Objective To compare the advantages and disadvantages of two methods for rat overactive bladder(OAB)model construction in?duced by intraperitoneal injection and intravesical instillation cyclophosphamide. Methods A total of 30 female SD rats weighting 200?250 g were randomly categorized into three groups:intraperitoneal injection?induced OAB(Ip?OAB),intravesical instillation?induced group(Iv?OAB)and con?trol group. Ip?OAB rats was i.p. administrated cycbophosphe mide three times in dose of 75 mg/kg body weight,while Iv?OAB rats received intravesi?cal instillation three times in drug dose of 75 mg/kg body weight. Control group rats received no treatment. Maximum bladder capacity(MBC),maxi?mum voiding pressure(MVP),frequency of spontaneous contraction of each group were recorded. The incidence,the mortality and the pathology of the three groups were compared. Results MBC,MVP and frequency of spontaneous contraction between Ip?OAB group and Iv?OAB group had no statistically significant difference(P>0.05). Compared with the control group,MBC significantly increased(P<0.05),MVP significantly decreased (P<0.05),and frequency of spontaneous contraction significantly increased(P<0.05)in Ip?OAB and Iv?OAB rats. The modeling success rate and mortality were 100%and 80%in Ip?OAB group,and were 50%and 0%in Iv?OAB group,and pathological changes were found in the two groups. Conclusion The construction of experimental animal model of OAB in rat induced by intraperitoneal injection and intravesical instillation cyclo?phosphamide are both reliable methods. Ip?OAB rats exhibit high incidence and mortality rate,while Iv?OAB rats show low incidence and mortality rate.

AIM: To assess the effect of raloxifene, a selective estrogen modulator, on the transcriptional activity of estrogen responsive element (ERE) in vascular endothelial cells. METHODS: Vascular endothelial cells were transfected with ERE-TK-Luc reporter plasmid and PRL-SV40 control plasmid via FuGENE 6. The activities of firefly luciferase and renilla luciferase were measured with dual-luciferase reporter assay system. The transcriptional activity in transfected cells treated with raloxifene was compared with that in untreated cells. Furthermore, raloxifene was used to cotreat the cells with estradiol (E_2) and the influence of raloxifene to the effect of E_2 was assessed. RESULTS: Compared to untreated cells, the luciferase activity in cells treated with raloxifene decreased and showed significance at concentration of 10~(-7)mol/L (P

Objective Sarcopenia and metabolic syndrome share similar pathophysiological mechanisms. The aim of this study was to investigate the prevalence of sarcopenia among health examination population, and to analyze the relationship between sar-copenia and blood pressure, blood glucose, uric acid and lipids. Methods Physical examination data of 1191 healthy persons in the medical examination center of the hospital from Mar 2011 to Jun 2011 were collected. The weight, skeletal muscle, body fat, body mass index ( BMI) , waist circumference,body fat percentage, waist-hip ratio and visceral fat area were analyzed by human body compositionanalyzer and the prevalence of sarcopenia was observed. At the same time, triglyceride (TG), total cholesterol (TC), high density lipo-protein-cholesterol ( HDL-C ) , low density lipoprotein-cholesterol ( LDL-C) , uric acid and fasting blood glucose were also detected. Results The prevalence rate of sarcopenia of the subjects was 5.21%, and the highest incidence was found in ≥60 years group( 11.11%) . The prevalence rates of overweight and obesity were 33.8% and 10.2%, respectively. The prevalence of sarcopenia is grad-ually higher along with increasing BMI. The prevalence rates of sarcopenia of overweight and obesity subjects were 5.47% and 26.23%, respectively. Compared with the normal control group, the level of weight[(66.34±11.75)kg vs (76.71±12.84)kg ], BMI[(23.37± 3.13) vs (28.05±3.66)], body fat percentage[(25.33±6.06)% vs (36.76±4.47)%], waist circumference[(83.19±9.56)cm vs (95.45±13.74)cm] and visceral fat area[(88.96±29.74)cm2 vs (136.91±25.56)cm2] were higher in the sarcopenia group (P<0.05). Compared with the normal control group, the incidence of systolic blood pressure[(125.59±30.04)mmHg vs (139.39±19.79) mmHg], diastolic blood pressure[(75.82±11.95)mmHg vs (82.34±10.96)mmHg ] TG[(1.56±1.12)mmol/L vs (1.98±1.72)mmol/L] and uric acid[(313.75±83.07)mmol/L vs (335.55±96.07)mmol/L] were higher in the sarcopenia group (P<0.05). Compared with the normal subjects, the detectable rates of abnormal diastolic blood pressure, fasting blood glucose, uric acid, and LDL-C were increased in the sarcopenia, obesity and sarcopenia combined with obesity subjects (P<0.05). The odds ratio of abnormal systolic blood pressure, diastolic blood pressure, uric acid, and LDL-C increased in the sarcopenia, obesity and sarcopenia combined with obe-sity subjects using logistic regression analyses after correction of gender and age. Conclusion The sarcopenia may have some con-nection with metabolic risk factors. Early detection of sarcopenia can help to distinguish people predisposed to metabolic syndrome, and it has important significance for prevention of chronic disease.

