Objective To identify whether cisplatin can induce autophagy of bladder cancer T24 cells and the possible mechanism, and to observe the relationship between outophagy and apoptosis.Methods MTT assay was applied to investigate the effects of various concentration of cisplatin( 0 , 10 , 20 and 40 μg/mL) on T24 survival.TEM was performed to detect the autophagosome formation .Western blot assay was used to analyze the expression changes of LC3-Ⅱ, P62 and extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK at the protein level.The effects of autophagy on the survival and apoptosis of bladder cancer cells were investiga-ted.Results DDP observably inhibited proliferation of bladder cancer cells in a dose-dependent manner ( P<0.05), the 50% inhibiting concentration(IC50) was (30.3 ±2.4)μg/mL;DDP induced autophagy of bladder cancer cells, observably increased autophagosome induced by DDP; up-regulated expression levels of LC3-Ⅱproteins ( P<0.05 ) , down-regulated expression of P62 proteins ( P<0.05 );DDP increased the protein level of p-ERK (P<0.05); The inhibitor of ERK pathway U0126 inhibited DDP-induced autophagy, as evidenced by decrease in the expression of LC3-Ⅱproteins ( P<0.05 ) .After inhibition of autophagy by WTM in DDP-treated cells, cell viability was obviously decreased and apoptosis was increased (P<0.05);DDP combined with WTM observably enhanced cleavage of poly ADP-ribose polymerase 1 ( PARP-1 ) and cleaved-caspase-3 which is apop-tosis related proteins ( P<0.05 ) .Conclusions Autophagy can protect T24 cells against ciplatin-induced apop-tosis, the possible mechanism of autophagy is the ERK signaling pathway is activated .

Objective To report two cases of familial Gitelman syndrome and literature review regarding the updates of relevant genes,classification,treatment,and prognosis.Methods The clinical data of two sisters with Gitelman syndrome were retrospectively analyzed.Results Their blood pressures were within normal range.Hypokalaemic alkalosis,hypomagnesemia,and hypocalciuria were corrected almost completely after three days of intravenous magnesium and potassium infusion,spirolactone and indometacin.However,the maintenance of normal potassium was unsuccessful over one year.Conclusion Hypokalaemic alkalosis,hypomagnesemia,and hypocalciuria were normalized in Gitelman syndrome.There was some debate in regard to using PGE2 synthetase inhibitors.Tolerance of long-term medication will be the big challenge for curative effect.

Objective To evaluate the training quality of anaesthesiology residents in different hospitals. Methods The scores of 125 residents,attending Shanghai resident clinic skill examination from 2005 to 2007,were analyzed. ResultsThe scores of examinees from municipal hospitals were higher than those from district hospitals,municipal specialized hospitals and traditional Chinese medicine hospitals.The scores of practice part were higher than those of theory part in all groups. Conclusion The training quality is related to the comprehensive strength of the hospital in which examinees work,but the training of theory knowledge still need to be strengthened.

A new benzene derivative microintegerrin C (1) and a new norsesquiterpenoid microintegerrin D (2), along with six known compounds (3-8), were isolated and identified from stems and leaves of Micromelum integerrimum by various chromatographies such as silica gel, Sephadex LH-20, RP-18 column chromatography and HPLC. Their structures were mainly identified based on the spectral data analysis such as 1D-, 2D-NMR and HR-EI-MS. All known compounds were isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rutaceae ; chemistry ; Sesquiterpenes ; isolation & purification

