Objective:This study aims to evaluate the effect of an elevated preoperative neutrophil to lymphocyte ratio (NLR) on outcome after comprehensive staging laparotomy or optimal tumor debulking surgery for epithelial ovarian cancer (EOC) and determine the value of the NLR as an independent prognostic prediction marker. Methods:A total of 80 women with primary EOC and with complete clinical and pathological information documented at the time of surgery were selected for this study. The optimum cut-off value of the preoperative NLR was identified through receiver operator characteristic (ROC) curve, and the patients were then classified into two groups: low and high NLR group. Univariate and multivariate analyses were performed to assess the prognostic effect of the preoperative NLR patients who underwent comprehensive staging laparotomy or optimal tumor debulking surgery. The levels of expression of CD68 were measured through immunohistochemistry. Results:The optimal cut-off value of the NLR was 3.8. The preoperative NLR differed significantly in the FIGO stage between the low NLR group (NLR ≤3.8) and the high NLR group (NLR>3.8), but no discrimination was observed in other parameters. The mean follow-up time was 45 months, and the post-operative 1-and 3-year survival rates were 93.7%and 60.0%, respectively. The preoperative NLR>3.8 and stageⅢ/Ⅳwere all risk factors for poor overall and disease-free survival. Multivariate analysis revealed the patients with high NLR (P<0.05) and stage Ⅲ/Ⅳ (P<0.05) had prognostic significance for poor overall survival. The number of CD68-positive tumor-associated macrophages was significantly higher in the high NLR group than in the low NLR group (54.65 ± 8.78 and 41.78 ± 9.10, respectively; P<0.001). Conclusion: An elevated blood preoperative NLR indicates poor prognosis in patients with EOC. Preoperative NLR may function as an important independent prognostic factor for patients with EOC.

Objective To explore the effect of ginseng combined with transplantation of bone marrow mesenchymal stem cells(BMSCs)on functional recovery of rats with spinal cord injury(SCI). Methods Forty-eight female Wistar rats were randomly assigned to following four groups(n=12 per group):the sham operation group was treated by opening the vertebral lamina and exposing spinal cord without SCI;the SCI model group was reproduced by using improved Allen bump method and afterwards no treatment was given;methyl prednisolone(MP)group was treated by MP pulse treatment after SCI,including intravenous injection of MP 30 mg/kg immediately after SCI and 4 hours later the same injection was repeated,and then the same intravenous injection 2 times daily,3 days in total;The ginseng+BMSCs group was treated by orally taking ginseng ultra-microgranules 300 mg/kg after SCI,twice a day for 20 days and BMSCs 5μL(concentration 1×107 cell/μL)transplantation was carried out in SCI region on the 7th day after SCI. In the above 4 groups,the ethological observation(BBB scores)was done regularly and on the 30th day after operation,silver staining was applied to investigate the changes of spinal cord,and neuro-electrophysiological tests including somatosensory evoked potential(SEP)and motor evoked potential(MEP)were performed. Results In sham operation group,after surgery the movement of both hind limbs became temporarily sluggish and on the 7th day their functions recovered to approximately normal. In SCI model group,after injury paralysis of both hind limbs occurred,while in the MP group and ginseng+BMSCs group,different degrees of functional recovery of the injured limbs developed,and the recovery in ginseng + BMSCs group was more significant. Compared with sham operation group,after surgery the BBB score was reduced markedly at various time points in SCI model group;compared to the SCI model group,the BBB scores in MP and ginseng+BMSCs groups were increased significantly,especially more remarkable in ginseng+BMSCs group(all P<0.05),and beginning from the 12th day after operation,the difference became obvious (5.23±1.22 vs. 3.61±1.03, P<0.05). Histological detection showed that in sham operation group,the structure of spinal cord was complete,neurons distributed evenly in the gray matter and a large number of silver staining positive nerve fibers paralleled to each other and arranged regularly;in SCI model group,fragmented construction was present and the defects of gray and white matters were prominent. Compared with the SCI model group, the extents of tissue necrosis in MP and ginseng + BMSCs groups were ameliorated. The neuro-electrophysiological tests demonstrated that in SCI model group,the loss of normal wave form occurred. Compared with SCI model group,in MP and ginseng+BMSCs groups,after treatment latent periods(ms)were shortened prominently in different degrees of SEP and MEP,and their peak-to-peak values(mV)were increased obviously;the improvement in potential in ginseng+BMSCs group was greater than that in MP group〔SEP:latent period(ms):3.31±0.36 vs. 4.66±0.33, peak-to-peak value(mV):0.10±0.01 vs. 0.05±0.01,MEP:latent period(ms):3.40±0.13 vs. 4.24±0.31, peak-to-peak value(mV):41.12±0.56 vs. 16.46±2.83,all P<0.05〕. Conclusion A combined treatment of ginseng and BMSCs transplantation can effectively promote the recovery of neural function for rats with SCI.

