Objective:To investigate the effects of ghrelin on oxidative stress and ferroptosis in retinal microvascular endothelial cells (HRMEC) under high glucose conditions.Methods:HRMEC were divided into control group, high glucose group, high glucose+ ghrelin group and cultured with conventional medium, 30 mmol/L D-glucose medium, and 30 mmol/L D-glucose+ 10 nmol/L ghrelin medium in vitro for 24 hours accordingly.The cell proliferation was identified by cell counting kit-8 assay.The reactive oxygen species (ROS) levels were detected by flow cytometry.The oxidative stress indexes glutathione (GSH) concentration, malondialdehyde (MDA) superoxide dismutase (SOD) activity and Fe 2+ concentration were detected by the corresponding kit.The mitochondrial structure was observed by transmission electron microscopy.The expression levels of glutathione peroxidase 4 (GPX4) and recombinant solute carrier family 7 member 11 (SLC7A11) proteins were detected by Western blot. Results:The cell proliferation rates of control group, high glucose group and high glucose+ ghrelin group were (100.62±3.40)%, (63.74±4.25)% and (88.19±4.65)%, respectively.The ROS fluorescence intensity of control group, high glucose group and high glucose+ ghrelin group was 15 512.20±1 347.53, 46 457.00±1 072.65 and 22 220.87±1 669.20, GSH concentration was (68.52±7.61), (21.45±1.57) and (55.68±5.15)μmol/L, MDA concentration was (0.79±0.10), (2.47±0.27) and (1.08±0.15)μmol/L, SOD activity was (111.67±10.32), (37.75±5.92) and (97.45±9.12)U/ml, Fe 2+ concentration was (3.02±0.30), (9.45±0.71) and (4.63±0.32)mmol/mgprot, respectively.There were statistically significant differences in cell proliferation rate, ROS fluorescence intensity, GSH, MDA, and Fe 2+ concentrations and SOD activity among the three groups ( F=61.82, 414.59, 61.28, 67.24, 61.64, 146.14; all at P<0.001).Compared with the control group, the cell proliferation rate, GSH concentration and SOD activity were reduced, ROS fluorescence intensity, MDA and Fe 2+ concentrations were increased in the high glucose group, with statistically significant differences (all at P<0.05).Compared with the high glucose group, the cell proliferation rate, GSH concentration and SOD activity were significantly increased, ROS fluorescence intensity, MDA and Fe 2+ concentrations were decreased in the high glucose+ ghrelin group, with statistically significant differences (all at P<0.05).Compared with the control group, the ferroptosis of mitochondria in high glucose group was obvious.Compared with the high glucose group, the mitochondrial status of the high glucose+ ghrelin group was significantly improved.There were significantly differences in the relative expression levels of GPX4 and SLC7A11 proteins in cells among the three groups ( F=63.94, 182.84; both at P<0.001).Compared with the control group, the relative expression levels of GPX4 and SLC7A11 proteins were significantly decreased in the high glucose group and high glucose+ ghrelin group (all at P<0.05).Compared with the high glucose group, the relative expression levels of the two proteins in the high glucose+ ghrelin group were increased, with statistically significant differences (both at P<0.01). Conclusions:Ghrelin can promote proliferation of HRMEC under high glucose conditions, and inhibit high glucose-induced oxidative stress and ferroptosis.

