AIM:To investigate the effects of extracellular cysteine/cystine redox potential (EhCys/CySS) on the mitochondrial function of nonalcoholic fatty liver disease ( NAFLD) hepatocytes.METHODS:LO2 cells were incuba-ted with EhCys/CySS of the oxidized (0 mV), the normal (-80 mV), or the reduced (-150 mV) status medium, then treated with oleic acid to establish NAFLD model in vitro.DCFH-DA and MitoSOX were used as the fluorescent probes for determining reactive oxygen species (ROS).Apocynin (NADPH oxidase inhibitor), MitoQ10 (mitochondria-targeted an-tioxidant), rotenone (mitochondrial respiratory chain complex I inhibitor) and antimycin A (mitochondrial respiratory chain complex III inhibitor) were used to investigate the sources of ROS.RESULTS:An increase in ROS in LO2 cells by oleic acid was aggravated by the oxidized extracellular EhCys/CySS (0 mV), which was removed by the reduced EhCys/CySS (-150 mV) .ROS generation by 0 mV was significantly eliminated by MitoQ10 .ROS levels were dependent on ex-tracellular Eh Cys/CySS in rotenone treated LO2 cells.A decline of mitochondrial membrane potential in the cells with NAFLD was aggravated by 0 mV and reversed by -150 mV.CONCLUSION:The oxidized extracellular Eh Cys/CySS via inhibitiing of complex I intensifies ROS generation and reducing the mitochondrial membrane potential in the NAFLD hepa-tocytes, which were reversed by reduced Eh Cys/CySS.

This article concerned the idea and working theory of YL-Ⅱ computer's hardware circuit for diet therapy along with the idea of system software, application software and some flowcharts. Thereby, the nutrition specialist system was implemented in a system centred in one slice including.A.The composition of YL-Ⅱ computer for diet therapy.B.The design of YL-Ⅱ nutrition medical treatment computer's hardware circuits for diet therapy. C.The design of YL-Ⅱ computer's software for diet therapy.D.The design of YL-Ⅱ computer's application software for diet therapy. There were 4 main functions of the YL-Ⅱcomputer used in clinic;A.Offering a variety of diet menu; 9 kinds and 84 types of diet menu were stored in the computer, and could be used according to the change of patient's content and the actual intake of the patient.B.Nutritional assessment; To give nutritional assessment with patient's height, weight, skinfold thickness and biochemical parameters etc.C.Nuturitional advisory; the nutritional guide for individual or group.D.Nutritional inquiry.The YL-Ⅱ nutrition medical treatment computer had been used in the clinic of 50 hospitals, all the staffs acknowledged its effectiveness, usefulness, accuracy and time saving, and raising working efficiency 25 times.

To establish a sensitive and specific method for simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS, plasma samples were extracted and derivatized before injection. An ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring (MRM). Norgestrel was chosen as internal standard and performed on a C18 (100 mm x 2.1 mm, 5 microm) column. The concentrations of gestodene, etonogestrel and ethinylestradiol were measured, using step-gradient mobile phase and step-gradient flow rate. The method was validated over the concentration range of 0.1-20 ng x mL(-1) for gestodene and etonogestrel and 0.01-2 ng x mL(-1) for ethinylestradiol, and showed excellent linearity. The intra- and inter-assay accuracy and precision were below 10.0% and recovery was 93.6%-110.9% over the three concentration levels evaluated. The method was applied in pharmacokinetic study of the compound gestodene patch and the compound etonogestrel patch in rabbits. The LC-MS/MS method was selective, accurate and sensitive, especially the LOQ were 100 pg x mL(-1) for gestodene and etonogestrel and 10 pg x mL(-1) for ethinylestradiol. The method was successfully applied in pharmacokinetic study for contraceptives.

Aim To observe the modulation of adenosine triphosphate (ATP) on motility of isolated gastric smooth muscle from rat.Methods Longitudinal and circular muscle strips of rat gastric body and antrum were prepared, and the effects of ATP (0.1, 1,10,100 ?mol?L~-1 and 1 mmol?L~-1 ) on the motility of the strips were investigated. Results ATP induced a small relaxation followed by an obvious spasm in longi- tudinal muscle strips of the gastric body; when tone was raised by KCl, ATP induced a larger relaxation followed by a smaller spasm.ATP only produced contraction in circular muscle strips of the gastric body.Low concentration of ATP inhibited the contractile amplitude of the antral longitudinal strips, at the same time increased the frequency of the contraction apparently; when tone was raised by KCl, ATP produced a concentration-related relaxation.Low concentration of ATP firstly enhanced and then inhibited contractile amplitude of antral circular musle strips, while high concentration of ATP fully inhibited the motility. Conclusion We firstly reported the obvious effects of ATP on longitudinal and circular strips of rat gastric body and antrum. The effects of ATP on the four preparations were different from each other, suggesting that ATP plays an important modulatory role on motility of gastric smooth muscle of rats.

