[ ABSTRACT ] AIM: To observe the effect of rapamycin on the apoptosis of mouse astrocytes in vitro.ME-THODS:The astrocytes from C57BL/6J newborn mouse pups were isolated and primarily cultured.The effect of rapamycin on the viability of astrocytes was assessed by MTT assay.The mean fluorescence intensity of SYTOX?Green stain in the astrocytes was detected by fluorescence microplate reader in order to analyze the effects of rapamycin on the cell death in-duced by H2 O2 , ionomycin and/or deferorxamin.DiOC6 (3) staining was used to analyze the mitochondrial membrane po-tential of the astrocytes induced by H2 O2 .Flow cytometry analysis was used to determine the production of ROS in the as-trocytes and mitochondria by staining with H2 DCFDA and MitoSOXTM Red reagent, respectively.RESULTS: Rapamycin at concentration of 0.5 μmol/L protected the astrocytes against cell death induced by H2 O2 or deferoxamine plus ionomy-cin.Rapamycin protected the mitochondrial membrane potential of astrocytes from the injury of H2 O2 .It also reduced the production of ROS in the astrocytes and decreased the level of ROS in the mitochondria.CONCLUSION:Rapamycin re-duces the ROS overload in the mitochondria, keeps mitochondrial membrane potential safety and protects the astrocytes a-gainst apoptosis in vitro.

Objective To explore the relationship between the level of serum Klotho with renal impairment in chronic kidney disease (CKD) patients. Methods 100 patients with CKD 1-5 stages (CKD group) and 30 non-CKD patients (control group) were enrolled. The serum of Klotho was detected by ELISA (enzyme-linked immunosorbent assay ) to investigate the correlation between the level of serum Klotho to progression of CKD. Results The levels of serum Klotho in CKD group 9.81 (7.45,12.2) ng/mL and control group 2.97 (1.75,5.23) ng/mL were significantly different (P < 0.01); The Klotho level declined as renal insufficiency progressed. Pearson correlation analysis showed that serum Klotho in CKD patients was positively correlated with eGFR (estimated glomerular filtration rate), blood calcium and hemoglobin,while was negatively correlated with phosphate, age, logserum creatinine, and logFGF-23. Multiple regression analysis showed age, logScr and logFGF-23 were independent factors for expression of serum Klotho in CKD patients. Conclusion Serum Klotho level in CKD patients decreased with the deterioration in renal function and serum Klotho maybe one of indicators to predict the progression of renal function in CKD patients.

Objective To analyze the specimen types,ward distribution and risk factors for infections caused by extended-spectrum β-lactamase(ESBLs)-producing-Escherichia coli(ECO) in recent two years,so as to provide bacteriological basis for both hospital infection control and clinical anti-infection treatment.Methods Non-repetitive 443 ECO strains isolated from the hospitalized patients in the Third People′s Hospital of Shenzhen were collcted,and the phoenix100 system was employed for bacterial identification and antimicrobial susceptibility tests.ESBLs-ECO was further confirmed by the double-disk synergy test,and the risk factors caused ESBLs-ECO were statistically analyzed.Results A total of 115 strains of ESBLs-ECO were identified among the 443 strains of ECO,which accounted for 26.0%.The ESBLs-ECO strains were mainly isolated from the sputum,urine,and blood specimens.Among the isolated ESBLs-ECO strains,20.9% were isolated from the department of Tuberculosis,13.9% from the department of pediatric,12.2% from the department of live disease,and 8.7% from the department of infection.The male sex,surgery and use of the third generation cephalosporins were independent risk factors of ESBLs-ECO infection.Conclusion The isolation rate of ESBLs-ECO in this hospital is high.It is necessary for the hospital to strengthen the control of nosocomial infections according to the risk factors.More attention should be payed on male patients,the standardization of surgical operation and disinfection,and the restriction of using the third generation cephalosporins,so as to reduce the incidene of ESBLs-ECO infections.

