Objective To establish a bladder cancer MDR cell line by gene transfection and to lay a foundation for the research of occurrence and reversion mechanisms of MDR. Methods The invasive bladder cancer T24 cells were transfected with mdr1 cDNA by cationic liposome (DOTAP) introduced gene transfection.The adriamycin (ADM) resistant cells were screened as MDR cells,named TADM. The MDR phenotype of TADM was identified by MTT, immunohistochemistry,immunofluorescence,flow cytometry,PCR and RT PCR. Results The relative resistant index of TADM was 41.6,mdr1 cDNA being integrated into the genome of T24.As a result,the expression of P gp and mdr1 mRNA in TADM increased. Conclusions The integration of mdr1 cDNA into the T24 cells greatly increases the expression of P gp.TADM cells manifests excellent MDR phenotype.The establishment of MDR cell lines by gene tranfection has the benefits of less time consuming,stronger drug resistivity and more stable.

Objective To evaluate the efficacy and safety of interventional occlsu ion operait no by analyizn g thes urgical data of 23 csa es of infants with patent ductusa rteriosus complicated with middle and severe pulmo an ry arterial hypertension.Methods Teh 23 cases of infants wiht patne t ductus arteriosus com-plicated with middlea nd severe pulmonary arterial hypertensionw ere collected in the hospital from January 2011 to December2014 .These infatn s rce eived transcateh ter occlusion with intravenuo s anesthesia after the preoperative examination.The operation procse s included:right ventriculography and pulmonary atr ery pressure tested,then lateral angiogar phy of descending aorta was performed to observe the type and size of patent ud ctus atr eriosus and measure ascending aorta,descending aortic pressure,and recorded the pressure re-spectively.1 ml blood sample of ascending aorta,pulmonary artery and inef rior vena vein respectively was used for gas analysis.All these data was used to calculate pulmonary vascular resistance.After tried to plug-ging effectiveyl we can release the occluder.In the postoperative 24 h,1 month,3 months,the infants should be measured with Doppler echocardiography,chest X ray and electrocardiogram examination.Results The clinical symptoms disappeared and the short-term follow-up was not associated with the complications of interventional therapy.Th e comparison of the pressure changes before and after the operation were performed as following, aortic per ssure decreased [ preoperation ( 68.3 ±17.5 )/( 21.4 ±3.7 ) mmHg, postoperation (52.4 ±8.7)/(15.6 ±3.5) mmHg,1 mmHg=0.133 kPa],ascending aorta pressure increased(preoperation (83.5 ±5.9)/(51.3 ±3.6) mmHg,postoperation(88.2 ±5.1)/(52.4 ±2.7) mmHg),and descending aorta pressure increased ( preoperation ( 81.4 ±3.3 )/( 48.2 ±2.7 ) mmHg, postoperation ( 86.5 ±4.7 )/(51.5 ±3.2) mmHg), the differences were statistically significant before and after surgery ( t =5.455/3.945 ,P<0.01;t=-2.696/-1.193 , P<0.05; t=-4.167/-3.745 , P<0.01 ) .Conclusion Under conditions of mastering the appropriate operation time and strengthening the management of the perioperative management,transcatheter measurement is safe and effective for infants with patent ductus arteriosus compli-cated with middle and severe pulmonary arterial hypertension.

Objective To observe the change of intracellular rhodamine 123 distribution model in bladder cancer sensitive cells(T24) and resistant cells(TADM),and to explore the cause of the change and its relation with multidrug resistance(MDR). Methods T24 and TAMD cells were cultured together with rhodamine 123 for a certain period.The intracellular distribution and intensity of fluorescence were detected by interactive laser cytometer(ILC). Results In T24 cells,rhodamine 123 was located mainly in the perinuclear membrane area,and in TADM cells,rhodamine 123 was concentrated mainly at the two poles of nucleus.After the extracellular rhodamine 123 was washed,in T24 cells ,the intracellular fluorescent intensity decayed slowly.It took more than 40 minutes for T24 cells to eliminate intracellular rhodamine 123 completely.By contrast,in TADM cells, the intracellular fluorescent intensity decayed rapidly.It took only 15 minutes for TADM cells to eliminate intracellular rhodamine 123 completely. Conclusions (1)Intracellular rhodamine 123 was rapidly pumped out,which is the main cause of MDR in TADM;(2)The intracellular rhodamine 123 distribution model in TADM differ from that in T24,which may be another cause of its MDR phenotype.

Objective To study the clinical value of heat shock protein (HSP)70 in the diagnosis of neonatal asphyxia and the correlation of HSP70 and Neonatal Behavioral Neurological Assessment (NBNA)score.Methods From January 2014 to June 2016,full-term neonates born in our hospital were enrolled in the study and assigned into mild and severe asphyxia groups.Normally delivered full-term infants were assigned to the control group.Blood from umbilical artery were extracted immediately after birth and HSP70 levels were detected using ELISA.The NBNA scores were recorded at the 7th,14th and 28th-day after birth.Results HSP70levels in both mild (n =46 )and severe (n =35 )asphyxia groups were significantly higher than the control group(n =50)[(14.4 ±2.7)ng/ml、(17.7 ±4.5)ng/ml than(11 .9 ± 2.3)ng/ml,P <0.05].The severe asphyxia group had even higher HSP70 levels than the mild asphyxia group (P <0.05).The NBNA scores of both asphyxia groups were significantly lower than the control group (P <0.05).The umbilical pH values of both two asphyxia groups were also significantly lower than the control group(P <0.05).Correlation analysis showed that HSP70 level was negative correlated with NBNA score (7th,14th,28th-day)(r =-0.574、-0.493、-0.208,P <0.05).The HSP70 level was negatively correlated with umbilical pH (r =-0.576,P <0.05).The area under curve(AUC)for HSP70 levels to predict asphyxia was 0.798(95%CI 0.722 ~0.874,P <0.05).Conclusions HSP70 level in umbilical cord blood can be used as an indicator for neonatal asphyxia.The more severe the asphyxia,the higher the HSP70 levels and the lower NBNA score and umbilical pH.

Objective To screen the neonatal malrotation with PITX2 gene exon 2 and 5 gene mutation through the study on molecular genetics.Methods From January 201 2 to December 201 4,1 5 cases of neonatal malro-tation infants(experimental group)and 25 healthy newborn infants(healthy control group)were selected as the research subjects from the First Affiliated Hospital of Shihezi University Medical College.The experimental group included 1 5 ca-ses of volvulus,4 cases of volvulus with duodenal atresia and 3 cases of volvulus with jejunal atresia.The clinical fea-tures were recorded and 3 mL peripheral venous blood from each subject was collected.After ethylenediamine tetraacetic acid (EDTA)anticoagulation,genomic DNA was extracted.Polymerase chain reaction (PCR)was used to amplify the exon 2 and exon 5 of PITX2 gene,and the direct sequencing method was used to screen whether there were mutations in these 2 loci.Results According to the findings of the matching gene,PITX2 gene exon 2 and exon 5 mutations were not detected in 15 cases with intestinal malrotation of the experimental group and 25 healthy newborns in the healthy control group.Conclusions Polymorphisms is not detected in PITX2 gene exon 2 and exon 5 in small groups of newborn,but this does not exclude the possibility the gene caused newborns suffering from intestinal malrotation by other means.

Objective To characterize the dimerization and the antigenicity of the ORF2 polypep-tide of hepatitis E virus (HEV, genotype 4). Methods HEV ORF2 gene was cloned from the serum of a patient with hepatitis E. The genotype was determined by sequencing. Three ORF2 polypeptides differing in size and other polypeptides with point mutations were produced in E. coli. The recombinant polypeptides were purified and analyzed by SDS-PAGE and Western blot. Results The ORF2 polypeptide containing 459-607 amino acid formed homedimer even in 8 mol/L urea. The truncated polypeptides containing amino acid 472-607 or 459-594 formed monomer only. The mutations at amino acid 562 or 595 disrupted the ho-modimer, whereas the mutations at amino acid 476 or 580 did not. Anti-HEV from hepatitis E patients only reacted with the homodimer form of the polypeptide 459-607 and did not react with monomer or tnmcated pol-ypeptides. Conclusion The amino acid 459-607 of HEV ORF2 is essential for dimerization of the ORF2 polypeptide. Residues at amino acid 562 and 595 are critical for the dimerization. The antigenicity of the polypeptide 459-607 mainly depends on its homodimer form.

