OBJECTIVE:To prepare silybin liposomes(SLBL)coated by N-trimethyl chitosan(TMC)(TMC60-SLBL),and to optimize the formula. METHODS:The effects of 3 kinds of preparation methods on encapsulation efficiency of SLBL were com-pared,including film dispersion method,multiple emulsion method and reverse evaporation method. The formula of TMC60-SLBL was optimized by orthogonal test using encapsulation efficiency as index with the concentration of phospholipid,ratio of phospholip-id to cholesterol,ratio of silybin to lipids,hydration temperature as factors. The stability of TMC60-SLBL by optimal formula at 4 and 25℃within 30 d,the particle size and Zeta potential of TMC60-SLBL and SLBL were all compared. RESULTS:The encapsu-lation efficiency of TMC60-SLBL prepared by the film dispersion method was the highest,without statistical significance in encap-sulation efficiency before and after cutting (P>0.05). The optimal formula was with the concentration of phospholipid 6 mg/ml;the ratio of phospholipid to cholesterol 40∶1;the ratio of silybin to lipids 1∶30;the temperature of hydration medium 45 ℃;the encapsulation efficiency were(82.08±2.6)%,RSD=3.17%(n=3). The preparation was stable at 4℃;the mean diameter of SL-BL and TMC60-SLBL were(131.9±1.9)nm and(161.2±2.0)nm,and Zeta potential respectively were(-23.18±1.14)mV and (36.73 ± 2.84) mV. CONCLUSIONS:TMC60-SLBL is prepared successfully,and its formula is simple and practicable with high encapsulation efficiency.

Objective To explore the control effect of proactive monitoring on senile stroke women with Escherichia coli infection caused by indwelling catheter.Methods From October 2011 to September 2012 elderly female stroke patients with indwelling urinary catheter were chosen in the department of neurology and were given proactive monitoring,which included the following:management of antibiotics has been humanized,urinary catheter care used first-cleaning and then disinfection method,pathogen monitoring professionals directly went to the bacteria room for obtaining information.The monitoring results were compared with those last year.Results The urinary Escherichia coli infection in monitoring popu-lations declined sharply,Escherichia coli infection rate reduced from 20.5％ to 3.6％.The urinary infection rate in hospitalized patients reduced from 10.8％ to 5.7％ respectively.The preventive application of an-tibacterial drugs greatly reduced from 70.0％ to 24.7％.The nursing staff all mastered the nursing method of the catheter.Conclusions To give proactive monitoring to senile stroke women with indwelling urinary catheter can control Escherichia coli and other bacteria induced urinary infection due to indwelling catheter significantly,demonstrate a deterrent effect on unreasonable application of antibiotics,which can be recommended to control other indwelling catheter induced urinary hospital infection.

Objective:To study the effect of Levocarnitine preconditionon on the myocardial ultrastructure of ischemic rats and the long last calcium channel. Methods: We randomly divided 30 rats into a control group, a Levocarnitine group and a Dihydrochloride group. After the establishment of ischemic models, we observed the ultrastructure of the ischemic myocardium with optical and transmission electron microscopes, and detected the influx of calcium in the long last calcium channel with the patch clamp. Results: Levocarnitine protected the ultrastructure of ischemic myocardium and inhibited the influx of calcium in the long last calcium channel. Conclusion: Levocarnitine can reduce the severity and extent of ischemia-induced damage to the myocardium, protect mitochondria, stabilize its oxidation, inhibit the long last calcium channel of ischemic ventricular myocytes and lessen the injury induced by calcium overload.