Objective To study the level of serum 25 (OH) D3in children in suzhou area, and to provide scientific basis for the rational supplement of vitamin D for children aged 0-6years.Methods From September2015to September 2016, 15 010children underwent routine physical examination in the Children′s Health Clinic of Suzhou Municipal Hospital were selected, of whom 7 905were male and 7 105were female.The serum 25 (OH) D3was detected by collecting their fingerling blood.Results (1) The mean serum 25 (OH) D3of15 010children aged 0to 6in Suzhou was (35.83±13.23) μg/L, and the mean serum 25 (OH) D3of male and female were (36.48±13.25) and (35.11±13.16) μg/L respectively, and the differences were statistically significant (P＜0.01). (2) The mean level of serum 25 (OH) D3of 0-＜3, 3-＜6, 6-＜12, 12-＜36, 36-＜48and≥48months old children were (34.49±11.53), (41.15±13.86), (48.03±17.25), (46.12±17.69), (28.49±16.55) and (42.28±17.59) μg/L.The detection levels of serum 25 (OH) D3between the age groups were statistically significant (P＜0.05) except the children 3-＜6months and≥48months. (3) From January to December, the detection levels of serum 25 (OH) D3 were statistically significant between different months (P＜0.01) except in January, February, March and November, as well as July and August.The serum25 (OH) D3in each month was graded according to the vitamin D level, and the detection levels of serum 25 (OH) D3between different months were statistically significant (P＜0.01).The proportion of serum 25 (OH) D3over 30μg/L was less than 50％in January, March and November.The ratio ranged from 50％to 60％in February, June and December.The ratio ranged from 60％to 70％in the July, August and September, while the proportion was over 70％in April, May and October.Conclusion The level of serum 25 (OH) D3in children in Suzhou area was decreased obviously, and health education should be strengthened, and attention should be paid to intaking of vitamin D in children.

Objective To evaluate the application value of ultrasound in extracorporeal membrane oxygenation (ECMO)for protecting the liver donation after brain-cardiac death (DBCD).Methods Forty patients with brain death or irreversible brain injury,admitted to Guangzhou General Hospital of Guangzhou Military Command from April 2006 to November 201 4,were eligible for liver donation.The hepatic artery blood flow (QHA),portal vein blood flow (QPV)and ECMO-induced ECMO flow of hepatic artery (VE)of the donor liver were monitored by ultrasound before,5 min after the initiation of ECMO and immediately after ECMO.The changes of total bilirubin (TB),alanine transaminase (ALT)and lactic acid were observed at corresponding time points.Hepatic recovery was subsequently evaluated within 3 months after liver transplantation.Results The mean time of ECMO was (1 .0 ±0.2)h.There was no significant difference in QHA and QPV before and after ECMO (both in P >0.05).And there was no significant difference in liver function parameters before and after ECMO (all in P >0.05).At different time points within postoperative 3 months,the results of ultrasound evaluation and liver function test revealed that the transplant liver function was well recovered in 40 recipients.Conclusions Through monitoring QHA by ultrasound,the best ECMO flow should be chosen,which protects DBCD liver and averts perfusion injury and hypoperfusion.

Objective: To investigate the effect of bilirubin on inflammatory signaling pathway mediated by NOD-like receptor 2(NOD2) in premature infants. Methods: Fifteen cases of premature infants hospitalized at the Department of Neonatology, Children′s Hospital of Soochow University from April 2016 to January 2017, were selected, and 2 mL peripheral blood were collected from 15 cases of premature infants, and the mononuclear cells were isolated and divided into 6 groups, including blank control group (group A), muramyl dipeptide(MDP) group (group B), 102 μmol/L bilirubin group(group C), 102 μmol/L bilirubin+ MDP group (group D), 153 μmol/L bilirubin+ MDP group (group E), 255 μmol/L bilirubin+ MDP group (group F). Group A and group B were stimulated by buffer, group C, group D, group E and group F were stimulated by 102 μmol/L, 102 μmol/L, 153 μmol/L, 255 μmol/L bilirubin, respectively.The supernatant was discarded after 1 h, then the medium was added to group A and group C, and the rest of the 4 groups were agonisted with MDP, the cells were stimulated for 24 h, and then the cells and supernatant fluids were collected respectively, the expression levels of NOD2 mRNA in the cells were determinated by real time-PCR, and the expression levels interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in the supernatant was determinated by enzyme linked immunosorbent assay. Results: The expression levels of NOD2 mRNA had no obvious changes after being stimulated by MDP or by different concentrations of bilirubin(7.16±3.08, 6.19±1.99, 7.02±4.04, 6.84±1.81) compared to those of the blank control group(7.46±3.70)(all P>0.05). After being stimulated by MDP, the expression level of IL-6 (5.13±2.36)was significantly higher than that of the blank control group(3.84±1.44), and the difference was statistically significant(P<0.05), but TNF-α had no obvious changes(4.85±2.47 vs.4.04±2.26, P>0.05). The expression levels of IL-6 and TNF-α had no obvious changes compared to those of the blank control group after being stimulated by 102 μmol/L bilirubin(3.26±1.06 vs.3.84±1.44, 3.45±1.84 vs.4.04±2.26, all P>0.05); but the expression levels of IL-6 and TNF-α were significantly lower after 153 μmol/L and 255 μmol/L bilirubin stimulation (IL-6: 2.58±1.33, 2.16±0.94; TNF-α: 2.32±1.49, 2.42±1.42)compared to those of the blank control group(3.84±1.44, 4.04±2.26), and the differences were statistically significant(all P<0.05). Conclusions Bilirubin have no obvious inhibitory effect on NOD2, and low concentration of bilirubin had no obvious inhibitory effect on IL-6 or TNF-α in this study, but high concentration of bilirubin can inhibit the expression levels of IL-6 and TNF-α, hyperbilirubin may inhibit the ability of anti-infection immune response in body by inhibiting inflammatory signaling pathway mediated by NOD2.