Objective To investigate the correlation between random urinary albumin-to-creatinine ratio(ACR)and 24 hour urine total protein quantification(24h UTP)in patients with immunoglobulin A nephropathy(IgAN),and analyze the consistency of these methods in clinical diagnosis.Methods A total of 230 patients with primary IgAN admitted to Peking University Shenzhen Hospital from January 2019 to December 2020 were selected as the research subjects.Correlation analysis and intraclass correlation coefficient(ICC)were used to assess the correlation between ACR and 24h UTP and their consistency in clinical diagnosis.Subgroup analysis was performed using different chronic kidney disease(CKD)stages and urine protein levels.Receiver operating characteristic(ROC)curves were plotted with 24h UTP=0.5 g/24h,1.0 g/24h and 3.5 g/24h as boundary points to determine the optimal cut-off values for ACR.Results There was a positive correlation between ACR[0.79(0.41～1.45)g/g]and 24h UTP[1.02(0.58～1.80)g/24h]in patients with IgAN(r=0.85,P<0.01),and the consistency in clinical diagnosis between the two methods was moderate(ICC=0.63,P<0.01).Subgroup analysis revealed that the correlation and consistency between ACR and 24h UTP was not affected by CKD stages,with correlation coefficients(r)of different CKD staging ranging from 0.76 and 0.86(all P<0.01)and ICC values ranging from 0.53 and 0.72 across different CKD stages.However,it was affected by urine protein level.When 24h UTP was≤0.5 g/24h,there was no significant correlation between ACR and 24h UTP(r=0.08,P>0.05).In subgroups with 24h UTP≤0.5 g/24h,0.5 g/24h<24h UTP≤1 g/24h and 24h UTP>3.5 g/24h,the consistency between the two methods was negligible(all ICC<0.20).ROC curve results showed that when 24h UTP=0.5 g/24h,1.0 g/24h and 3.5 g/24h,the optimal cut-offvalues of ACR were 0.30 g/g,0.57 g/g and 1.28 g/g,respectively.Conclusion In IgAN patients,ACR cannot simply replace 24h UTP for urine protein level evaluation.Especially when 24h UTP≤1 g/24h and 24h UTP>3.5 g/24h,ACR may not accurately reflect the true urine protein level.

Objective:To investigate the risk factors of low anterior resection syndrome (LARS)after low anterior resection of rectal cancer (Dixon).Methods:This retrospective study was conducted in Peking University First Hospital and Traditional Chinese Medicine Hospital of Shanxi Provice from Jan 2012 to Jun 2019. A cohort of 504 patients with rectal cancer was enrolled in the study. All the patients underwent anterior resection. The relationship between clinical-pathological data were analyzed retrospectively. Univariate analysis using χ 2 test. Logistic regression analysis was used to screen the influencing factors of LARS, and the Nomogram method was used to score each factors. Results:Univariate analysis showed that BMI≥28 kg/m 2(χ 2=9.450, P=0.002), the distance from the lower edge of the tumors to the anus <6 cm (χ 2=12.070, P=0.001), high ligation of the inferior mesenteric artery (IMA) (χ 2=8.279, P=0.004), preoperative neoadjuvant therapy (χ 2=11.230, P=0.001), postoperative anastomotic leakage (χ 2=11.840, P=0.001) were associated with severe LARS.Multivariate analysis showed that the distance from the lower edge of the tumors to the anus <6 cm ( OR=1.861, 95% CI: 1.289-2.688, P=0.001), BMI≥28 kg/m 2 ( OR=1.747, 95% CI: 1.022-2.987, P=0.041), high IMA ligation ( OR=1.688, 95% CI: 1.157-2.463, P=0.007), preoperative neoadjuvant therapy ( OR=2.719, 95% CI: 1.343-5.505, P=0.005) were independent risk factors for LARS. Nomogram model showed that the total factor ranged from 2 to 212, and the corresponding risk rate ranged from 30% to 80%. The patients with higher score have greater risk for severe LARS. The area under the predictive power curve of Nomogram model (AUC) was 0.749 (95% CI: 0.705-0.793, P<0.001). Conclusion:Lower tumor location, obesity, preoperative neoadjuvant therapy, high IMA ligation and postoperative anastomotic leakage increase the risk of severe LARS.