Objective:To explore the value of lactic acid (Lac), procalcitonin (PCT), sequential organ failure assessment (SOFA) score and acute physiology and chronic health evaluationⅡ (APACHEⅡ) score in assessing the severity and predicting the prognosis in sepsis shock.Methods:A retrospectively study was conducted. Patients with septic shock hospitalized in the department of critical care medicine of the Affiliated Hospital of Jining Medical University from April 2015 to June 2019 were enrrolled. The patient's gender, age, body mass index (BMI), infection site, organ dysfunction status; Lac, PCT, C-reactive protein (CRP), heart rate and body temperature immediately after admission to the intensive care unit (ICU); APACHEⅡ and SOFA scores within 24 hours, and 28-day prognosis were collected. According to the 28-day prognosis, the patients with septic shock were divided into the survival group and the death group, and the differences in the indicators between the groups were compared. Multivariate Logistic regression analysis was used to screen the risk factors of 28-day death in patients with septic shock; receiver operating characteristic curve (ROC curve) was used to analyze the value of Lac, PCT, SOFA, APACHEⅡ, and age in predicting the 28-day prognosis of patients with septic shock.Results:A total of 303 septic shock patients were enrolled, of which 124 cases survived and 179 died on the 28th day, and the 28-day mortality was 59.08%. ① Compared with the survival group, patients in the death group were older (years old: 66.58±15.22 vs. 61.15±15.68), APACHEⅡ, SOFA, proportion of lung infections, Lac increased [APACHEⅡ score: 22.79±7.62 vs. 17.98±6.88, SOFA score: 9.42±3.51 vs. 5.65±1.59, proportion of lung infections: 53.63% (96/179) vs. 39.52% (49/124), Lac (mmol/L): 5.10±3.72 vs. 3.71±2.56], oxygenation index (PaO 2/FiO 2) and ICU body temperature decreased [PaO 2/FiO 2 (mmHg, 1 mmHg = 0.133 kPa): 198.94±80.15 vs. 220.68±72.06, ICU body temperature (℃): 37.47±1.08 vs. 37.80±1.14], and the differences were statistically significant (all P < 0.05).②Multivariate Logistic regression analysis results: after adjusted for potential confounding factors, APACHEⅡ, PCT, Lac, age and SOFA were independent risk factors for death in patients with septic shock [APACHEⅡ: odds ratio ( OR) =1.05, 95% confidence interval (95% CI) was 1.01-1.10, P = 0.039; PCT: OR = 0.99, 95% CI was 0.98-1.00, P =0.012; Lac: OR = 1.23, 95% CI was 1.08-1.40, P = 0.002; age: OR = 1.03, 95% CI was 1.01-1.05, P =0.009; SOFA score: OR =1.88, 95% CI was 1.59-2.22, P < 0.001]. ③ROC curve analysis showed that APACHEⅡ, Lac, age and SOFA could predict the prognosis of patients with septic shock [APACHEⅡ: the area under the ROC curve (AUC) = 0.682 4, 95% CI was 0.621 7-0.743 1, P = 0.000; when the best cut-off value was 18.500, its sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio were 72.63%, 54.84%, 69.89%, 58.12%, 1.608 1 and 0.499 2, respectively. Lac: AUC = 0.604 5, 95% CI was 0.540 8-0.668 2, P = 0.002; when the best cut-off value was 3.550 mmol/L, the sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio were 50.84%, 73.39%, 73.39%, 50.94%, 1.910 3 and 0.669 9, respectively. Age: AUC = 0.599 1, 95% CI was 0.535 4-0.662 7, P = 0.003; when the best cut-off value was 72.500 years old, the sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio were 42.46%, 75.00%, 71.03%, 47.45%, 1.698 3 and 0.767 2, respectively. SOFA: AUC =0.822 3, 95% CI was 0.776 7-0.867 9, P = 0.000; when the best cut-off value was 7.500, its sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio were 68.72%, 87.90%, 89.13%, 66.06%, 5.680 4, 0.355 9 respectively]. The combined prediction had a good sensitivity (72.63%) and specificity (84.86%), and AUC (0.876 5) was higher than that of a single variable, suggested that the multivariate combination was more accurate in predicting the short-term outcome of septic shock. Conclusion:Lac, PCT, SOFA score, APACHEⅡ score and age were independent risk factors for death in patients with septic shock, and the accuracy of Lac, SOFA score, APACHEⅡ score and age in predicting short-term prognosis of septic shock was better than that of single variable, and the diagnostic value was higher.

Objective To investigate the expression of inducible co-stimulator (ICOS) and inducible co-stimulator ligand (ICOSL) on PBMCs,and the plasma concentrations of soluble forms of ICOSL and their clinical relationship with systemic lupus erythematosus (SLE) patients.Methods Peripheral blood samples were collected from 45 SLE patients and 39 healthy subjects (HC).The expressions of ICOS and ICOSL on peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry.The concentrations of soluble ICOSL were assessed by measured by enzyme linked immunosorbent assay (ELISA).And the relationship between their expression levels and patients' clinical manifestations were analyzed.Levene F test was used for statistical analysis,the comparison between groups was conducted using t test,and the correlation between two variables were tested by Pearson correlation analysis.Results The expression of ICOS on CD4+ and CD8+ T cells were significantly higher than that of the HC [(19.1±1.7)％ vs (14.0±1.5)％,t=2.156,P=0.035],[(10.0± 1.0)％ vs (6.4±1.0)％,t=2.587,P=0.012].The expression of ICOSL on CD14+ mononuclear cells in SLE patients was significantly higher than that in the HC group [(2.94±0.88)％ vs (0.89 ±0.21)％,t=2.152,P=0.04].Plasma ICOSL concentrations in patients with active SLE were significantly higher than those of patients with inactive SLE [(362±25) ng/ml vs (278±15) ng/ml,t=2.356,P=0.025].We also found a significant negative correlation between the soluble ICOSL expression and the surface ICOSL expression on both mononuclear cells and B cells (r=-0.4243,P=0.022;r=-0.4099,P=0.025).MMPI induced an evident reduction in sICOSL levels released from the cells,which was statistically significant in comparison with untreated cells (P＜0.05).Conclusion The up-regulated expressions of ICOS and ICOSL on peripheral lymphocytes and the high levels of plasma concentration of soluble ICOSL are closely correlated with the severity of the disease,suggesting that ICOS/ICOSL pathway may play a critical role in the pathogenesis of SLE.