Objective To evaluate and analyze the characteristics of cognitive and memory disorders in patients suffering cerebral infarction. Methods The scores of overall cognitive function including orientation to time and place, immediate memory, calculation, delayed memory, naming, repeat, understanding, reading, writing, visuospatial function were evaluated by mini-mental state estimate (MMSE) and the scores of memory function including immediate learning and memory , short time delayed recall , long time delay memories were evaluated by WHO/UCLA Auditory Verbal Learning Test. Statistical analysis were done in patients with cerebral infarction and healthy controls. Results The scores of overall cognitive function, memory scores as well as immediate recall , short-delayed recall , long-delayed recall in cerebral infarction patients were much lower than those of healthy controls (P < 0.001). Conclusion The overall cognitive function and memory function declined significantly , also immediate memory and short term memory were remarkable impaired in patients with cerebral infarction, resulting their memory characteristics were similar to patients with Alzheimer′s disease.

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

Objective To optimize the extraction methods of mitochondrial genome DNA(mtDNA)of Oncomelania hupen-sis. Methods The pyrolysis,protein K variable-temperature digestion and high-concentration potassium acetate purification were applied to optimize the high-concentration-salt precipitation method,and then the optimized method was compared with two common extraction methods,the sucrose density gradient centrifugation method and traditional high-concentration-salt pre-cipitation method. The mtDNA samples were identified by using spectrophotometry,agarose gel electrophoresis and the amplifi-cation products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. Results The concen-tration and yield of the improved method was significantly higher than those of the traditional method(F=3032.65,10185.00, both P<0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein,meeting PCR,sequence analy-sis and other molecular biology research requirements. Conclusions The improved high-concentration-salt precipitation meth-od for isolating mtDNA is simple,and it has high yield and low cost. The extracted mtDNA can meet relevant analysis require-ments.

OBJECTIVELow or moderate consumption of red wine has a greater benefit than the consumption of other beverages in the prevention of atherosclerosis and coronary heart disease and this is increasingly attributed to the polyphenol compounds in red wine, such as resveratrol. In the present study, we investigated the effect of resveratrol on platelet aggregation in vitro and in vivo.

METHODSPlatelet aggregation in rabbits and normal subjects was measured using Born's method.

RESULTSResveratrol, at 10 - 1000 micromol/L, significantly inhibited platelet aggregation in vitro induced by collagen, thrombin, and ADP in healthy subjects. The inhibitory effect was concentration-dependent. Hypercholesterolemia induced by high-cholesterol diet enhanced ADP-induced platelet aggregation. Resveratrol 4 mg x kg(-1) x d(-1) inhibited ADP-induced platelet aggregation in vivo despite no changes in serum lipid levels.

CONCLUSIONSResveratrol inhibits platelet aggregation both in vitro and in vivo. This may be one of the mechanisms by which resveratrol prevents atherosclerosis.

Animals ; Arteriosclerosis ; prevention & control ; Cholesterol, LDL ; blood ; Humans ; Lipids ; blood ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rabbits ; Stilbenes ; pharmacology

In order to study the efficiency of small interfering RNA (siRNA) transfer mediated by cationic liposome, we used luciferase siRNA to evaluate the gene silencing activity in the Hep-2 cells, which were stably transduced with a luciferase gene. The pDNA transfection was studied, and siRNA arrearage assay was conducted to determine the capability of cationic liposome with siRNA. Different concentrations of siRNA was used to silence luciferase gene' activity, and then the result was examined by microplate reader. Cell viability was analyzed after transfection by MTT assay. The results suggested that Lipofectamine 2000 could transfer the pDNA efficiently, and have strong binding capacity with siRNA. The silencing efficiency of luciferase was obtained with low concentration of siRNA. The cell viability was influenced by RNA interference (RNAi) very slightly, but the cell survival rate decreased with the increase of siRNA concentrations. It was well concluded that by optimizing the experimental conditions, cationic liposome can transfer low concentration siRNA to silence target gene's activity efficiently.
Cations ; chemistry ; Drug Carriers ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lipids ; chemistry ; Liposomes ; chemistry ; Luciferases ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

