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.

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 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.

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 analyze clinical characteristics and genetic characteristics of children with ATP sensitive potassium passage (K ATP-HI). Methods:Forty-five children with genetically confirmed K ATP-HI and their families admitted to Beijing Children′s Hospital of Capital Medical University between February 2002 and December 2018 were selected as the study subjects. A detailed retrospective analysis of the patient's clinical characteristics, diagnosis and treatment process, disease-causing gene carrying status and later follow-up data was performed. ABCC8/KCNJ11 gene was sequenced by second-generation sequencing technology. Results:Among 45 children with K ATP-HI, 34 cases (75.6%) were neonatal onset, the first symptoms of 21 cases (46.7%) were convulsions. 39 cases had been treated with diazoxide, including 12 cases (30.8%) with good efficacy, 16 cases (41%) with poor efficacy and 11 cases with uncertain efficacy. Octreotide was further applied in 18 patients with uncertain or ineffective efficacy after diazoxide treatment, and 13 cases (72.2%) were effective, 3 cases were ineffective, and 2 cases were uncertain. 10 CHI patients who were ineffective to drug treatment or had clearly focal lesions confirmed by 18F-dopa positron emission by computed tomography ( 18F-DOPA PET) scans had undergone surgical treatment, 8 of which underwent partial pancreatectomy and blood glucose returned to normal after the operation; the other 2 cases underwent subtotal pancreatectomy and both had secondary diabetes after operation. Among 45 children with K ATP-HI, 1 case carried both ABCC8 and KCNJ11 mutations, 10 cases carried ABCC8 compound heterozygous mutations, and the remaining 34 cases carried ABCC8/KCNJ11 single genetic mutation. Among them, 21 cases had paternal inheritance, and 3 cases had maternal inheritance, 6 cases were identified with de novo mutations. Conclusions:Diazoxide treatment was ineffective for most K ATP-HI children, but octreotide had a higher effective rate. Partial pancreatectomy for focal type patients had a higher cure rate, and there was a risk of secondary diabetes after subproximal pancreatectomy, so it was very important to clarify the histological type of children before surgery. ABCC8 gene mutations and KCNJ11 gene mutations were the main pathogenic genes of K ATP-HI. Among patients carrying mutations in single ABCC8 or KCNJ11 gene mutation, K ATP-HI inherited by paternity were the majority. Some K ATP-HI children can relieve the hypoglycemia symptoms by themselves.

Objective: To analyze the status of parents sharing meal expenses in the national pilot areas of the Nutrition Improvement Program for Rural Compulsory Education Students (NIPRCES) and its influencing factors, so as to provide scientific data for promoting the proper implementation of the NIPRCES and the healthy development of students. Methods: In the 726 national pilot counties in 22 provinces where the NIPRCES was implemented, 8 109 primary and junior high schools were selected as the survey objects and included in the analysis according to the three feeding methods of canteen feeding, enterprise feeding and mixed feeding. Data such as the basic information of the schools and counties, and the status of parents sharing meal expenses were collected through questionnaires. Multivariate Logistic regression analysis was used to analyze the influencing factors of parents sharing meal expenses. Results: In the national pilot areas of the NIPRCES, schools where parents shared meal costs accounted for 30.2% of all monitored schools in 2019. In central and western areas, the proportion of schools where parents shared meal costs were 51.5% and 20.3%, respectively. In schools where parents shared meal costs, no matter in primary school or junior high school, the difference in the amount of meal expenses shared by parents of students in middle and western schools was statistically significant ( χ 2= 181.78 , 89.54, P <0.01). Among 38.7% of the parents of primary school students in the central region shared meal expenses of ≥2 yuan, and 47.5% of the parents of primary school students in the western region. Multivariate Logistic regression analysis showed that the parents of schools in the central region, junior high schools or nine year system schools, canteen feeding or other feeding, large school size, the nutritious meal subsidy used for lunch, middle and high family income level, medium and low input level of the monitoring counties, monitoring counties with other nutrition public welfare projects and insufficient working funds share a higher proportion of meal expenses ( OR=3.22, 1.51, 1.36, 1.74, 3.74, 1.38, 11.58, 2.40, 3.15, 1.50, 2.12, 3.36, 1.34, P < 0.05). Conclusion The proportion of meals shared by parents of school students in the NIPRCES implementation area was relatively low, which may be affected by factors such as economic level, feeding mode, school scale, etc.