Objective To investigate the relationship between apoptosis in epidermal keratinocytes and efficacy of epidermal grafting. Methods Epidermal specimens were obtained from donor sites and depigmented area of 44 patients with vitiligo receiving epidermal grafting. The apoptosis in keratinocytes was determined by terminal deoxynucleotidyl transferase end-labeling (TUNEL) assay, and the expressions of caspase 3, 8, 9 as well as bcl-2 and P53 were evaluated by immunohistochemistry. Results As TUNEL assay showed, the number of apoptotic keratinocytes in epidermis from depigmented area differed significantly from that from donor sites.The expressions of caspase 3, 8 and 9 were mainly located in the membrane and cytoplasm of keratinocytes,and positive keratinocytes were predominately distributed in the middle and lower layer of the epidermis. Of the 44 patients, 19, 15 and 16 were positive for the expressions of caspase 3, 8 and 9 in the depigmented epidermis,respectively, and 9, 5 and 4 for those in the donor epidermis, respectively. P53 was expressed in neither donor epidermis nor depigmented epidermis, while Bcl-2 was weakly positive in donor epidermis and negative in depigmented epidermis. The number of apoptotic keratinocytes was higher in donor epidermis from patients failing to respond to the transplantation than in that from patients successfully treated by transplantation (15.83 ± 2.69 vs.9.24 ± 1.80, t = 10.96, P < 0.01 ). Conclusions There is an obvious apoptosis in keratinocytes from depigmented epidermis of patients with vitiligo, together with an increase in the expression of caspase 3, 8 and 9. The apoptosis in keratinocytes may be related to the efficacy of epidermal transplantation.

Objective To study the changes of circulating triglyceride (TG) in the obese elderly, and to investigate the effect of hypertriglyceridemia on the development of insulin resistance. Methods A total of 82 subjects were divided into simple obesity group, obesity with IGT group, obesity with T2DM group and normal control group. The body height, body weight, blood pressure, fasting plasma glucose (FPG) and insulin(FPI), circulating TG and total cholesterol (TC) were measured. The homeostasis model assessment-insulin resistance (HOMA-IR) was calculated. Results The levels of serum TG were significantly higher in the simple obesity group ( 1.3 ± 0. 6) mmol/L, obesity with IGT group (1.9 ± 0.9)mmol/L and obesity with T2DM group (2.1±0.7) mmol/L than in the normal controls [(1.0±0.2)mmol/L, all P<0. 05], Furthermore, the levels of TG, FPG, FPI, HOMA-IR and systolic pressure were significantly higher(all P<0. 05) in obesity with IGT group and obesity with T2DM group than in simple obesity group. There were significantly positive correlations between TG and body mass index (BMI), FPG, HOMA-IR, TC, systolic pressure respectively (all P < 0.05). Multiple linear regression analysis indicated that FPG and HOMA-IR were the independent factors affecting TG (both P < 0. 05 ). Conclusions Hypertriglyceridemia may play an important role in the development of insulin resistance.