Objective To evaluate the risk of the occurrence of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer in the oral HPV carriers through a population-based investigation in Shanghai.Methods A total of 1200 cases of outpatients who attended the annual cervical examination and 50 preoperational cases of inpatients with CIN Ⅲ or invasive cervical cancer were enrolled from three clinical centers in Shanghai.The oral HPV infection was determined by real-time PCR.In 1200-case cross-sectional study,the incidence rate of CIN was compared between the oral HPV positive and negative cohort.In 1250-case case-control study,the positive rate of oral HPV DNA test was compared among normal control group,CIN Ⅰ-Ⅲ,and invasive cancer case groups,and all odds ratio (OR) values were calculated,respectively.The HPV transmission-related demographic and behavioral characters of the oral HPV carriers were also analyzed.Results The oral HPV carriers accounted for 5.9％ (71/1200) of the investigated outpatients.The oral HPV DNA positive rates were gradually increased with the cervical disease grades,which were 5.8％ (68/1182,normal),2/13 (CIN Ⅰ),1/5 (CIN Ⅱ),31.4％ (11/35,CIN Ⅲ) and 5/15 (invasive cancer).In cross-sectional cohort studies,the relative risks (RR) of CIN Ⅰ,Ⅱ were 2.9 and 4.0 for oral HPV carriers,respectively.In case-control study,the OR values for CIN Ⅰ-Ⅲ and invasive cervical cancer were 3.1(95％CI:1.6-10.1),4.2(95％CI:1.7-28.4),7.1(95％CI:4.8-19.8) and 10.1 (95％ CI:3.2-32.1),respectively.The oral sex and multi-sexual partner were two major risk factors for the oral and cervical HPV co-infection,HPV-related cervical cancer and precancerous diseases according to behavioral analysis.Conclusions There are complicated transmission pathways between oral and cervicalHPV.Oral HPV carriers should be intensively followed up and their oral HPV infection and HPV-related cervical diseases should be treated together.

Aim To examine the effect of ? opioid receptor agonist U50488H on the levels of interleukin-6(IL-6) and interleukin-8(IL-8) from angiotensinⅡ(AngⅡ) stimulated endothelial cells.Methods In the in vivo study,the modulation of U50488H(1.5 mg?kg~-1) intravenously on the level of AngⅡ was evaluated.In the in vitro study,endothelial cells from human umbilical vein(HUVEC) were cultured and divided into four groups:Control group,AngⅡ group,and AngⅡ plus U50488H and/or nor-BNI(a selective ? opioid receptor antagonist) group.These groups were treated respectively with phosphate buffered solution(PBS),AngⅡ(10~-9~10~-5 mol?L~-1),and AngⅡ in the presence of U50488H and/or nor-BNI for 0~24 hours.Culture supernatant and endothelial cells were collected at 0,3,6,12 and 24 h.IL-6 and IL8 levels in culture supernatant were measured by enzyme linked immunosorbent assay(ELISA).Results The level of AngⅡ in the blood was significantly decreased following U50488H intravenously,which was blocked by nor-BNI administration(2 mg?kg~-1).AngⅡ stimulated the productions of IL-6 and IL-8 from HUVEC in the dose-dependent and time-dependent manners.U50488H at 10~-5 mol?L~-1 significantly inhibited this process,and the inhibitory effect of U50488H was blocked by nor-BNI,which itself had no effect.Conclusion ? opioid receptor may play a role in the process of anti-inflammation via down regulation of AngⅡ level and inhibition of the AngⅡ-stimulated IL-6 and IL-8 productions from endothelium cells.

A series of 4-(2-acetoxybenzoylamino) butyrate derivatives were designed and synthesized. All of the novel 12 compounds (7a-7k) were synthesized from gamma-aminobutyric acid (1) as starting material, and their structures were confirmed with IR, 1H NMR, EI-MS and elemental analysis. Preliminary pharmacological test in vitro showed that most of these title compounds possessed antiepileptic activity. Compounds 7i-7k displayed strong antiepileptic activity and are worth for further development. Compounds 4, 7d-7h showed moderate antiepileptic activity. The structure-activity relationship of 4-(2-acetoxybenzoylamino) butyrate derivatives is also discussed preliminarily.
4-Aminopyridine ; Animals ; Anticonvulsants ; chemical synthesis ; chemistry ; therapeutic use ; Butyrates ; chemical synthesis ; chemistry ; therapeutic use ; Drug Design ; Epilepsy ; chemically induced ; drug therapy ; Female ; Lethal Dose 50 ; Male ; Mice ; Molecular Structure ; Random Allocation ; Structure-Activity Relationship ; gamma-Aminobutyric Acid ; chemistry