OBJECTIVE：To investigate the effects and mechanism of puerarin （Pue） on hypoxia-induced pyroptosis of pulmonary artery smooth muscle cells （PASMCs）. METHODS ：PASMCs of rats as research objects were randomly divided into normoxia group ，hypoxia group and hypoxia+Pue group （0.2 mmol/L）. Except for normoxia group ，other groups were cultured with 5％ CO2 and 3％ O2 at 37 ℃ for 24 hours to establish hypoxia model. Western blot assay was used to detect the expression of pyroptosis related proteins [NOD-like receptor protein- 3 （NLPR3），caspase-1，interleukin-1 β （IL-1 β），apoptosis-associated speck-like protein （ASC）]. Lactate dehydrogenase （LDH）release assay was used to detect the release of LDH in cells ；Hoechst 33342/PI double staining test was adopted to detect the proportion of pyroptosis positive cells. PASMCs was randomly divided into normoxia group+control plasmid group ，hypoxia+control plasmid group ，hypoxia+over-expression plasmid group and hypoxia+ over-expression plasmid+Pue group. Except for the normoxia+control plasmid group ，the other groups were established hypoxia model by the same method. After transfection of control plasmid or NLRP 3 over-expression plasmid ，Western blot ，LDH release test and Hoechst 33342/PI double staining test were used to investigate whether Pue could inhibit hypoxia-induced PASMCs pyroptosis by interfering with the activity of NLRP 3 inflammasomes. RESULTS ：Compared with normoxia group ，the expression of pyroptosis related proteins ，the release of LDH and the proportion of pyroptosis positive cells were increased significantly in hypoxia group （P＜0.05 or P＜0.01）. Pue had the effect of reversing the above indexes （P＜0.05 or P＜0.01）. When the NLRP 3 inflammasome was over-expressed ，the expression of pyroptosis related proteins ，the release of LDH and the proportion of Δ 基金项目：黑龙江省自然科学基金资助项目（No.ZD201416） pyroptosis positive cells were increased significantly （P＜0.05 ＊教授，博士生导师 ，博士。研究方向 ：心血管药理学 。电话： or P＜0.01）. Pue could inhibit the above phenomenon through 0451-58853046。E-mail：zhangxd85@163.com regulating NLRP 3 inflammasome （P＜0.05 or P＜0.01）. 中国药房 2021年第32卷第11期 China Pharmacy 2021Vol. 32 No. 11 ·1337· CONCLUSIONS：Pue can significantly inhibit the hypoxia-induced pyroptosis of PASMCs by down-regulating the expression of pyroptosis related proteins ，reducing the release of LDH and proportion of pyroptosis positive cells. The mechanism is related to the activity inhibition of NLRP 3 inflammasome.

Objective:To investigate the effect of blue light combined with albumin treatment on heart, liver and nerve damage in neonatal jaundice.Methods:A total of 120 cases with neonatal jaundice in the Department of Pediatrics of Women's and Children's Hospital of Zhoushan from April 2017 to April 2018 were selected and divided into control group and observation group accorded to the random number method, with 60 cases in each group.The control group received blue light therapy.The observation group was given albumin treatment on the basis of the control group.The serum total bilirubin, bile acid, aspartate aminotransferase(AST), alanine aminotransferase(ALT), glutamyl transpeptidase(GGT), troponin(cTnT), creatine kinase isozyme(CK-MB), -hydroxybutyrate dehydrogenase(HBDH), myoglobin(MYO), S100B, neuron-specific enolase(NSE) and glial fibrillary acidic protein(GFAP) levels were determined.Results:Before treatment, there were no statistically significant differences in serum total bilirubin, bile acid, AST, ALT, GGT, cTnT, CK-MB, HBDH, MYO, S100B, NSE and GFAP levels between the two groups(all P>0.05). After treatment, the serum levels of total bilirubin[(142.67±13.02)μmol/L, (118.62±11.68)μmol/L], bile acid[(15.34±2.42)μmol/L, (7.83±2.07)μmol/L], AST[(32.17±6.34)U/L, (21.04±5.58)U/L], ALT[(25.83±4.16)U/L, (18.37±4.05)U/L], GGT[(55.24±6.37)U/L, (36.17±5.86)U/L], cTnT[(0.16±0.03)×10 -6μg/L, (0.09±0.02)×10 -6μg/L], CK-MB[(4.32±0.85)×10 -6U/L, (2.01±0.72)×10 -6U/L], HBDH[(213.04±43.61)U/L, (137.26±41.61)U/L], MYO[(22.15±3.64)×10 -6μg/L, (14.26±3.27)×10 -6μg/L], S100B[(1.41±0.28)×10 -9μg/L, (0.87±0.22)×10 -9μg/L], NSE[(15.29±2.12)×10 -9μg/L, (15.29±2.12)×10 -9μg/L] and GFAP[(19.34±0.96)×10 -9μg/L, (14.36±0.92)×10 -9μg/L] in the two groups were lower than those before treatment( t=5.214, 8.261; 7.216, 11.524; 4.027, 6.843; 3.248, 5.764; 7.129, 13.654; 6.524, 9.751; 6.854, 9.031; 4.026, 6.204; 4.521, 7.026; 4.276, 5.846; 4.812, 7.023; 7.062, 13.524, all P<0.05). The levels of serum total bilirubin, bile acid, AST, ALT, GGT, cTnT, CK-MB, HBDH, MYO, S100B, NSE and GFAP in the observation group were lower than those in the control group( t=10.651, 18.267, 10.208, 9.953, 17.066, 15.038, 16.063, 9.738, 12.490, 11.747, 17.157, 29.011, all P<0.05). Conclusion:Blue light combined with albumin treatment can alleviate heart, liver and nerve damage in neonatal jaundice.