Objective:To investigate the expression of inducible costimulatory ( ICOS) and inducible costimulatory ligand ( ICOSL) on peripheral blood mononuclear cells ( PBMCs ) and their clinical relationship with rheumatoid arthritis ( RA ) patients.Methods:Peripheral blood samples were collected from 85 RA patients and 50 HC in this study.Expression of ICOS and ICOSL on PBMC from the subjects were detected by flow cytometry and real-time polymerase chain reaction( RT-PCR).The alteration of ICOS and ICOSL were observed after hormone therapy in 15 patients with RA and the relationship between their expression level and patients′clinical manifestations were analysed.Results:The ICOS and ICOSL mRNA level of RA patients′PBMCs were significantly higher than that in HC.The expression level of ICOS on CD4+T cells was higher than than that in HC[(7.08±4.72)% vs (3.01+1.39)%,P<0.0001].The expression of ICOSL on monocytes[(5.77±3.45)%vs (3.64±1.43)%,P<0.05] and B cells [(5.78± 4.52)%vs (3.97±1.63)%,P<0.05] were significantly elevated in RA patients.In RA patients with active disease,however,ICOSL expression on monocytes and B cells were increased as compared with those in inactive RA patients [ ( 5.45 ±3.50 )% vs ( 4.04 ± 1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Furthermore,after receiving immunosuppressive therapy, the expressions of ICOS and ICOSL were notably reduced as compared with pre-therapy levels on PBMCs from patients [ ( 5.45 ±3.50)%vs (4.04±1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Conclusion:The high levels of ICOS and ICOSL expression were closely correlated with the degree of disease and therapeutic response,suggesting that ICOS/ICOSL pathway may play a critical role in pathogenesis of RA.

Objective To explore serum caspase-3 activity in pulmonary tuberculosis patients.Methods 38 tuberculosis pa-tients in Infectious Disease Department in the First Affiliated Hospital of Guangxi Medical University were recruited and the peripheral venous blood was collected in December,2010.Among them,sputum smear-positive patients which were first treated were 13,sputum smear-negative patients which were first treated were 15,10 cases were drug resistance.In addition, 22 unrelated healthy individuals from a routine health survey were enrolled as controls.Spectrophotometry was used to de-tect serum caspase-3 activity in tuberculosis patients,which was absorbancy-A of 4-nitroaniline generated by caspase-3.Re-sults The activity of caspase-3 in sputum smear-positive patients,sputum smear-negative patients,drug resistance patients, and healthy controls were 37.15±5.06,50.38±6.29,45.32±3.32 and 23.89±2.29,respectively.The caspase-3 activity was significantly different from each other (F=10.68,P<0.01).Compared with the healthy controls,the caspase-3 activity in sputum smear-positive patients (q=3.51,P=0.016),sputum smear-negative patients (q=7.5,P<0.001),drug resist-ance patients (q=5.34,P=0.001)was significantly different.There was no statistical difference among sputum smear-posi-tive patients,sputum smear-negative patients and drug resistance pulmonary tuberculosis patients (P>0.05).In the same way,there was no significant difference in male/female ratio[χ2=0.38 (DL=3),P=0.94]and mean age (F=0.82,P=0.49)among the four group.Conclusion Caspase-3 closely associated with the development of tuberculosis occurrence, which might promote apoptosis of cells through the activation of caspase-3 and produce the body immune process of tubercu-losis.

Objective To measure the level of androgen receptor (AR) mRNA in peripheral blood monocytes (PBMCs) and serum testosterone level of patients with gouty arthritis (GA) and healthy controls (HC),and to explore the role of testosterone and AR in the pathogenesis of GA.Methods Chemilluminescence was used to detect the level of serum testosterone in GA [including 119 acute GA (AGA) and 60 nonacute GA (NAGA) patients] and 47 HC group.Real-time quantitative polymerase chain reaction (RT-qPCR)was used to measure AR mRNA in PBMCs from 41 GA and 35 HC.Western blotting was used to measure PBMCs AR in GA and HC for each 6 cases.One-way ANOVA,t test and Spearman's correlation were adopted for statistical analysis.Results Serum testosterone was significantly reduced in AGA and NAGA group compared to that in HC group [(6.1±1.5) ng/ml,P＜0.01,respectively],and the expression was lower in the AGA [(3.7±1.4) ng/ml] group [(4.9±2.0) ng/ml] than that in the NAGA group (P＜0.01).The level of AR mRNA and protein was much lower in the GA group than that in the HC group (P＜0.01,respectively).Negative correlations was detected between AR mRNA and uric acid in GA patients.There was negative correlation between serum testosterone and VLDL,GLU; meanwhile,positive correlation was found between serum testosterone and HDL (P＜0.05,respectively) in NAGA patients.There were no correlations between testosterone and other laboratory data.There was no correlation between AR and other laboratory data in GA patients and healthy controls (P＞0.05,respectively).Conclusion Altered expression of testosterone and its receptor may be involved in the pathogenesis of gouty inflammation.Further study will be needed to shed light on the exact role of androgen and AR in gout.