Action Potentials ; drug effects ; Animals ; CA1 Region, Hippocampal ; cytology ; Drug Antagonism ; Male ; Neurons ; drug effects ; Pentylenetetrazole ; pharmacology ; Propofol ; pharmacology ; Rats ; Rats, Wistar ; Synaptic Transmission ; drug effects

Objective To establish a brand new model of bladder outflow obstruction (BOO) at single cell level in vitro to offer a more stable and scientific experimental base. Methods Recurrent mechanical stretch generated by vacuum facility was applied to cells attached to flexible membrane of special culture plate, which led to 10%, 20% and 30% elongation of them. Immunocytochemistry was used to analyze the expression of ?-actin (sign of contractile phenotype) and RT-PCR was performed to detect the dynamic changes of mRNA expression of PCNA and Western blotting to protein expression of PCNA and cdk2 kinase. MTT assay was used to observe the changes of proliferation of cells. Results In the case of gradual elongation, the expression of ?-actin rose at first and fell afterwards; the mRNA expression of PCNA and protein expression of PCNA and cdk2 kinase rose gradually. Changes in 30% elongation group were greater than the control (P

Objective To establish a stable animal model of bladder outlet obstruction(BOO) and to clarify the effects of BOO on the biomechanical properties of detrusor and the mechanisms. Methods BOO animal model was established by partial ligation of the proximal urethra of male Wistar rats forming, a urethra stricture with internal diameter of 1 mm. The active contraction of detrusor muscle stripes to carbachol was recorded with transfusor and organ bath tube. The compliance and maximum volume of bladder were examined by filling cystometry. Results The bladders demonstrated tropical post obstruction alterations at week 2 after partial ligation of the proximal urethra. The maximum volume (3 3?1 9) ml and bladder wet weight (10 4?2 8) g increased significantly in BOO group as compared with those in sham operation group [(1 6?0 8)ml and (7 2?3 5)g]. The detrusor contractile force of detrusor instability(DI) group was significantly lower than that in the control group. However, the contractile force in detrusor stability (DS) group was higher at week 2 but lower at week 4 after BOO than that in the sham operation group. Conclusion BOO animal model established by partial ligation of the proximal urethra of adult rats is simple and can give high survival rate and good stability, suitable for the studies of bladder outlet obstruction. There are two types of the detrusor contraction after BOO: DI group with impaired contraction and DS group with bidirectional change which was higher at early stage(week 2) but lower at later stage(week 4). The changes of bladder compliance are related to the stability of bladder, decrease in DI but increase in DS.

Objective To provide experimental basis for further study of the mechanism of urinary storage dysfunction of diabetic cystopathy (DCP). Methods Steptozotocin(STZ) induced and sucrose induced diabetic rats were employed as DCP and diuresis models. Normal rats served as the control. The changes of calcitonin gene related peptide(CGRP) were analyzed by immunocytochemical method and computer assisted image analysis software. Compliance of the whole bladder prepared in vitro was evaluated by bladder irrigation. The messenger molecule cAMP of detrusor relaxation was determined by radioimmunological assay. Results Compared with those in other groups, the CGRP immunoreactive positive nerve fibers and content of this neurotransmitter significantly decreased in the bladder wall, especially in the submucosa. Significantly higher bladder compliance was observed( P

Endodermal Sinus Tumor ; pathology ; Female ; Humans ; Infant ; Vaginal Neoplasms ; pathology

Xiaoyao pills are a famous traditional Chinese medicine collected in Welfare Pharmacy, which is a classic prescription for treating liver depression and spleen deficiency. However, its composition is complex. In order to better control the quality of Xiaoyao pills, in this study, HPLC-ion-trap time-of-flight mass spectrometry (LC-IT-TOF/MS) was used to identify the main ingredients of Xiaoyao pills, paeoniflorin, albiflorin, glycyrrhizic acid, saikosaponin A and saikosaponin B2. Then a liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed for simultaneous determination and quantification of the main compounds. Fragmentation pathways of five active components were obtained. The method was validated. Five active ingredients in Xiaoyao pills had a good linear relationship, and the values of RSD (%) of repeatability were all less than 5%, the recovery ranges were between 90% and 115%, and the values of RSD (%) of each substance were less than 10% after the sample solution is placed for 24 hours. Three batches of Xiaoyao pills (concentrated pellets) and two batches of Xiaoyao pills (water pellets) were determined, the contents of paeoniflorin in concentrated pills were more than 4.0 mg·g^-1, and those in water pills were more than 2.5 mg·g^-1, which was accordance with Chinese Pharmacopoeia. However, other compounds behave differently. This method has high sensitivity and reliable measurement results, which provides basis for quality control of Xiaoyao pills and material basis for pharmacology research.