Objective:To construct a primary health care institutions performance evaluation index system from the perspective of health value orientation under the background of countywide medical alliances construction.Methods:From May 2021 to February 2022, preliminary screening was made on core performance evaluation indexes via literature review; purposive sampling was used to select the dean/vice dean, persons in charge of medical service, and those in charge of public health service responsible for performance evaluation at the community health service center. Then semi-structured interviews were made on the existing performance evaluation and assessment plans as well as existing problems of primary medical and health institutions. Based on the " input-process-output" performance evaluation model, the thematic framework analysis method was used to analyze the interview data, and combined with literature research results, a preliminary performance evaluation index system for primary medical and health institutions was built under the guidance of health value. From March to May 2022, the Delphi expert consultation method was used to evaluate the importance and operability of indexes. The threshold method was used to screen indexes, and analytic hierarchy process was used to calculate the weights of evaluation indexes.Results:The health value oriented performance evaluation index system for primary healthcare institutions included 3 first-level indexes, 9 second-level indexes, and 50 third-level indexes. The first-level indexes were output (0.377 3), input (0.336 3), and process (0.286 4) in descending order of weight. The top three weighted second-level indexes were health manpower(0.177 8), health literacy and health outcomes (0.157 6), as well as responsiveness and satisfaction (0.142 6). The third-level indexes included 17 medical indexes, 16 prevention indexes, and 17 medical prevention integration indexes. The top three weighted indexes for inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and responsiveness of residents seeking medical treatment (0.040 1); The top three weighted third-level indexes excluding inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and surplus funds used for personnel incentives (0.045 5).Conclusions:The performance evaluation index system of primary health care institutions built under the health value orientation is scientific, conducive to promoting the health-orientated transformation and improving the efficiency of primary health care services.

Objective To investigate clinical effects and prognostic factors of transabdominal laparoscopic-assisted and open radical resection for Siewert type Ⅱ and Ⅲ adenocarcinoma of the esophagogastric junction (AEG).Methods The retrospective cohort and case-control studies were conducted.The clinicopathologieal data of 84 patients with Siewert type Ⅱ and Ⅲ AEG who were admitted to the First Affiliated Hospital of Xiamen University from January 2014 to January 2017 were collected.Among 84 patients,42 undergoing transabdominal laparoscopic-assisted radical gastectomy (LARG) were allocated into LARG group and 42 undergoing transabdominal open radical gastectomy (ORG) were allocated into ORG group.Observation indicators:(1) comparison of intraoperative and postoperative recovery situations between groups;(2)comparison of follow-up and survival situations between groups;(3) prognostic analysis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival situations,tumor recurrence and metastasis of patients up to January 2018.Measurement data with normal distribution were representde as-x±s,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the ehi-square test or Fisher exact probability.Ordinal data were compared between groups using the Mann-Whitney U nonparametric test.The survival rate and curve were respectively calculated and drawn by the Kaplan-Meier method,and Log-rank test was used for survival and univariate analyses.Multivariate analysis was done by COX proportional hazard model.Results (1) Comparison of intraoperative and postoperative recovery situations between groups:patients with Siewert type Ⅱ and Ⅲ AEG in the LARG and ORG group underwent successful transabdominal radical resection,without conversion to thoracotomy.All patients in the LARG group underwent esophagojejunostomy with circular stapler device,38 and 4 patients in the ORG group underwent esophagojejunostomy with circular stapler and linear cut stapler respectively.Operation time,volume of intraoperative blood loss,length of incision,time of postoperative analgesia,cases with anastomotic bleeding,anastomotic leakage,abdominal bleeding,incisional infection,pulmonary infection,abdominal infection and reflux esophagitis of grade Ⅰ ～ Ⅱ postoperative complications and duration of postoperative stay were respectively (261±50)minutes,(119±111)mL,(7.8±1.6)cm,(2.1±1.3)days,1,1,0,0,1,0,0,(12.8 ± 1.9) days in LARG group and (216 ± 52) minutes,(230± 178) mL,(17.3± 1.8) cm,(3.4±1.2)days,2,0,2,2,2,1,2,(18.4±15.3)days in ORG group,with statistically significances between groups (t =2.357,2.960,2.195,2.013,x2 =5.486,t =2.125,P＜0.05).All patients with complications were improved by symptomatic treatment.(2) Comparison of follow-up and survival situations between groups:81 of 84patients including 41 in LARG group and 40 in ORG group were followed up for 6-48 months,with a median time of 29 months.The postoperative 2-year overall and tumor-free survival rates were respectively 85.1％ and 82.1％ of 41 patients in LARG group and 83.1％ and 79.3％ of 40 patients in ORG group,with no statistically significance between groups (x2 =0.013,0.049,P＞0.05).(3) Prognostic analysis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection:results of univariate analysis showed that tumor diameter,tumor TNM staging,tumor T staging,tumor N staging and postoperative adjuvant chemotherapy were related factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection (x2 =8.349,14.376,9.732,17.250,8.012,P＜0.05).Results of multivariate analysis showed that tumor TNM staging and postoperative adjuvant chemotherapy were independent factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection (risk ratio =4.305,0.031,95％ confidence interval:1.858-9.977,0.004-0.246,P＜0.05).Conclusions Transabdominal laparoscopicassisted radical resection for AEG is safe and feasible,with advantage of minimally invasiveness,having equivalent long-term effects compared to open surgery.Tumor TNM staging and postoperative chemotherapy are independent factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection.