: From 2012 to 2017, the average increase of height in boys and girls aged 6-15 years was 1.9 and 2.0 cm, respectively. Among them, maximum increment was found in boys at the age of 13 years and in girls at the age of 12 years, respectively, which was 3.0 and 2.8 cm. The average increase in boys from the central and western region was 2.1 and 2.2 cm, respectively, while increase in girls from the central and western region was both 2.2 cm. Compared with the results from 2012, height of boys with the 5th, 50th and 95th height percentiles in 2012 increased by 1.7, 2.0 and 2.1 cm, respectively in 2017, while the increase in girls was 2.1, 2.0 and 2.2 cm, respectively. The average increment of western students with the 5th and 95th height percentiles was higher than those of students from the central region. Conclusion The average height of students has increased after the implementation of NNIPRCES. However, the development was unbalanced between the central and western region, which requires more targeted intervening strategies to improve the nutritional status of students.

Objective: To analyze the usage of catering software and the creation of recipes by schools, as part of the Nutrition Improvement Program for Rural Compulsory Education Students (NIPRCES) 2013-2017. Methods: In each county of the 699 trial counties across 22 provinces in central and western area of China under the NNIPRCES, no less than 10% of primary and junior middle schools were randomly selected according to their catering mode (i.e., school feeding, food packages, and family feeding). School questionnaires were used to collect information concerning the schools and the creation of recipes. Results: From 2013 to 2017, the utilization rate of catering software was 11.7%, 8.0%, 17.8%, 16.9%, and 14.0%, respectively with significant differences(χ 2=345.09，P<0.01). The creators of school-meal recipes included schools, the Education Bureau, hospitals and colleges, the centers of disease prevention and control. Differences were observed in the proportion of recipe creators across school type, area, and catering mode, while annual trends also varied (P<0.01), as indicated by the following proportions: 74.9%, 20.0%, 3.7%, 1.3%, respectively. Food safety was identified as the main factor that needs to be considered when creating school recipes, the proportions of which were 58.0%, 78.4%, 70.6%, and 87.4% from 2014 to 2017. Conclusion From 2013 to 2017, the utilization rate and the frequency of catering software were both relatively low, and recipe creation was in need of professional guidance. It is necessary to strengthen the popularization of catering software and improve the nutritional knowledge and skills of canteen workers to ensure the quality of school meals.

Objective:To explore the effects of competency-based diversified training on the bedside nursing comprehensive ability aand transition shock of newly recruited nurses.Methods:A total of 62 newly recruited nurses from the Second Affiliated Hospital of Zhengzhou University in June 2018 were selected as the control group, and 70 newly recruited nurses from June 2019 were selected as the experimental group by cluster sampling. The control group received routine pre-job training, and the experimental group received diversified training based on the control group. After one year of training, the differences in the scores of bedside nursing comprehensive ability, post competency and transition shock between the two groups of nurses were analyzed.Results:After 1 year of training, the total scores and the scores of nursing problems, nursing evaluation, nursing plan, nursing theory, nursing operation and bedside nursing comprehensive ability in the experimental group were 12.20±1.76, 24.29±2.38, 13.54±1.88, 9.77±1.35, 15.14±2.30 and 79.24±6.59, which were all higher than those of the control group (10.26±1.55,22.31±1.84,10.45±1.58, 8.56±1.80, 13.27±2.38 and 69.82±4.09), the differences were statistically significant ( t value was 4.582-13.679, P<0.05). The total scores and the scores of each dimension of the Post-competency the experimental group were 31.46 ±4.32, 51.03± 4.64, 20.27± 3.78, 17.86±3.99, 58.43± 4.44, 179.37±9.97, which were all higher than those of the control group 24.76±4.12, 40.56±5.55, 16.61±2.88, 15.15± 2.96, 49.74±3.07, 146.66±17.98, the differences were statistically significant ( t value was 4.466-13.197, P<0.05). The scores of psychological knowledge, skills, social culture and development and the total scores of Transition shock in the experimental group were 30.90±3.15, 19.69±2.74, 32.17±3.06, 103.81±12.24, which were all higher than those in the control group (25.69±2.76, 15.92±3.24, 27.50±4.15, 88.32±11.54), the differences were statistically significant ( t value was 7.237-10.040, P<0.05). Conclusions:competency-based diversified training can effectively improve the bedside nursing comprehensive ability and post competency, reduce their transition shock, and has strong clinical reference significance.