Objective To explore the characteristics of blood vitamins A, B2, B6, B12, D, E, K1, K2 and folic acid and their correlation with severity in patients with metabolic-related fatty liver disease (MAFLD). Methods　From September to December 2022, a total of 473 cases of residents were recruited through community MAFLD screening activities and their health information was obtained through questionnaire survey and physical examination. The severity of hepatic steatosis was determined with FibroScan, and vitamin concentrations were determined with liquid chromatography-tandem mass spectrometry. Two independent samples' t-tests were used to assess the differences between the two groups, and univariate chi-square tests and multivariate logistic regression analysis were used to explore the related factors of MAFLD. Results　Of the 473 inhabitants, 195 (41.23%, 195/473) met the diagnostic criteria for MAFLD, including mild 43 (22.05%, 43/195) cases of fatty liver, 88 (45.13%, 88/195) cases of moderate fatty liver, and 64 (32.82%, 64/195) cases of severe fatty liver. Using healthy residents collected during the same period as controls, the overall mean of vitamins A, E, K1, and K2 in the MAFLD group was higher than that of the healthy group, with a statistical difference (P<0.05). Furthermore, the concentrations of vitamins A, E, K1 and K2 increased with the severity of fatty liver [R=0.149, P=0.004; R=0.245, P<0.001; R=0.110, P=0.032; R=0.129，P=0.012]. There were statistically significant differences (P<0.05) in the blood levels of vitamin A and E between patients with moderate or severe fatty liver and the healthy population. The blood vitamins K1 and K2 in severe fatty liver patients were also different from those of healthy people (P<0.05). However, there was no significance between folic acid, vitamin D, B2, B6, B12, and MAFLD (P>0.05). Through univariate chi-square analysis and multivariate logistic regression analysis, it was found that male [Wald=5.789, P=0.034，OR=1.598(1.037-2.463)] and vitamin E≥8.13 μg/mL[Wald=14.632，P<0.001，OR=2.378(1.522-3.674)] were risk factors for moderate and severe MAFLD. Conclusions The concentrations of vitamin A, E, and K in the blood are increased in patients with MAFLD compared to the healthy population, and they are positively correlated with the severity of MAFLD. ale gender and high levels of vitamin E may be related to moderate to severe MAFLD.

Carcinoma in Situ ; genetics ; Carcinoma, Transitional Cell ; genetics ; Chromosome Aberrations ; Chromosomes, Human, Pair 17 ; genetics ; Chromosomes, Human, Pair 3 ; genetics ; Chromosomes, Human, Pair 7 ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Urinary Bladder Neoplasms ; genetics

OBJECTIVETo investigate the relationship between phenotype transformation and biomechanical properties of detrusor smooth muscle cell (DSMC) subjected to the cyclic mechanical stretch.

METHODSCultured rat DSMCS were grown on collagen-coated silicone membranes and subjected to continuous cycles of stretch-relaxation. All experiments were made on cells between passage 2 and 4. Each cycle consists of 5-second stretch and 5-second relaxation. The computer controlled vacuum induced 10% (I), 20% (II) and 30% (III) maximum elongation of the plate membrane at different designed pressures. We assessed DNA synthesis rate using tritiated thymidine incorporation assay. Using immunofluorescent assay and flow cytometer, we analysed the expression of SM-alpha-actin and proliferation of DSMC. The image analysis and micropipette aspiration systems were employed to investigate the single cell contraction and viscoelasticity. The elastic modulus K(1), K(2) and viscoelastic coefficient micro were determined using the three-element standard linear solid model, thus demonstrating the passive deformation ability of detrusor cells.

RESULTSAs the basic structural changes to mechanical stretch, DSMCs underwent phenotypic modulation from their normal contractile phenotype to a "synthetic" phenotype: the DSMCs became more proliferative and the actin less organized along the cell's long axis. The cell proliferation index (CPI) of control and stretched group (10%, 20%, 30% elongation) were 0.24, 0.43, 0.58 and 0.65 respectively. After mechanical stretch, the well-spread filaments changed their orientation. Contraction and viscoelasticity of single DSMC subjected to stretch both decreased significantly compared to control. The Vmax and. DeltaLmax of group III (30% elongation) saw significant decreases compared with unstretched control (P < 0.01). K(1) and K(2) decreased with the increasing of mechanical overload, however, there was no statistic difference between groups II and group III.

CONCLUSIONSStructure determines function. Conversely, dysfunction implies the structural transformation. Functional abnormalities of BOO have the structural basis: phenotype transformation of detrusor cells. Cyclic stretch and relaxation applied to DSMCs in vitro can be used to model the increases in urodynamic load experienced by the bladder detrusor muscle under the conditions of bladder outlet obstruction. Phenotypic transformation is the structural basis of functional changes of DSMC subjected to periodic overload mechanical stretch.

Animals ; Biomechanical Phenomena ; DNA ; biosynthesis ; Muscle, Smooth ; physiology ; Phenotype ; Rats ; Rats, Wistar ; Stress, Mechanical ; Urinary Bladder ; physiology ; Urinary Bladder Neck Obstruction ; physiopathology

Objective To evaluate the effect of the intermittent deferomamine(DF) therapy on relieving iron overload caused by transfusion in children with ? thalassemia.Methods Sixteen children who were finally diagnosed as ? thalassemia major were treated with deferomamine for 124 times totally to low the iron overload. The serum iron(SI), serum ferritin(SF) and urine ferritin were detected each time with radio-immunity technique and difference was compared before and after treatment. Meanwhile, weather DF involved children′s liver and renal function was observed in whole procedure.Results Iron overload exists in 16 cases of ? thalassemia major children by a long- term hypertransfusion therapy, with average level SI 33.69?6.72 mmol/L,SF 441.19? 54.70 ?g/L,urine ferritin 8.64?6.79 ?g/L. The difference was significant (paired-samples t test,t =6.173 P 0.05).Conclusion The study suggest that intermittent low-dose DF therapy is effective for iron overload caused by transfusion in ? thalassemia children, without apparent side effects.