Objective To compare the expressions of Beclin 1 and its 11 interacting proteins in subcutaneous fat and visceral fat,and to observe the expressions of these genes in 3T3-L1 adipocytes differentiation process.Methods In 3T3-L1 adipocytes,cells were harvested at day 0,2,4,6,8 in differentiation process,and then proteins or mRNAs were obtained and followed with Western blot and realtime PCR.Mice tissue proteins or mRNAs were detected with Western blot and realtime PCR.Results In 3T3-L1 adipocytes differentiation process,expressions of LC3,Beclin 1,and TNFAIP3 continued to rise,expression of BIRC5 was high to low,FEZ1 continued at a low level.In mice,expressions of LC3,Beclin 1,YWHAQ,FEZI,BAD,WAC,TNFAIP3,and GOPC in visceral fat were significantly higher than those in subcutaneous fat,while expression of SLAMF1was higher in subcutaneous fat.Conclusion Autophagy plays an important role in 3T3-L1 differentiation process and may be tissue-specific in visceral fat and subcutaneous fat.

Objective To clone human autoimmune antigen SSA/Ro60 and to purify its expression to provide the material basis for the assisted diagnosis of human autoimmune diseases .Methods The SSA/Ro60 gene was cloned by RT‐PCR and directionally inserted into expression vector pPICZ .The recombinant plasmid was transformed into Pichia SMD1168 .The obtained recombinant protein was identified by SDS‐PAGE and Western blotting .Results The amplified full‐length sequence was about 1 .5 kb in size . The pPICZ‐SSA positive clone produced a 60 × 103 recombinant protein which had natural immunogenicity of human autoimmune antigen SSA/Ro60 by SDS‐PAGE and Western blot .Conclusion Human autoimmune antigen SSA/Ro60 is successfully cloned and expressed ,which lays a foundation for diagnosing autoimmune diseases .

Objective To evaluate the effects of sinomenine on the expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) during renal ischemia-reperfusion (I/ R) in rats.Methods Fifty-four male Wistar rats, weighing 180-220 g, were randomly divided into 3 groups (n=18 each) using a random number table: sham operation group (group S) , group I/R and sinomenine group (group SIN).Renal I/R was induced by clamping the left renal pedicle for 45 min followed by reperfusion in I/R and SIN groups.In group S, the bilateral renal pedicels were only exposed.Sinomenine 60 mg/kg was intraperitoneally injected at 30 min before reperfusion in group SIN, while the equal volume of normal saline was given in group S and group I/R.At 6, 12 and 24 h (T1-3) of reperfusion, 6 rats from each group were chosen, and blood samples were drawn from the hearts for determination of the serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The animals were then sacrificed, and the left kidney was removed and embedded into a paraffin block for determination of the expression of HIF-1α and VEGF in the renal tissues (by immuno-histochemistry).Results Compared with group S, the serum Cr and BUN concentrations were significantly increased at T1-3 , and the expression of HIF-1α was significantly up-regulated at T2,3 in group I/R and group SIN, and the expression of VEGF was significantly upregulated at T2,3in group I/R, and at T1-3 in group SIN.Compared with group I/R, the serum Cr concentration at T1-3 and serum BUN concentration at T2,3 were significantly decreased and the expression of HIF-1α at T2,3and VEGF at T1-3was significantly up-regulated in group SIN.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to up-regulation of the expression of HIF-1α and VEGF in the renal tissues of rats.

Objective To investigate the effect of sinomenine on the levels of nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-two healthy male Wistar rats, aged 6-9 weeks, weighing 180-220 g, were randomly assigned into 4 groups (n =18 each) using a random number table: control group (group C), sinomenine group (group SIN), group I/R, and sinomenine+I/R group (group SIN+I/R).The animals were anesthetized with 10％ chloral hydrate 350 mg/kg.The left renal pedicles were clamped with atraumatic microclips for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion to establish the model of renal I/R injury.Sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion in group SIN +I/R, and at the corresponding time point in group SIN.At 6, 8 and 12 h of reperfusion, the blood samples were drawn by cardiac puncture for measurement of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The left renal specimens were obtained for examination of pathological changes (with light microscopes) and for determination of the rate of NF-κB-positive cells and iNOS expression in renal tissues (by immunohistochemistry).Results Compared with group C, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly increased, and the expression of iNOS was up-regulated in I/R and SIN+I/R groups, and no significant change was found in the parameters mentioned above in group SIN.Compared with group I/R, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly decreased, and the expression of iNOS was down-regulated in group SIN+I/R.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN+I/R compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activity of NF-κB and down-regulated expression of iNOS in rats.