Objective To evaluate the accuracy and safety of pedicle-scope assisted pedicle screw fixation and percutaneous pedicle screw fixation for treating lumbar degenerative disease(LDD).Methods Thirty cases of LDD treated by mini-invasive transforaminal lumbar interbody fusion(TLIF) under microscope plus percutaneous pedicle screw fixation with fluoroscopy or pedicle-scope of lumbar spine from December 2013 to September 2015 were selected and divide into the percutaneous group and pediclescope group,15 cases in each group.The operative duration,intraoperative blood loss volume,hospital stay and complications were compared between the two groups.The clinical effects were assessed with visual analogue scale(VAS) and Oswestry disability index(ODI).The plain radiography,dynamic radiology and lumbar magnetic resonance imaging(MRI) and computed tomography (CT) were re-examined after operation for assessing the screw location and fusion rate.Results Eighty-four screws were placed in the percutaneous group and 70 screws in the pedicle-scope group.In the percutaneous group and pedicle-scope group,the mean intraoperative blood loss volume,mean operative duration,average times of C-arm fluoroscopy,mean length of hospital stay,mean time of off-bed and complication occurrence rate were measured,and the differences were not statistically significant (P＞ 0.05).In 6-month follow up,the VAS score and ODI score in the two groups were significantly improved compared with before operation(P＜0.01);the VAS score and ODI score were(2.50 ± 1.30) and (50.00-±-3.50) in pediclescope group,(3.00± 1.50) and (58.00 ±4.50) in percutaneous group,respectivehy,and the differences were statistically significant(P＜0.05);the improvement rates of clinical symptoms in the percutaneous group and pedicle-scope group were 73.33 ％ and 80.00 ％ respectively.The imaging reexamination showed that the placed screw location was good and lesion segment fusion was good without screw loosening.The location of 3 screws in the percutaneous group was deviated and adjusted by the second operation.Conclusion The accuracy and safety of pedicle-scope assisted screw placing for treating LDD are higher than those of conventional percutaneous pedicle screw fixation,moreover the operative time is shorter with less intraoperative bleeding.

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

To investigate the urination-reducing effect and mechanism of Zhuangyao Jianshen Wan (ZYJCW). In this study, SI rats were subcutaneously injected with 150 mg · kg(-1) dose of D-galactose to prepare the sub-acute aging model and randomly divided into the model group, the Suoquan Wan group (1.17 g · kg(-1) · d(-1)), and ZYJCW high, medium and low dose groups (2.39, 1.20, 0.60 g · kg(-1) · d(-1)) , with normal rats in the blank group. They were continuously administered with drugs for eight weeks. The metabolic cage method was adopted to measure the 24 h urine volume and 5 h water load urine volume in rats. The automatic biochemistry analyzer was adopted to detect urine concentrations of Na+, Cl-, K+. The ELISA method was used to determine serum aldosterone (ALD) and antidiuretic hormone (ADH). The changes in P2X1 and P2X3 mRNA expressions in bladder tissues of rats were detected by RT-PCR. According to the results, both ZYJCW high and medium dose groups showed significant down-regulations in 24 h urine volume and 5 h water load urine volume in (P <0.05, P <0.01), declines in Na+ and Cl- concentrations in urine (P <0.01), notable rises in plasma ALD and ADH contents (P <0.05, P <0.01) and remarkable down-regulations in the P2X1 and P2X3 mRNA expressions in bladder tissues (P <0.01). The ZYJCW low dose group revealed obvious reductions in Na+ and Cl- concentrations in urine (P <0.01). The results indicated that ZYJCW may show the urination-reducing effect by down-regulating the P2X1 and P2X3 mRNA expressions in bladder tissues of rats with diuresis caused by kidney deficiency.
Aging ; physiology ; Animals ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; Kidney Diseases ; drug therapy ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X1 ; genetics ; Receptors, Purinergic P2X3 ; genetics ; Urinary Bladder ; metabolism