Objective: In order to investigate the effects of activating and blocking Stat3 signaling pathway on invasion ability of human pancreatic cancer cells and explore its action mechanism.Methods:Human pancreatic cancer Capan-2 cells were treated with IL-6. SW1990 human pancreatic cancer cells were treated with AG490. Cell proliferation was measured by MTT assay. Western blotting and immunocytochemistry were performed to detect expression of phosphorylated Stat3 (p-Stat3) protein. Real-time fluorogentic quantitative PCR (RFQ-PCR) and Western blotting were used to detect the mRNA and protein expression of VEGF and MMP-2 mRNA, respectively. The invasion abilities of SW1990 and Capan-2 cells were determined by cell invasion assay in vitro. Results:IL-6 stimulated the proliferation of Capan-2 cells (P<0.05), elevated the expression of p-Stat3, increased the mRNA and protein expressions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 2 (MMP-2) (P<0.05), and enhanced the invasion ability of Capan-2 cells. AG490 inhibited the proliferation of SW1990 cells (P<0.05), down-regulated the expression of p-Stat3, markedly decreased the mRNA and protein expression of VEGF and MMP-2 (P<0.05), and weakened the invasion ability of SW1990 cells. Conclusion:Stat 3 signaling pathway plays an important role in the invasion and metastasis of pancreatic cancer. Stat 3 signaling transduction pathway may provide a novel therapeutic target for the treatment of pancreatic cancer.

objective To identify the three strains of pink-pigmented bacteria isolated from ticks.Methads 16S rDNA sequence analysis,16S rDNA similarity searching,bacterial morphological,physiological and biochemical tests and fatty acids analysis were used.Results The bacterial strains were pinkpigment,gram negative,strictly aerobic coccobacillus.They grow optimally at 37℃,while not in the presence of≥5% NaCl.The tests showed positive result for urease,D-glucose,citrate,UV absorption,and catalase.The predominant fatty acids were C16:0,C17:0,C18:1 ωgc and C19:0 cyc.Comparative 16S rDNA sequences analysis revealed that the strains were closely related to Roseomonas cervicalis and could reach up to 99.1% identity.Conclusion Based on phenotypic,biochemical and phylogenetic data,we propose these three isolates represent distinct species in genus Roseomonas,of which ticks may serve as a potential vector.

Objective To investigate the effect of RNAi-mediated sTAT3 gene silencing on the invasiveness of human pancreatic cancer cells. Methods STAT3 shRNA expression vector was stably transfected to SW1990 cells.STAT3 mRNA and protein expression were examined using reverse transcription polymerase chain reaction(RT-PCR)and Western blot,respectively.The invasion ability of SW1990 cells was determined by cell invasion assay in vitro.RT-PCR and Westem blot were performed to detect the mRNA and protein expression of the MMP-2 and VEGF,respectively. Results mRNA and protein expression of STAT3 were inhibited significantly by stable transfection of STAT3 shRNA expressing vectors.STAT3 silence with RNAi significantly inhibited the invasion ability of SW1990 cells decreasing protein and mRNA expression of MMP-2 and VEGF in SWl990 cells. Conciusion STAT3 silence with RNAi significantly inhibits the invasion ability of pancreatic cancer cells through down-regulating MMP-2 and VEGF.

Objective To investigate the effect of interleukin 6 (IL-6) on the growth and proliferation of human pancreatic cancer cell line Capan-2 and the signal transduction pathway. Methods MTr method was used to detect the effect of IL-6 of different concentrations on the growth and proliferation of Capan - 2 cells; cell apoptosis was detected by flow cytometry; the intracellular localization of phosphorylated STAT3 (P-STAT3) was determined by immunocytochemistry and western blot were used to detect P-STAT3, bcl-xl and Cyclin D1 in Capan-2 cells stimulated by IL-6. Results IL-6 (100 ng/ml) could remarkably promote the growth of Capan-2 cells from 1 to 4. 965 ± 0. 18 (P < 0. 05) ; the percentage of apoptosis decreased significantly from (3.21 ±0.23)% to (1.98 ±0.67)% (P <0.05) ; the expressions of P-STAT3, bcl-xl, Cyelin D1 increased significantly (P < 0.05), and the expressions of bcl-xl was positively correlated with that of P-STAT3 (r =0.985, P =0.015) ; the expressions of Cyclin DI was also positively correlated with that of P-STAT3(r=0.914,P=0.036),Conclusions IL-6 activate JAK/STAT signal transduction pathway,which played an important role in the growth and proliferation of Capan-2 cells in the presence of IL-6.