Objectives To determine the significance of serum total bile acid (TBA) in the diagnosis and treatment of infantile hepatitis syndrome. Methods Serum TBA and other liver function related items were determined with automatic enzymatic assay techniques in 67 with infantile hepatitis syndrome patients (age 26 days～7 months) and 100 normal infants (age 20 days～1 year). Results In the control group, the serum TAB level was 0～11.3 ?mol/L. TBA level was abnormal in 89.5% patients and the values were 0.5～226.0 ?mol/L ?s =(79 5?54.3) ?mol/L]. The difference was significant between the two groups. The TBA levels were well related to those of ALP, DBIL and ? GT and TBA was better than others in sensitivity and specificity. . The difference was significant between the two groups. The TBA levels were well related to those of ALP, DBIL and ? GT and TBA was better than others in sensitivity and specificity. Conclusion Serum TBA level is important in evaluating the diagnosis, treatment, and prognosis of infantile hepatitis syndrome.

Objective To investigate the clinical application value of Quadrant system combined with percutaneous pedicle screw fixation for treatment of lumbar degenerative disease.Methods The clinical data of 40 patients who sufferred from lumbar degenerative disease,were randomly divided into two groups:Quadrant system combined with percutaneous pedicle screw fixation to do discectomy and intervertebral bone graft fusion as invasive group (n =20),traditional posterior lumbar interbody fusion as conventional group (n =20).The operation time,length of incision,blood loss,postoperative drainage,JOA score of preoperation and postoperation,and effect (reforming Macnab standard) between two groups were recorded and compared.Results Operations were all well done in two groups.The operation time in invasive group was longer than that in traditional group [(147.3 ± 8.9) min,(136.7 ± 10.2) min,t =3.5,P ＜ 0.05].The length of incision [(3.65 ± 0.68) cm,(1 1.11 ± 1.29) cm,t =22.88,P ＜ 0.05)],blood loss [(205.00 ± 63.04) mL,(270.50 ± 77.58) mL,t =2.93,P ＜ 0.05],postoperative drainage [(90.3 ± 10.8) mL,(180.6 ± 1 3.9) mL,t =22.96,P ＜ 0.05] and days in hospital [(16.9 ± 2.0) days,(18.9 ± 2.1)days,t =3.05,P ＜ 0.05] in invasive group were all less.than those in traditional group (P ＜ 0.05).All cases had been followed up for 6-18 months,average of 13 months.There were no significant differences in excellent and good rate (reforming Macnab standard) between two groups in the last following up (P ＞ 0.05).There were no significant differences of JOA scores in preoperation [(12.0 ± 1.7) points,(11.0 ± 1.5) points,P ＞ 0.05],postoperative half a year [(22.0 ± 2.3) points,(21.0 ± 2.5) points,P ＞ 0.05],and postoperative one year [(23.0 ± 1.9) points,(22.0 ± 2.0) points,P ＞ 0.05].Conclusion Quadrant system combined withpercutaneous pedicle screw fixation is a safe,effective and minimally invasive surgical technique in treating lumbar degenerative disease.Compared with conventional group,invasive group has advantages such as less injury,less blood loss,simple operation,and good curative effect,which should be popularized.

Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase in bupivacaine-induced injury to neurons.Methods SH-SY5Y cells were seeded in 96-well culture plates at a density of 5× 104 cells/well and randomly divided into 3 groups (n=24each) using a random number table:control group (group C),bupivacaine group (group B),and apocynin (NADPH oxidase inhibitor) + bupivacaine group (group A+B).The cells were cultured in a serum-free medium in group C.The cells were cultured in a serum-free medium containing 1 mmol/L bupivacaine in group B.In group A + B,the cells were cultured for 30 min in a serum-free medium containing apocynin 100 μmol/L,and then cultured in a serum-free medium containing 1 mmol/L bupivacaine.At 2,4 and 6 h of incubation,the cells in 6 wells of each group were selected to evaluate the cell viability by MTS assay.At 4 h of incubation,the cells in 6 wells of each group were selected to detect reactive oxygen species (ROS) level by flow cytometry.Results Compared with group C,the cell viability was significantly decreased at 4 and 6 h of incubation,and the production of ROS was increased at 4 h of incubation in group B (P＜ 0.05).Compared with group B,the cell viability was significantly increased at 4 and 6 h of incubation,and the production of ROS was decreased at 4 h of incubation in group B (P＜0.05).Conclusion NADPH oxidase is involved in bupivacaine-induced injury to neurons.

Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.

Objective:The objective of this research is to study the serum level of the high-mobility group protein B1 (HMGB1) in human ovarian tumor (OvCa) and in a healthy control. This study also aims to identify different HMGB1 levels before and after sur-gery and to explore the inhibitory effect of HMGB1 gene silencing in the proliferation and invasion ability of OvCa. Methods: En-zyme-linked immunosorbent assay was used to measure the serum level of HMGB1 in OvCa patients and healthy subjects. Lentivirus vector with HMGB1 shRNA was constructed and used to infect OvCa cells. The expressions of HMGB1 mRNA and protein were test-ed by real-time PCR and Western blot. Cell proliferation was detected using the Cell Counting Kit-8 assay, whereas cell invasion and migration were detected by Transwell assay. Results:The serum level of HMGB1 was more elevated in patients with malignant diseas-es compared with individuals with benign diseases and the control groups. In the malignant group, the serum level of HMGB1 de-creased noticeably after therapy. Down-regulation of HMGB1 expression resulted in the inhibition of the biological behavior and metas-tasis of ovarian cancer cells. Conclusion: HMGB1 is closely associated with clinicopathologic features of OvCa. Knockdown of HMGB1 expression can significantly inhibit cell proliferation, cell migration, and cell invasion of OvCa. These findings indicate that HMGB1 can function as a therapeutic target for ovarian neoplasm in the future.

Objective To evaluate the effect of RNA interference in p53 gene on the expressions of genes involved in ultraviolet B (UVB)-induced premature senescence and photocarcinogenesis in human skin fibroblasts (HSFs).Methods A previously established HSF cell clone with repressed expression of p53,which was named as HSF-p53,was cultured and irradiated with a subcytotoxic dose (10 mJ/cm2) of UVB once a day for five consecutive days.The HSFs with normal expression of p53 served as the control.Subsequently,β-galactosidase (SA-β-gal)-staining was performed to estimate the degree of senescence,quantitative real-time PCR array was performed to determine the mRNA expressions of photocarcinogenesis-and senescence-associated genes,including p53,p21,p19,p16,pRb,fibronectin,osteonectin,smooth muscle 22 (SM22),bax,bcl-2,hypoxia-inducible factor-1 α (HIF-1α),vascular endothelial growth factor(VEGF),and human double minute-2 (hdm2).Statistical analysis was carried out by Student's t test using the software SPSS 10.0.Results The percentage of SA-β-gal-positive cells in irradiated HSF-p53 was 19.70％ ± 0.85％,significantly higher than that in unirradiated HSF-p53 (12.77％ ± 0.81％,t =6.45,P ＜ 0.05),but lower than that in irradiated control HSFs (50.48％ ± 5.30％,t =7.86,P ＜ 0.05),and similar to that in unirradiated control HSFs (18.50％ ± 0.45％,t =2.57,P ＞ 0.05).Compared with the control HSFs,the HSF-p53 showed decreased expressions of p21,p19,fibronectin,osteonectin,SM22 and bax genes (all P ＜ 0.05),but increased expressions of bcl-2,HIF-1α,VEGF and hdm2 genes (all P ＜ 0.05),and a similar expression of p16 gene (P ＞ 0.05); the repeated UVB radiation significantly promoted the expressions of p16 and pRb genes (both P ＜ 0.05),but had no obvious effect on the expressions of the other genes in HSF-p53 compared with unirradiated HSF-p53 (all P ＞ 0.05).Conclusions The inhibition of p53 expression may decelerate the UVB-induced premature senescence in HSFs,which may be involved in the p53-dependent tumor suppression.

