Objective:To analyze the curative effect of suppression expression of URG4 on cervical cancer mice and the possible mechanisms.Methods:The expression of URG4 were analyzed in cervical cancer tissues and para-carcinoma tissues.16 male BALB/c nude mice were divided into Group A ( inoculated with URG4-siRNA SiHa cells) and Group B ( inoculated with SiHa cells) . After 4 weeks,mice were executed and analyzed.Results:The expression of URG4 were higher in human cervical cancer tissues than that in para-carcinoma tissues ( P<0.05).After experiment,Group A had a lower tumor weight,but a higher body weight than these of Group B,the difference was statistically significant (P<0.05).Group A mice had a lower levels of URG4 in tumor than that of Group B mice (P<0.05).Group A mice had a lower levels of Ki67 in tumor than that of Group B mice,the difference was statistically significant (P<0.05).Group A mice had lower levels of CyclinD1,β-catenin and C-myc in tumor than these of Group B mice (P<0.05). Conclusion:The expression of URG4 in cervical cancer tissue is up-regulation.Suppression expression of URG4 can suppress tumor growth in vivo,which may though suppress the Wnt/β-catenin signaling pathway.

Objective To analyze the clinical effect of minimally invasive percutaneous nephrolithotomy titanium laser lithotripsy(MPCNL)on upper ureteral calculi.Methods According to digital table,68 patients with upper ureteral calculi were randomly divided into the study group and control group,cach group in 34 cases.The study group received the MPCNL treatment,the control group received extracorporeal shock wave lithotripsy(ESWL)therapy. The therapeutic effects were compared.Results The success rate of gravel,stone clearance rates of the study group were 91 .1 8% and 91 .1 8%,which were higher than 70.59%,67.65% of the control group,there were significant differences between the two groups(χ2 =4.660,5.757,P =0.031 ,0.0164).Conclusion Percutaneous nerve titanium laser lithotripsy for upper ureteral stones can achieve good therapeutic effect.

Objective To investigate the clinical features,critical laboratory parameters,and fetal monitoring methods in intrahepatic cholestasis of pregnancy(ICP).Methods A retrospective analysis of 21 cases of ICP suffered with fetal death in Women's hospital.School of Medicine.Zhejiang University from January 1999 to December 2010 were discussed.Results(1)The average age of ICP patients suffered with fetal death were(30.2±4.6)years old.Among them,4 cases were older than 35 years,six cases were multipara.oneo of them suffered stillbirth 2 year before.Twenty cases were singleton pregnancies and 1 cage was twin pregnancy.(2)All 21 cases of fetal death occurred in the third trimester,12 cases occurred before 37 weeks,9 cases after 37 weeks.Nine cases were diagnosed by ultrasound in outpatient clinics,fetal heart beat disappeared in 9 patients after admission because of ICP, two disappeared after labor, one during anesthesia before emergent surgery. Perinatal mortality rate of ICP was 0. 148% (21/14 184), and fetal death occurred from 29 to 41 weeks with an average gestational age of ( 33.8 ± 4. 2 ) weeks, ( 3 ) Puritus occurred in all 21 cases while 11 of them had pruritus all over the body. Ten pregnant women felt the fetal movement decreased or disappeared before diagnosis of fetal death. The glycocholic acid levels increased in all of the 21 cases. Among them, glycocholic acid levels in 11 cases were (21.49 -64. 48) μmol/L, while in 10 cases were ≥64. 48 μmol/L Serum bile acid levels elevated in 16 cases which had been analyzed ( the other 5 cases had not been checked ), and the highest level reached 270 μmol/L Serum alanine aminotransferase and aspartate aminotransferase were increased in 14 cases. Seven cases had their total bilirubin ＞21 μmoL/L, and 12 cases had their direct bilirubin levels significantly elevated. Among the 21 cases of ICP, 15 cases were in severe status, while the other 6 cases were mild. (4) Nine patients had no antepartum surveillance since fetal death were diagnosed before admission. The results of antepartum surveillance were as follows: 2 cases had nonreassuring nonstress test (NST), one had mild "V" type deceleration. Absence of diastolic flow in umbilical artery were found in 3 cases, and low fetal biophysical score was got in one case. ( 5 ) All 21 patients had vaginal delivery. Six of them delivered after natural contraction, and the remaining 14 cases delivered after oral intake of mifepristone and amniotic injection of ethacridine, or oxytocin induced labor within 48 hours, only one case delivered after additional dinoprostone suppositories. The appearance of fetus, placentas and membranes were normal, the lengths of umbilical cord were average. Four cases were found with cords binding the necks or the bodies. Eighteen cases had grade Ⅲ amniotic fluid with meconium-stained, and 2 cases complicated by oligohydramnios. Ten cases had their fetuses and placentas examined by pathologist. Among them, one case had multiple malformations, no more obvious pathological abnormalities were found in other fetuses. Pathologic examination showed that fibrin deposited around chorion and deciduas basalis, large vessels accompanied by calcification, degeneration,hemorrhagic infarction, and increased focal syncytial nodules could be seen in all of the ten placentas. Conclusions Fetal death in pregnant women with ICP of ten occurs after the contractions, Severe ICP may be a key factor that involved in the occurrence of fetal death. Up to now, there is no valid indicators in fetal monitoring, which can predict fetal death. Extensive assessment of the severity and careful antepartum surveillance should be achieved before timely termination of pregnancy.