Objective: Toexplore work process-oriented theory nursing ward round, research work process-oriented theory nursing ward round on critical thinking capacity of nursing undergraduates. Methods: Totally 80 Elective nursing ward round courses of nursing undergraduates were divided into the experimental A group and the experimental b group with 40 cases in each group. The experimental A group select the beginning of 9 weeks on Until, the experimental B group select the after of 9 weeks on Until. The nursing undergraduates were assessed by CTDI-CV on first, ninth, eighteenth weeks to evaluate the effect of the two groups. Results: Main effect of group factor and time factor of CTDI-CV had statistical significance (P<0.05) . There were significant interaction in the scores of CTDI-CV. Simple effect analysis demonstrated the scores of CTDI-CV in the experimental A group were higher than those of the experimental b group after the intervention (P<0.05) . Two groups of on first, ninth, eighteenth weeks measured values compared the effect of amount of F_B<F_A, the experimental group A intervention effect was better than in the experimental group B intervention. Conclusions For nursing undergraduates guided based on the working process of the curriculum of nursing ward round the critical thinking ability had good practice guidance, and on the start time to choose university grade three semester two on 9 weeks before commencement of nursing undergraduates promotes higher critical thinking ability.

Objective To compare the effects of different doses of phillyrin(PHN)on the injury of human retinal vascular endothelial cells(RVECs)induced by high glucose(HG)and analyze its regulatory effect on the expression of complement C1q/tumor necrosis factor-related protein-3(CTRP3)and its possible mechanism.Methods RVECs were cultured with HG to establish cell injury models(HG group).RVECs in the HG+PHN-L group,HG+PHN-M group,and HG+PHN-H group were treated with 1 μmol·L-1,10 μmol·L-1,and 100 μmol·L-1 PHN,respectively,followed by HG induction.RVECs in the HG+pcDNA group and HG+pcDNA-CTRP3 group were transfected with pcDNA and pcDNA-CTRP3,respectively,followed by HG induction.RVECs in the HG+PHN-H+sh-NC group and HG+PHN-H+sh-CTRP3 group were transfected with sh-NC and sh-CTRP3,respectively,then induced by HG and treated with 100 μmol·L-1 PHN.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)were meas-ured.Cell apoptosis was detected by flow cytometry.The expression levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax)and CTRP3 protein were determined by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot,respectively.Results Compared with the Con group,the cell apoptosis rate and the levels of MDA and Bax protein in the HG group increased,while the levels of SOD,GSH-Px,Bcl-2 protein,CTRP3 mRNA and protein decreased(all P＜0.05).Compared with the HG group,the cell apoptosis rate and the levels of MDA and Bax protein in the HG+PHN-L,HG+PHN-M,and HG+PHN-H groups decreased(HG+PHN-H group＜HG+PHN-M group＜HG+PHN-L group),while the levels of SOD,GSH-Px,Bcl-2 protein,CTRP3 mRNA and protein increased(HG+PHN-H group＞HG+PHN-M group＞HG+PHN-L group)(all P＜0.05).Compared with the HG+pcDNA group,the cell apoptosis rate and the levels of MDA and Bax protein in the HG+pcDNA-CTRP3 group decreased,while the levels of SOD,GSH-Px,CTRP3 protein,and Bcl-2 protein increased(all P＜0.05).Compared with the HG+PHN-H+sh-NC group,the HG+PHN-H+sh-CTRP3 group showed an increase in the cell apoptosis rate and the levels of MDA and Bax protein and a decrease in the levels of SOD,GSH-Px,CTRP3 protein,and Bcl-2 protein(all P＜0.05).Conclusion PHN can alleviate HG-induced damage to RVECs,which may be related to the upregulation of the CTRP3 expression.