Objective To evaluate the effect of sinomenine on apoptosis in renal tubular epithelial cells of rats subjected to renal ischemia-reperfusion (I/R), and the relationship with C-Jun N-terminal kinase (JNK) signaling pathway.Methods Fifty-four male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were randomly divided into 3 groups (n =18 each) using a random number table: sham operation group (group S), I/R group and sinomenine group (group SIN).Renal ischemia was induced by occlusion of the left renal pedicle for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion in anesthetized rats in I/R and SIN groups.In group SIN, sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion, while the equal volume of normal saline was given instead of sinomenine at the same time point in S and I/R groups.Six animals in each group were selected at 0.5, 6 and 24 h of reperfusion, blood samples were collected by cardiac puncture for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.Immediately after blood sampling, the left kidney was removed for examination of pathological changes in renal tissues (with light microscopes) and for determination of phosphorylated JNK (p-JNK) and caspase-3 expression (by immune-histochemistry) and apoptosis in renal tubular epithelial cells (by TUNEL).The apoptotic rate was calculated.Results Compared with group S, the serum Cr and BUN concentrations were significantly increased, the expression of p-JNK and caspase-3 was up-regulated, and the apoptotic rate was increased in I/R and SIN groups.Compared with group I/R, the serum Cr and BUN concentrations were significantly decreased, the expression of p-JNK and caspase-3 was down-regulated, and the apoptotic rate was decreased in group SIN.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activation of p-JNK signaling pathway and reduced apoptosis in renal tubular epithelial cells of rats.

Objective To evaluate the effect of lappaconitine on renal ischemia-reperfusion (I/R) injury in mice.Methods Thirty-six male Wistar rats,weighing 180-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and lappaconitine group (group LA).Renal I/R was induced by occlusion of the left renal pedicle for 45 min with atraumatic microclips followed by reperfusion,and the right kidney was removed after atraumatic microclips were released.At 30 min before reperfusion,lappaconitine 4 mg/kg was injected intraperitoneally in group LA,and normal saline 2 ml was given in S and I/R groups.In group S,the left renal pedicle was only isolated.At 5 and 24 h of reperfusion,blood samples were taken from the inferior vena cava for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations,and kidney specimens were obtained for histopathologic examination (with light microscope) and for determination of the expression of cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) in renal tissues (by immunohistochemistry).Results Compared with group S,the serum Cr and BUN concentrations were significantly increased,and the expression of COX-2 and MMP-2 in renal tissues was up-regulated at 5 and 24 h of reperfusion in I/R and LA groups.Compared with group I/R,the serum Cr and BUN concentrations were significantly decreased,the expression of COX-2 and MMP-2 in renal tissues was down-regulated at 5 and 24 h of reperfusion and histopathologic changes were reduced in group LA.Conclusion Lappaconitine can attenuate renal I/R injury through inhibiting the expression of COX-2 and MMP-2 in rats.

Objective:By analyzing the clinical characteristics of anti-Ku antibody associated disease, this paper aims to improve the diagnosis and treatment level of it.Methods:The clinical symptoms, laboratory tests and prognosis of 40 anti-Ku-antibody positive patients from the First Affiliated Hospital of Zhengzhou University from September 2017 to September 2019 were retrospectively collected, and then hierarchical clustering analyzed.Results:The average age of 40 anti-Ku positive patients was 48±18 years, and the male to female ratio was 1∶4. The average follow-up was (11±7) months, of which 2 cases were accompanied by malignant tumors and 3 cases died. Interstitial lung disease was most common and was found in 24 cases (60%). The most common disease was inflammatory myopathy (11 cases, 28%), followed by systemic lupus erythematosus (SLE) (9 cases, 22%). According to hierarchical cluster analysis, the anti-Ku-antibodies positive patients were divided into 3 groups. Among them, group A had the highest incidence of pulmonary interstitial fibrosis (84%, P<0.01), and the lowest incidence of renal involvement (0, P<0.01), cytopenia (0, P<0.01), serositis (0, P<0.01). Although the incidence of anti-Jo-1 antibody positivity in group A was the highest (16%, P=0.44) but without statistically significant difference. The characters of group A were in line with inflammatory myopathy. Group C had the highest incidence of renal involvement (57%), lupus rash (71%), cytopenia (57%), low complement (71%) and lupus-related antibodies positivity ( P value were all<0.05), which was in line with SLE. These two groups had their own significant biological characteristics, and were rarely overlapped. Conclusion:Anti-Ku antibody appears in a wide spectrum of autoimmune diseases, among which inflammatory myopathy is the most common, followed by SLE. Patients with anti-Ku antibody rarely have SLE and myositis overlapped, and the overall prognosis is good, but it is necessary to be alert to complications, such as tumors.