Objective To optimize in vitro amplification of human γδ T cells with cytokines for tumor adoptive immunotherapy.Methods On the basis of the immobilized anti-TCR γδ antibody plus IL-2 system, other γ chain receptor family cytokines, including IL-7, IL-15 and IL-21, were tested to amplify human peripheral blood γδ T cells either alone or in diversity combination.The percentage of γδ T cells was measured by flow cytometry, and the proliferation efficiency of γδ T cells was calculated.The expression of proliferation-or cytotoxicity-related molecules on γδ T cells was examined by flow cytometry in order to explore the relevant mechanisms.The cytotoxicity of γδ T cells to Daudi cells was detected by lactate dehydrogenase.Results IL-15 alone but not IL-7 or IL-21 increases the γδ T cell purity, amplification efficiency and cytotoxicity to reach comparable levels to those of IL-2.IL-2 plus IL-15 up-regulates the expression of CD69 on γδ T cells and significantly increases their amplificationefficiency (P<0.05).IL-2 plus IL-21 enhanced the cytotoxicity of γδ T cells against Daudi cells by increasing the expression of granzyme A (P<0.001).The combination of IL-2, IL-15 and IL-21 significantly improves cytotoxicity of γδ T cells but reduces their amplification efficiency.In addition, when IL-21 was applied for a short time, it also enhanced the cytotoxicity of γδ T cells (P<0.05).Conclusions The combination of IL-2 and IL-15 as well as a short time addition of IL-21 is the best cytokine recipe to amplify human peripheral blood γδ T cells in vitro with immobilized anti-TCR γδ antibody, which can increase both the proliferation efficiency and the cytotoxicity to tumor cells of γδ T cells.

Objective To investigate the relationship between the serum homocysteine(Hcy) level and heart rate variability in elderly patients with metabolic syndrome.Methods Retrospective analysis of 162 elderly patients with metabolic syndrome aged 60 and above was conducted in our hospital from January 2013 to August 2016.And the level of serum homocysteine and other biochemical indicators were measured.The 162 cases were divided into three groups of 42 cases in Hcy normal group(Hcy,5-15 μmol/L),70 cases in Hcy mildly high group(Hcy,16-30 μmol/L),50 cases in Hcy moderately high group (Hcy,31 ～ 100 μmol/L).Simultaneously 24-hour rhythm in ambulatory heart rate variability spectrum was used to detect HRV.And the relationship between heart rate variability(HRV) and Hcy levels were analyzed.Results Normal RR interval standand deviation (SDNN),5 min mean heart rate standard deviation (SDANN),Edinburg index (PNN50),root mean square value of adjacent RR interval difference(RMMSD)were all lower in Hcy mildly high group and Hcy moderately high group than in the Hcy normal group (P＜0.05).Low-frequency(LF) and highfrequency(HF)were lower in Hcy mildly high group and Hcy moderately high group than in the Hcy normal group(P ＜ 0.05).Sympathetic-vagal balance indicators (LF/HF) were higher in Hcy mildly high group and Hcy moderately high group than in the Hcy normal group (P＜ 0.05).The HRV indexes were lower in Hcy high groups than in the Hcy normal group.Hcy was negatively correlated with SDANN,RMSSD,LF,HF,and LF/HF at night(r=-0.578、-0.651、-0.531、-0.662、-0.729,all P ＜ 0.01).Conclusions The elevated homocysteine level and metabolic syndromecombined with decreased HRV in the elderly patients suggest that cardiac autonomic nervous system might be injuried.Enhancing the detection of HRV and homocysteine level in elderly patients with MS is clinically useful for early detection of autonomic neuropathy.