AIM: To study the effect and mechanism of chlorophyllin (CHL) inhibiting HT29 cells. METHODS: IC 50 value and growth curve of HT29 cells were detected with MTT method. Apoptosis was detected with Wright-Giemsa staining, FCM and DNA electrophoresis. Telomerase was detected by PCR-ELISA, and protein and mRNA expression of COX-2 gene were detected through RT-PCR and Western blot. RESULTS: CHL inhibited the growth of HT29 in a dose-dependent manner. CHL blocked HT29 cells in G 1 phase but did not induce apoptosis. Different concentration of CHL inhibits the expression of telomerase and COX-2 in HT29 cells. CONCLUSION: CHL inhibits the growth of HT29 cells by inhibiting the expression of telomerase and COX-2 and blocking cells in G 1 phase. [

Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.

Objective To evaluate the clinical significance of high frequency ultrasonography in the diagnosis of annular pancreas in neonates.Methods Ultrasound results,clinical data,operation results and complicating deformity of 19 neonates were reviewed retrospectively.The digestive tracts of 19 neonates,including stomach,duodenum,jejunoileum and colon,were examined with 8-12 MHz linear transducer before surgery.Results Of 19 neonates with annular pancreas,17 cases were diagnosed with ultrasound,the diagnostic rate was 89％.The pancreas tissue encircling the descending duodenum was directly displayed as a direct sign of ultrasonography and the expansion of the proximal end of the duodenum was presented as an indirect sign of ultrasonography in 17 neonates.There were 3 neonates with annular pancreas complicating intestine malrotation and 2 neonates with annular pancreas complicating duodenal membraneous stenosis.One neonate was misdiagnosised and the other was missed.Conclusions High frequency ultrasonography plays an important role in diagnosing annular pancreas and other digestive deformity in neonates.It can be used as the first choice for the neonatal annular pancreas.

Objective To investigate effects of xeroderma pigmentosum B(XPB) gene on IL-6 induced proliferation and apoptosis in human vascular smooth muscle cells (VSMC).Methods Recombinant plasmid pcDNA3.1-XPB and vacant vector plasmid pcDNA3.1 were transfected stably into VSMC by liposome,and these cells were incubated with IL-6 at a 100 U/ml for 48 hours.The experiments were divided into six groups:blank control group; pcDNA3.1 group; pcDNA3.1-XPB group;IL-6 group; IL-6 + pcDNA3.1 group; IL-6 + pcDNA3.1-XPB group.The expression levels of XPB,Bcl-2,Bax and wild type p53 (wt-p53) were detected through reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.The cell survival,cell cycle and apoptosis were examined with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry,respectively.Results The transfection of pcDNA3.1-XPB increased the expression of XPB,Bax and wt-p53 (P＜0.05 or P＜0.01),decreased the expression of Bcl-2 (P＜0.05 or P＜0.01),and reduced the IL-6 induced effects on decreasing the expression of Bax and wt-p53 and increasing the expression of Bcl-2(P＜0.05 or P＜0.01).The over expression of XPB inhibited the cell growth(q=2.95,P＜ 0.05),and reduced the positive effects of IL-6 on VSMC growth ( 102.6 +6.2) ％ vs.(124.5 + 7.9) ％,q=3.49,P＜0.05.The over expression of XPB increased the apoptosis rate of VSMC(P＜0.01 ) and the cell amounts of G0/G1 phase (q=2.99,P＜ 0.05),decreased the cell amounts of S phase(q=3.05,P＜0.05),and reduced the IL-6 induced effects on decreasing the apoptosis rate of VSMC(5.9±2.1)％ vs.(0.3±0.1)％,q=7.53,P＜0.01; the cell amounts of G0/G1 phase(70.9±6.7) ％ vs.(54.8±2.9) ％,q=6.91,P＜0.01 ;and on increasing the cell amounts of Sphase(20.2+3.6)％ vs.(36.4+7.2)％,q=8.54,P＜0.01.Conclusions XPB gene could inhibit VSMC proliferation,promote VSMC apoptosis,and reduce the effects that IL-6 promotes VSMC proliferation and inhibits VSMC apoptosis.Therefore,XPB gene is likely to be potential molecular target for treatment of atherosclerosis.