Objective:To investigate the use of different tools of intravenous infusion and the issues associated with intravenous catheter indwelling in hospitalized children and to provide reference for clinical practices.Methods:Using the convenient sampling method, from July 24th to 31st, 2018, the intravenous infusion treatment of children inpatients in 49 hospitals within the Beijing Children's Hospital Medical Association was selected for investigation. A self-designed Cross-sectional Questionnaire for Children's Intravenous Infusion and a photo of an intravenous infusion tool were used for data collection. Statistical analysis was performed using SPSS 17.0 software.Results:During the data collection period, there were 18 316 hospitalized children in the 49 hospitals, among which there were 14 421 cases of infusions, with the infusions rate of 78.73%. The application rate of indwelling needle was 87.94%. The connector of venous catheter was mainly heparin cap connection (64.23%) . The infusion tools used in 6 398 cases (44.37%) were made by PVC and containing 2-ethyl hexyl phthalate (DEHP) . Aseptic transparent dressing was the main dressing choice. During catheter indwelling, there were some problems such as back-blood in the pipeline or infusion joint, incorrect clamping position of small clips, unclamping, and tube detachment. Indwelling needles had more problems than central venous catheters, including old blood returning in the tube and blood returning in the infusion connector. The difference between the peripheral indwelling needle and the central catheter was statistically significant ( P<0.05) . Conclusions:The rate of intravenous infusion in hospitalized children is relatively high. Although there are various types of intravenous infusion tools, the choice of infusion tools for children was mainly indwelling needle and heparin cap connection, and there were still room for improvement in catheter maintenance.

Objective To investigate the role and its mechanism of tumor necrosis factor-α (TNF-α) in post-stroke cognitive impairment (PSCI). Methods Sixty male C57BL/6J mice aged 9-11 weeks were randomly divided into sham operation group, PSCI group, and infliximab group. A PSCI model was induced by middle cerebral artery occlusion. The infliximab group was given infliximab intraperitoneally (10 mg/kg, twice a week), and the PSCI group was injected with an equal volume of normal saline. Water maze and light-dark transition tests were used to evaluate cognitive impairment. Western blot analysis was used to detect hippocampal TNF-α and interleukin-18 ( IL-18 ). The levels of kynurenine and tryptophan in hippocampus were measured by high performance liquid chromatography (HPLC), and the changes of indoleamine 2,3-dioxygenase (IDO) activity (the ratio of kynurenine to tryptophan) were evaluated. Results Morris water maze experiment shows that the escape latency of mice was significantly prolonged in the PSCI group, the target quadrant stay time was significantly shortened, and the number of crossing target quadrants was significantly reduced compared with the sham operation group (all P < 0. 05). The escape latency of the infliximab group was significantly shorter than that of the PSCI group, the target quadrant stay time was significantly prolonged, and the number of crossings increased significantly ( all P < 0. 05 ). Light-dark transition test shows that the latency of the mice was significantly shortened and the number of errors was significantly increased in the PSCI group (all P < 0. 05). The latency of the infliximab group was significantly prolonged compared with the PSCI group, and the number of errors was significantly reduced (all P < 0. 05). Compared with the sham operation group, the levels of TNF-α and IL-18 in the mouse hippocampus of the PSCI group were significantly increased (all P < 0. 05), and the kynurenine/tryptophan ratio was significantly increased (P < 0. 05); the level of TNF-α in hippocampus and the ratio of kynurenine/ tryptophan in the infliximab group were significantly lower than those in the PSCI group (all P < 0. 05). Conclusion TNF-α inhibitor infliximab can alleviate PSCI in mice by reducing IDO activity.