In order to study the expression and the feasibility of scaled production of neuropeptide in the routine expression system such as E.coli with the pituitary adenylate cyclase activating polypep tide(PACAP) as an example, the following experiments were carried out. First, on the basis of the reported amino acid sequence of PACAP, DNA sequence of PACAP w as deduced and six partially complementary oligonucleotide fragments were design ed. The coding region of PACAP was obtained by renaturing the DNA fragments and ligation and identified by DNA sequencing. The coding region of PACAP was cloned into plasmid pGEX-4T-3 and transformed into E.coli BL21(DE3 ). An expression strain BLPACAP was selected. SDS-PAGE analysis revealed that t he GST-PACAP fusion protein was highly expressed and accumulated to about 30% o f the total bacterial proteins. By affinity chromatography, up to 90% GST-PACAP was purified by one step from bacterial lysate. The purified protein could prom ote neurite outgrowth of PC12 cells and the survival of spinal cord neurons.

Objective To investigate the modulating effects and explore their mechanism of 9-nitrocamptothecin and its liposomes to induce apoptosis and inhibit cell cycle in HepG2 and L02 cell lines. Methods Cells were incubated with 9-nitrocamptothecin(9NC) or with 9-nitrocamptothecin liposomes for 24 h, 48 h and 72 h, then, the cell viability was measured via MTT assay; cell cycle and apoptosis was evaluated by flow cytometry after stained by PI and Annexin V-PE/7AAD. Additional, Western Blot was used to evaluate the expression of cell cycle and apoptosis related protein. Results Both cells viability were apparently inhibited by the 9-nitrocamptothecin and 9-nitrocamptothecin liposomes, the inhibitory effect showed a time-dependent and dose-dependent manner. Both S and G2/M phases arrest were observed after incubated with drugs. HepG2 cell was completely arrested in S phase when 9NC concentration over than 0. 1 μmol/L after incubation for 24 h, while more than 95% cells arrested in G2/M phase when 9NC concentration is 0. 1 μmol/L after incubation for 72 h. Apoptosis induction effect also showed a time-dependent and dose-dependent manner. Western Blot results showed the expression of Bax and Caspase-3 were upregulated while Cyclin A, Cdk2, Cyclin E and Bcl-2 were downregulated. More importantly, the compounds were more cytotoxic to the cancer cell lines than to the normal liver cell. Conclusions 9-nitrocamptothecin and 9-nitrocamptothecin liposomes can potently inhibit cell growth via regulation of cell cycle and induction of apoptosis, and this effect was preferentially in cancer cell. Inhibitory of 9-nitrocamptothecin liposomes was slightly better than the 9-nitrocamptothecin.

Objective To observe the inhibitory effect and mechanism of 9-nitrocamptothecin liposomes on HepG2 liver carcinoma cells. Methods HepG2 cells were incubated with 9-nitrocampto-thecin(9NC) or with 9-nitrocamptothecin liposomes(9NC-LP) for 24 h, 48 h and 72 h. Cell viability was then measured by the MTT assay. Cell cycle and apoptosis were evaluated by flow cytometry.Western Blot was used to determine the expression of cell cycle and apoptosis related proteins. HepG2tumor-bearing mouse models were then established. The HepG2 tumor-bearing mice were randomly divided into control group, free liposomes group, DMSO group, 9NC low dose group, 9NC high dose group, 9NC-LP low dose group and 9NC-LP high dose group. There were 10 mice in each group.Drugs were administered by tail vein and tumor volume and body weight were observed 28 days after administration. Then animals were sacrificed and the expression of proteins from tumor homogenates was analyzed by Western blotting. Results In vitro, HepG2 cell viability was apparently inhibited by 9NC and 9NC-LP, and the inhibitory effect increased in a time-dependent and dose-dependent manner.Both S and G2/M phase arrests were observed after incubation with drugs. HepG2 cells were completely arrested in S phase with 9NC concentration over than 0.1 μmol/L after incubation for 24 h,while more than 95% of cells arrested in G2/M phase when 9NC concentration was 0.1 μmol/L after incubation for 72 h. In vivo, compared with the control group, the average tumor volume was reduced in both the 9NC and 9NC-LP group (P<0.05) , and the average animal body weight also decreased in both the 9NC and 9NC-LP group (P<0.05). There was no significant difference among the control group, free liposomes group, and DMSO group. The lights inhibition rates of tumor growth in the 9NC-LP(2.5 mg/kg),9NC-LP(1.5 mg/kg),and 9NC(1.5 mg/kg)groups were 87.02%, 51.57%and 35.47%, respectively. In the 9NC-LP(2.5 mg/kg)group, >50% of animals died 14 days after drug administration. Conclusion 9NC and 9NC-LP can inhibit HepG2 cell growth via cell cycle arrest and apoptosis induction. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo,which means 9NC-LP is a promising compound for cancer therapy via intravenous administration.