Objective To investigate the risk factors for pulmonary infection in patients with acute-on-chronic liver failure(ACLF),and to establish a predictive model.Methods A retrospective analysis was performed for 585 ACLF patients who were admitted to Department of Infectious Diseases,The Second Affiliated Hospital of Air Force Medical University,from January 2009 to September 2022,and according to the condition of pulmonary infection after admission,they were divided into infection group with 213 patients and non-infection group with 372 patients.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between groups.The clinical data of these patients were collected.Univariate and multivariate Logistic regression analyses were used to investigate the risk factors for pulmonary infection in ACLF patients and establish a predictive model,and the receiver operating characteristic(ROC)curve was plotted to assess the predictive value of the model.The Hosmer-Lemeshow test was used to evaluate the degree of fitting of the model,and the ROC curve and the area under the ROC curve(AUC)were used to assess the predictive performance of the model.Results Among the 585 patients with ACLF,213 experienced pulmonary infection,with an infection rate of 36.41%.The multivariate logistic analysis showed that upper gastrointestinal bleeding(odds ratio[OR]=2.463,P=0.047),infection at other sites(OR=2.218,P=0.004),femoral vein catheterization(OR=2.520,P＜0.001),and combined use of two or more antibiotics(OR=2.969,P＜0.001)were risk factors for pulmonary infection in ACLF patients.These factors were included in the risk factor predictive model of Logit(P)=-1.869+0.901×upper gastrointestinal bleeding+0.755×infection at other sites+0.924×femoral vein catheterization+1.088×combined use of two or more antibiotics.The ROC curve analysis showed that the model had a good predictive value(Hosmer-Lemeshow χ2=3.839,P=0.698),with an AUC of 0.753(95%confidence interval:0.700-0.772).Conclusion There is a relatively high incidence rate of pulmonary infection in patients with ACLF,and upper gastrointestinal bleeding,spontaneous peritonitis,femoral vein catheterization,and combined use of two or more antibiotics are related risk factors.The model established based on these factors can effectively predict the onset of pulmonary infection in ACLF patients.

Objective To establish a quantitative polymerase chain reaction(PCR)method for the analysis of human-derived SRY DNA in mouse tissues,and to study the tissue distribution of human umbilical cord mesenchymal stem cells(HUCMSCs)in immunodeficient NOG mice after a single intravenous injection.Methods We established a quantitative PCR method for the analysis of human SRY DNA in mouse tissues,and validated the standard curve,linear range,accuracy,precision,and stability.Thirty-six NOG mice(18 male,18 female)were administered 3.5×107 HUCMSCs/kg by single intravenous injection.Six mice were then anesthetized and dissected after blood collection(EDTA anticoagulation)at 6,12,24,and 72 h,and at 1 and 2 weeks,respectively.DNA was extracted from lung,kidney,heart,liver,brain,spinal cord,stomach,small intestine,fat,skin,spleen,testis,uterus,and ovary tissues,and the distribution of HUCMSCs in each tissue was determined by the validated quantitative PCR method for detecting the human-derived SRY gene in mouse tissues.In addition,18 NOG mice(9 male,9 female)were divided into control(n = 6)and treatment groups(n = 12)injected intravenously with 0.9%sodium chloride and 3.5×107 cells/kg,respectively.Acute toxic reactions were observed during the administration period,and four animals were dissected at 72 h and at 2 and 4 weeks after administration to observe the gross organs.Mitochondrial protein expression was detected in paraffin sections of lung tissues by immunohistochemistry to analyze the colonization of HUCMSCs in lung tissues.Results The established RT-qPCR method for human-derived SRY DNA in mouse tissues met the validation criteria for each index.After a single intravenous injection in NOG mice,HUCMSCs were mainly distributed in the lungs and blood within 1 week after administration,with higher concentrations in lung tissues than in blood.The concentrations of HUCMSCs in lung tissue and blood remained relatively stable within 6～24 h and 6～72 h,respectively,and then decreased over time.The distribution of HUCMSCs in other tissues was not measured at all sampling points.The colonization result showed that HUCMSCs were detected in lungs 72 h after intravenous injection,but not at 2 and 4 weeks.No obvious acute toxicity was observed in NOG mice after single intravenous administration of HUCMSCs.Conclusions The above method for analyzing the distribution of HUCMSCs in mouse tissue is reliable and feasible.HUCMSCs were mainly distributed in lung and blood in NOG mice within 1 week after a single intravenous injection,and mainly colonized lung tissue at 72 h.A single intravenous administration of HUCMSCs has a good safety profile.