Objective To compare the dosimetric difference among plans designed by 4-field,6-field TomoDirect and TomoHelical techniques in Tomotherapy system for left-breast cancer patients with radiotherapy after breast-conserving surgery.Method A total of 16 patients with left-breast cancer following breast-conserving surgery and intensity-modulated radiation therapy were enrolled in this retrospective study.The 4-field TomoDirect (TD4),6-field TomoDirect (TD6),and TomoHelical (TH) techniques were applied to design simulation plans in tomotherapy system for each patient,respectively.The differences of dose distribution and treatment parameters were analyzed in this study.Results Three plans all met the clinical requirement.Thereinto,TD4 was superior to TH in the dose limitation of organs at risk (OARs),especially the max dose of cord and right-breast,thc 5 Gy radiation volume of lung,and the mean dose of heart(F =595.60,129.24,60.44,65.37,P ＜ 0.05),but inferior to TH in dose homogeneity (HI) and conformity (CI) (F =2.78,60.93,P ＜ 0.05).However,TD6 improved TD4's HI and CI when delivered the lower OARs dose compared to TH.Meanwhile,the number of monitor units was less in TD technique and reduced the treatment times (F =24.89,3.75,P ＜ O.05).Conclusions For the radiotherapy of left-breast cancer patients after breast-conserving surgery,TD6 technique appeared to be superior,with the lower radiation dose of OARs compared to TH technique,and the better target's HI and CI in comparison with TD4 technique,especially in patients with early stage breast cancer.

Aim To determine the effective compo-nents of Semen Ziziphi Spinosae for sedative-hypnotic and its mechanism. Methods The extraction of Se-men Ziziphi Spinosae and the rat brain homogenates were prepared. High concentrations of Diazepam com-petitively replaced the ligand compounds of Semen Ziz-iphi Spinosae combining BDZ receptor in brain tissue, and all the compounds with sedative and hypnotic effects were collected and identified by HPLC and LC-MS technique, as the compounds extracted from the brain tissue were administered with Semen Ziziphi Spi-nosae. The brain tissue was administered with Diaze-pam, and with Semen Ziziphi Spinosae and Diazepam. Results The HPLC chromatograms show that the peak time of BDZ receptor ligand compounds was 2. 71 min and 46. 87min, when compared with Diazepam. And the LC-MS chromatograms display the relative molecu-lar weight of the ligand compounds was 274. 28 m/z, 453. 34 m/z,496. 34 m/z and 608. 38 m/z respective-ly. According to the fingerprint of Semen Ziziphi Spi-nosae, these compounds may be fatty acid substances and lupine pill triterpene compounds. Conclusions On the basis of the principle of receptor ligand bind-ing, we established a way to quickly analyze and iden-tify the role of natural products in the same drug target compounds. The method not only can clearly define the effective components of natural products, but also clar-ify the mechanism of action of the compounds. The ac-tive ingredient of calm hypnosis in Semen Ziziphi Spi-nosae may be fatty acid substances Palmitic acid ( C16 H32 O2 ) and lupine pill triterpene compounds Alphitolic acid( C30 H48 O4 ) and Spinosin( C28 H32 O15 ) . They exert their sedative and hypnotic effects by combining with BDZ receptor, and the research has laid a theoretical foundation for the further study about mechanism of Se-men Ziziphi Spinosae.

Objective This study observed the clinical curative effects of aldose reductase inhibitor epalrestat in treat-ment of diabetic sensory neuropathy. Methods Thirty-four diabetic sensory neuropathy patients were selected. The nerve electrophysiological data were collected before and after treating with epalrestat. Results The ratios of nerve con-ductive velocity in peroneal nerve sensory showed slowing down than normal before and after the treatment, which were respectively 61% and 32%; The ratios of the nerve conductive velocity in tibial nerve sensory nerve segment 1 showed slowing down before and after the treatment,which were respectively 97% and 65%; The nerve conductive velocity of the peroneal sensory nerve after treatment was significantly faster than that before treatment, the nerve conductive ve-locity of the the tibial nerve sensory nerve motion segments in 1 after treatment was significantly faster than that before treatment. Conclusion Epalrestat is one of the effective methods in the treatment of diabetic peripheral neuropathy.