Objective To analysis the relationship between gestational age and perinatal outcomes in patients complicated with early onset severe preeclampsia.Methods Retrospective study was conducted on clinical documents of 221 patients with early onset severe preeclampsia( ＜ 34 weeks) who delivered after 28 gestational weeks in Peking University First Hospital from July 1999 to June 2009.Patients were divided into three groups based on gestational weeks at delivery: group Ⅰ (n = 81 ) delivered at 28 -31 weeks+6,group Ⅱ (n = 78) at 32 -33 weeks+6 and group Ⅲ (n = 62) after 34 weeks.The clinical characteristics and perinatal outcomes were compared among those three groups.Results ( 1 ) Outcome of neonates:Among 221 neonates, 13 neonates lost follow-up, including 9 in group Ⅰ , 3 in group Ⅱ, 1 in group Ⅲ.The incidence of neonatal respiratory distress syndrome ( RDS ) of 26% ( 19/72 ) in group Ⅰ were significantly higher than 7% (5/75) in group Ⅱ and 10% (6/61) in group Ⅲ (P ＜ 0.05 ).The neonatal mortality rate of (43% ,31/72) in group Ⅰ were significantly higher than 3% (2/61) in group Ⅲ and 28%(21/75) in group Ⅱ (P ＜0.05 ).The incidence of maternal complications showed no statistical difference among three groups.(2) Neonatal death analysis: all neonatal death were due to parents' give up, including 26%(8/31) in group Ⅰ, 67% (14/21)in group Ⅱ and 1/2 in group Ⅲ, which reached statistical difference(P＜0.05).Conclusions The incidence of neonatal RDS in mother with early onset severe preeclampsia was decreased if delivered after 32 weeks, and the perinatal mortality was remarkably decreased if delivered after 34 weeks.Therefore, the perinatal survival rate in women with early onset severe preeclampsia can be improved by minimizing the impact of social factors.

Objective To explore the influence of different positive end-expiratory pressure (PEEP) levels on cerebral blood flow (CBF) and cerebrovascular autoregulation in patients with acute respiratory distress syndrome (ARDS).Methods A prospective study was conducted.Moderate or severe ARDS patients admitted to Department of Critical Care Medicine of Jiangxi Provincial People's Hospital from January 1st,2013 to October 1st,2013 were enrolled.The changes in hemodynamics,respiratory mechanics and gas exchange under different levels of PEEP were observed.CBF velocity of middle cerebral artery (MCA) was measured using transcranial Doppler (TCD),and breath-holding index (BHI) was also calculated.Results 35 patients with ARDS were included.The oxygenation index (OI),peak inspiratory pressure (PIP),plat pressure (Pplat) and central venous pressure (CVP) were markedly elevated [OI (mmHg,1 mmHg=0.133 kPa):324.7± 117.2 vs.173.4± 95.8,t=5.913,P=0.000; PIP (cmH2O):34.7 ± 9.1 vs.26.1 ± 7.9,t=4.222,P=0.000; Pplat (cmH2O):30.5 ± 8.4 vs.22.2 ± 7.1,t=4.465,P=0.000; CVP (mmHg):12.1 ± 3.5 vs.8.8 ± 2.2,t=4.723,P=0.000] when PEEP was increased from (6.4 ± 1.0) cmH2O to (14.5-± 2.0) cmH2O (1 cmH2O=0.098 kPa).But no significant difference in the heart rate (beats/min:85.5 ± 19.1 vs.82.7 ± 17.3,t=0.643,P=0.523),mean arterial pressure (mmHg:73.5 ± 12.4 vs.76.4 ± 15.1,t=0.878,P=0.383) and CBF velocity of MCA [peak systohc flow velocity (Vmax,cm/s):91.26 ± 17.57 vs.96.64 ± 18.71,t=1.240,P=0.219; diastolic flow velocity (Vmin,cm/s) 31.54 ±7.71 vs.33.87 ±8.53,t=1.199,P=0.235; mean velocity (Vmean,cm/s) 51.19 ± 12.05 vs.54.27 ± 13.36,t=1.013,P=0.315] was found.18 patients with BHI＜0.1 at baseline demonstrated that cerebral vasomotor reactivity was poor.BHI was slightly decreased with increase in PEEP (0.78 ± 0.16 vs.0.86 ± 0.19,t=1.905,P=0.061).Conclusions Some of moderate or severe ARDS patients without central nervous system disease have independent of preexisting cerebral autoregulation impairment.However,independent of preexisting cerebral autoregulation may not further be impaired when a high PEEP was chosen.