Objective To evaluate the effect of community management of diabetic patients with hypertension in Beijing Cuigezhuang community in last three years.Methods A community diabetic management program was started from 2007 in Beijing Cuigezhuang community.Three hundred and seventy six patients who participated in the program for more than 3 years were enrolled in the study, including 196 with type 2 diabetes mellitus (T2DM) only (DM group) and 180 with T2DM and hypertension (DMH). The control rate of blood glucose, blood pressure, lipids and the comprehensive control rate were compared between two groups after 3-year intervention.Results There were no significant differences in age, gender ratio, course of diabetes, education background, monthly income and the history of stoke between two groups;while prevalence of dyslipidemia in DMH group was significantly higher than that in DM group [41.7%(75/180) vs.24.5%(48 196),χ2 =11.938,P=0.001].Compared with the baseline data, the types of antidiabetic drugs used were not significantly changed in two groups after 3-year intervention ( DM group:1.32 ±0.81 vs.1.31 ±0.93, t=-0.155, P=0.877, DMH group:1.43 ±0.72 vs.1.48 ±0.82, t=0,831, P =0.407) .The types of antihypertensive drug in DMH group were significantly decrease. (1.12 ±0.77 vs.1.25 ±0.45, t=2.484, P=0.014), while the rate of statins usage in DM group was significantly increased [13.3%(26/196) vs.5.1%(10/196),χ2 =7.830, P=0.005].The hemoglobin A1c (HbA1c) levels in DM group was decreased [(7.4 ±1.5)% vs.(7.8 ±2.1)%, t=2.586, P=0.011].The systolic pressure [(129 ±12) mmHg (1 mmHg=0.133 kPa) vs.(133 ±16) mmHg, t=3.503, P=0.001] and the diastolic pressure [(80 ±8) mmHg ratio (82 ±10) mmHg, t=2.436, P=0.016] in DMH group were significantly declined. The average LDL-C level [ DM group: ( 3.0 ± 0.9) mmol/L vs.(3.2 ±1.0) mmol/L, t =2.165, P=0.032; DMH group (2.9 ±1.0) mmol/L vs. (3.2 ±1.1) mmol/L, t=3.210, P=0.002] were also significantly decrease.Compared with the baseline, the comprehensive control rates of blood glucose, blood pressure and lipid level were increased in both groups [DM group:9.7% (19/196) vs.6.1%(12/196),χ2 =1.716, P=0.190, DMH group 13.9%(25/180) vs.5.0%(9/180),χ2 =8.315, P =0.004] .Conclusions The community management program is effective for improvement of comprehensive control rates of blood glucose, blood pressure and blood lipids in diabetic patients with hypertension in Beijing Cuigezhuang community.

Objective: To determine the influencing factors of electronic screen time of urban preschoolers before and after the COVID-19 outbreak, so as to provide a scientific basis for the control of digital screen use and early prevention of myopia among preschoolers. Methods: Using multi stage cluster random sampling method, a cross sectional survey of 8 244 kindergarten students in a district of Shanghai was implemented, through parent questionnaire collecting the time child spent on various electronic screens before and after the COVID-19 outbreak, estimated the weighting screen time, and emphatically analyzed the relationship between family electronic screen supervision behavior and preschoolers weighting screen time. Results: The proportion of daily over use time on average of mobile phones, computers and TV/projection screens among the surveyed preschool children during COVID-19 was 30.52%, 51.40% and 56.82%, respectively. On school days before the epidemic, the proportion was 21.94%, 41.80% and 47.51% respectively. After controlling for primary covariates, parents frequent control of children s electronic screen use, parents guidance for electronic screen use were significantly associated with lower weighted screen refractive time ( OR =0.60-0.77, P < 0.05 ). The use of electronic screen when parents accompanied their children, the use of electronic screen time by parents but not strictly implemented were significantly associated with higher weighted screen refractive time and increased screen refractive time ( OR =1.18-1.80, P <0.05). Conclusion Urban preschoolers electronic screen time was high during and before COVID-19. In the control measures of preschool children s electronic screen time, attention should be paid to the management of electronic screen use within the family and parents role model.

Objective: The current situation and associated factors of primary and middle school students screen time and related eye hygiene in different periods before and after COVID-19 were investigated, so as to provide a scientific basis for taking targeted measures of preventing and controlling myopia among those students. Methods: A total of 11 402 students were selected from 24 primary schools, 18 junior high schools and 16 high schools by multi stage stratified cluster sampling method. And a questionnaire survey was conducted among students and their parents to understand students screen time and screen hygiene behaviors and related factors in different periods before and after the outbreak of COVID-19. Results: The rate of screen overuse was 7.26%, 9.12% and 35.30% during school days, weekends and home study, respectively. The overuse rate of mobile phones and tablets at home increases with the increase of school stage. Totally 16.30% of students use mobile phones for online learning. The screen overuse rate of those who have not been diagnosed with myopia was lower than that of those who have been diagnosed during home study( OR=0.77, P <0.05). The higher the parents educational level was, the lower the screen overuse rate of children during the weekends became( P <0.05). In any period, the high proportion of screens used in homework and poor home screen eye environment increased the risk of screen overuse and poor eye hygiene ( P <0.05). Parental good behavior was conducive to reducing the risk( OR= 0.65-0.97, P <0.05). When the family had regulation on the screen use time, the implementation of laxity would increase the risk of screen overuse ( OR=1.18-2.48, P <0.05). Conclusion The screen overuse rate of urban primary and middle school students during home study is high, and a certain proportion of students still use mobile phones for online learning. The proportion of screens used in homework, home screen eye environment and screen use management are related to screens overuse. More attention should be paid to those with myopia and low educational level of parents.