Objective To screen the pathogenic gene of osteopetrosis to provide reference for its genetic di-agnosis and prognosis .Methods The clinical data and peripheral blood samples were collected from the pa-tients with osteopetrosis ,DNA was extracted ,the whole exome sequencing library was built ,then the high throughput detection was performed and the pathogenic gene was screened by combining with the bioinformat-ics technology .Results The whole exomes in 2 cases of osteopetrosis were analyzed ,the average sequencing depth of the two samples were 169 .38X and 231 .06X respectively ,in which the case 1 carried rare mutation TCIRG1(c .1305+2T>C) ,TCIRG1(c .2008C> T ) and CLCN7(c .1116C> T );the case 2 carried a rare muta-tion CLCN7(c .857G>A ) .T he bioinformatics analysis indicated that the rare mutations carried by these cases all had different degrees of influence on the structure and function of gene products .Conclusion The whole exome sequencing can once screen the know n pathogenic mutations of osteopetrosis ,is an effective tool for pathogenic mutation screening of osteopetrosis ,the clinical disease in 2 cases of osteopetrosis may be closely related with the patient′s carrying TCIRG1 and CLCN7 mutation .

Objective To investigate the clinical curative effect of Qingrehuatan method in the treatment of insomnia elderly patients with benzodiazepines dependence.Methods From April 2016 to April 2017,49 insomnia elderly patients with benzodiazepines dependence in the Second People's Hospital of Zhoushan were divided into the observation group(20 cases)and control group (29 cases)according to the different treatment methods.In the gradual reduction at the same time,the control group was treated with trazodone,the observation group was treated with Qingrehuatan method.Before treatment and after treatment for 1course,the revised Pittsburgh sleep quality scale (PSQI),index of insomnia symptom score,TCM symptom score,Zung Anxiety Rating Scale score,Wechsler Adult Intelligence Scale(WAIS－RC)and Wechsler Memory Scale(WMS－RC)score were observed in the two groups. Results After treatment,the rate of withdrawal and discontinuation of the observation group was significantly higher than that of the control group (90.0% vs.68.9%),the difference was statistically significant (χ2＝6.918,P＜0.05).After treatment,the PSQI scores and symptom scores of the two groups were significantly decreased,and the PSQI scores and TCM symptom scores of the observation group improved significantly better than those of the control group[(8.8 ±1.1)points vs.(10.6 ±1.3)points,(2.9 ±0.8)points vs.(4.6 ±1.2)points],the differences were statistically significant(t＝5.063,5.535,all P＜0.05).After treatment,the Zung Anxiety Scale scores of the two groups were significantly lower,and the improvement of the observation group was significantly better than that of the control group[(42.90 ±5.65)points vs.(48.21±5.27)points],the difference was statistically significant(t ＝ 3.366,P＜0.05 ).After treatment,the long －term memory,short －term memory and memory quotient of the two groups were significantly increased,and short－term memory and memory quotient of the observation group improved significantly better than those of the control group[(23.30 ±5.31)points vs.(20.55 ±6.58)points,(76.67 ± 12.70)points vs.(67.36 ±19.13)points],the differences were statistically significant (t＝2.274,2.047,all P＜0.05 ).Conclusion Qingrehuatan method in the treatment of insomnia elderly patients with benzodiazepines depend-ence can improve the possible reduction in the process of stopping drug withdrawal symptoms and memory function, and can improve the symptoms of patients at the same time,and it is worthy of clinical application.