Objective To evaluate the effect of methylprednisolone on hepatic ischemia-reperfusion (I/R) injury in the patients undergoing hepatolobectomy.Methods Sixty ASA physical status Ⅱ or Ⅲ patients,aged 30-64 yr,weighing 45-75 kg,scheduled for elective hepatolobectomy,were randomized to control group or methylprednisolone group (n =30 each).After induction of anesthesia,methylprednisolone 500 mg (in 100 ml of normal saline) was infused intravenously at 5 ml/min before skin incision in group M.Anesthesia was induced with propofol,fentanyl and cisatracurium.The patients were endotracheally intubated and mechanically ventilated.PETCO2 was maintained at 35-45 mmHg.Anesthesia was maintained with 1％-3％ sevoflurane inhalation,remifentanil infusion,and intermittent iv boluses of fentanyl and cisatracurium.MAP was maintained at 70-100 mmHg and HR at 50-90 bpm.At 10 min before induction of anesthesia,and on postoperative day 1,3 and 5,venous blood samples were collected for determination of the plasma levels of alanine aminotransferase (ALT),aspartate amminotransferase (AST),total bilirubin (TBIL),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6).Results Compared with group C,the plasma levels of ALT,AST and TBIL were significantly decreased on postoperative day l and 3,and the plasma concentrations of TNF-α and IL-6 were decreased on postoperative day 1,3 and 5 in group M.Conclusion Methylprednisolone can reduce hepatic I/R injury in the patients undergoing hepatolobectomy and inhibition of systemic inflammatory responses is involved in the mechanism.

Objective: This research explores the relationship between the immuno-suppression function of regulatory T cells (Treg) in the ascites of ovarian cancer (OC) patients, the clinico-pathologic features of these patients, and the correlation of the function of Treg with initial treatment and relapse status of the patients to further investigate the specific mechanism of immuno-regulatory func-tion of CD4+ CD25+ Treg in the ascites of OC. Methods: Immuno-magnetic activated cell sorting (MACS) was conducted to sort CD4+CD25+Treg and autologous CD4+CD25-Treg from the ascites of 28 OC patients. Carboxyfluorescein-diacetate succinimidyl ester (CFSE) was used to label the autologous CD4+CD25-Treg. These labeled cells were then used as controls and co-cultured with autologous CD4+CD25+Treg at the ratio of 1∶1 or 1∶2. The mean inhibition ratio of Treg in specimens to the proliferation of autolo-gous CD4+ CD25-Treg was calculated after the flow cytometry of the CFSE expression and Modfit software analysis of the CD4+CD25-Treg proliferation index (PI) were performed. Anti-IL-10 and/or anti-TGF-β1 antibodies were neutralized to investigate whether the CD4+CD25+Treg-mediated immuno-suppression escaped through the ascites can produce a marked effect by the inhibitory cyto-kine IL-10 or TGF-β1. Results: The mean inhibition ratio of CD4+ CD25- Treg in the ascites of stage Ⅲ to Ⅳ OC patients was (75.72±17.04)%, which is significantly higher than that of stageⅠtoⅡOC patients (59.61±16.97)%;P<0.05. In addition, Treg in the as-cites of OC patients with recurrent disease showed a significantly higher inhibition ratio than that of patients with primary disease;P<0.001. Moreover, Treg in groups added into neutralizing anti-IL-10 and/or anti-TGF-β1 antibodies displayed significantly lower depres-sant effect than the control group;P<0.05. Conclusion:The immuno-suppression of CD4+CD25+Treg in the ascites of OC patients is correlated with the tumor staging and status of the primary or recurrent diseases. Moreover, Treg may indicate a suppressor function by secreting cytokine IL-10 and TGF-β1.