Objective To investigate prevention and treatment of cerebrospinal fluid leakage (CSFL)in anterior cervical spine surgery for severe ossification of posterior longitudinal ligament (OPLL).Methods A retrospective analysis of 47 patients with severe cervical OPLL (thickness of the ossified mass ＞ 5 mm,spinal stenosis ＞50％),who had undergone anterior cervical surgery between January 2008 and May 2011,was conducted.Fifteen cases of dural defect were found intraoperatively,including 11 males and 4 females,aged from 40 to 68 years (average,55.6 years).Preoperative CT scans were earefully analyzed.During the operation,the ossified mass was excised or floated and the arachnoid was reserved in order to reduce dural damage.Dural defects were repaired by suturing or covering with muscle.After surgery,patients were confined to bed rest to allow for drainage or puncture.Results All 15 patients were followed up for 12 to 18months (average,14.8 months).After operation,10 patients recovered fully without CSFL.Five patients developed CSFL,including 4 cases of spinal dural mater injury that healed within 4-6 days with bed rest and pressure dressing,and 1 case of cerebrospinal fluid pseudocyst that disappeared within 3 weeks with repeat puncture and aspiration treatment.No cases required secondary operations or shunt placement.All cases exhibited good neurological improvement.Conclusion During anterior surgical treatment for severe cervical OPLL,CSFL can be effectively prevented through eareful analysis of preoperative CT images,meticulous operative technique,reasonable handling of the ossified mass and positive repair of dural damage.Moreover,postoperative bed rest and drainage are effective to treat CSFL.

Objective To summarize the characteristics and clinical value of multi-slice spiral computed tomography (MSCT) examination in the biliary gallbladder-duodenal fistula.Methods The imaging data of 28 patients with gallbladder-duodenal fistula who were admitted to the Wuxi No.2 Hospital of Nanjing Medical University between June 2012 and March 2015 were retrospectively analyzed.All the 28 patients received MSCT examinations,and the imaging changes were observed and analyzed,including the location of lesions,figures of fistulous tract,shrinking or enlarging gallbladder,pneumotosis and stones of gallbladder or bile duct.Results Of the 28 patients,fistula located at the duodenal bulb were detected in 14 patients,junction of the bulb and the descending part of the duodenum in 2 patients,ascending duodenum in 7 patients,horizontal part in 5 patients.Indirect signs of biliary gallbladder-duodenal fistula included that gallbladder volume in 28 patients was significantly reduced,cross sectional area of gallbladder was 2 cm × 1 cm-6 cm × 2 cm,and gallbladder wall was thickened with an average thickness of 5 mm (range,4-9 mm).Adhesion of gallbladder and duodenum,unclear boundary,structure disorder and visible effusion surrounding gallbladder were detected.Among 21 patients with biliary gas,19 patients had pneumotosis of gallbladder and 17 had biliary pneumatosis.Biliary stones were detected in 23 patients including cholecystolithiasis in 19 patients,gallbladder neck stones in 6 patients,common bile duct stones in 13 patients and intra-and extra-hepatic cholangiolithiasis in 1 patient.The diverticulum signs appeared in the duodenum of 11 patients.The direct signs of MSCT in the biliary gallbladder-duodenal fistula included that fistulous tract of 13 patients clearly showed and some were dumbbell-shaped.Two and 2 patients were complicated with gallstone ileus and multiple liver abscesses,respectively.The diagnostic results of MSCT in 28 patients were compared with the results of operative exploration,with an diagnostic concordance rate of 78.6％ (22/28),and the diagnostic concordance rate of gallbladder stones was 82.1％ (23/28).Conclusions The indirect signs of MSCT in patients with biliary gallbladder-duodenal fistula include pneumotosis of gallbladder or/ and biliary gas,gallbladder neck stones or common bile duct stones,gallbladder shrank,adhesion of gallbladder and duodenum,unclear boundary,diverticulum signs in the adhesions of duodenum and gallbladder,and clear orificium fistulae of gallbladder-duodenum is a direct sign of MSCT.