Objective To detect the impact of reactive oxygen species ( ROS) on mitochondrial morphology and distri-bution during mitosis.Methods A viral vector in which the fluorescence gene was specifically under the control of mito-chondrial promoter was constructed and confirmed through DNA sequencing and Western blotting.After transfecting HeLa s3 cell with packaged virus, the HeLa s3-COX4tp-EGFP cell line stably expressing the mitochondrial fluorescence signal was obtained.With immunofluorescent staining, the impact of ROS on the morphology and distribution of mitochondria dur-ing mitosis was inspected.Result The cell line constantly expressing mitochondrial fluorescence signals was successfully constructed.Meanwhile,it was found that H2 O2 treatment could significantly change the morphology and distribution of mi-tochondria during mitosis by confocal microscopy.Conclusion Our study demonstrates that ROS can affect the morphology and distribution of mitochondria during mitosis.This research help study the relationship between the mitochondrial function and the regulation of mitosis in the future.

Objective To investigate the perioperative characteristics and changing trends of gastric electrical activity of thoracic (tube) stomach in patients with esophageal cancer.Methods The clinical data of 30 patients with esophageal cancer who were admitted to the Sichuan Cancer Hospital between March 2013 and November 2013 were prospectively analyzed.All the eligible patients underwent esophageal cancer resection by Ivor-Lewis according to the inclusion criteria.The electrogastrograms of patients were recorded at preoperative day 1 and at postoperative day 3,7, 11 and 30.The electrogastrograms of patients at preoperative day 1 were used as the control.The parameters of electrogastrogram were analyzed including main frequency, coefficient of dominant frequency instablility, main power, postprandial/preprandial power ratio, percentage of normal gastric slow wave,percentage of slow gastric slow wave and percentage of tachycardia gastric slow wave.Measurement data with normal distritution were presented as x ± s, and measurement data with skew distritution were presented as M (Qn).The postoperative time and pre-and post-prandial electrogastrograms were compared by the repeated measures two-way ANOVA.The comparison between groups were evaluated with the LSD test and analysis of variance.Results Thirty patients were screened for eligibility with a mean age of 62 years (range, 49-75 years), including 26 males and 4 females.The pre-and post-prandial main frequencies were changed from 2.83 ± 0.13 and 3.01 ± 0.17 before operation to 2.66 ± 0.10 and 2.82 ± 0.10 at postoperative day 30 with coherent changing trend.The main frequencies at postoperative each time points were significantly lower than those before operation while postprandial above indicators were higher than preprandial those, showing a significant increasing trend with the passage of postoperative time (F =285.62, P ＜ 0.05).There was no interaction between the time and meal (F =0.22, P ＞ 0.05).The pre-and post-prandial coefficients of dominant frequency instablility were changed from 0.133 ±0.031 and 0.045 ±0.019 before operation to 0.150 ±0.043 and 0.115 ±0.010 at postoperative day 30 with coherent changing trend, and coefficients of dominant frequency instablility at postoperative each time points were significantly higher than those before operation while postprandial above indicators were lower than preprandial those, showing a significant reducing trend with the passage of postoperative time (F =16.51, P ＜ 0.05).The pre-and post-prandial main powers were changed from (85 ± 15) μV and (149 ± 23) μV before operation to (74 ± 9) μμV and (98 ± 10) μV at postoperative day 30, and main powers at postoperative each time points were significantly lower than those before operation, showing a significant increasing trend with the passage of postoperative time (F =48.45, P ＜ 0.05).There was interaction between the time and meal (F =7.39, P ＜ 0.05).The postprandial/preprandial power ratio was changed from 3.00 ± 0.35 before operation to 2.52 ± 0.25 at postoperative day 30, and postprandial/preprandial power ratios at postoperative each time points were significantly lower than those before operation, showing a significant increasing trend with the passage of postoperative time (F =26.66, P ＜ 0.05).The pre-and post-prandial percentages of normal gastric slow wave were changed from 81％ ± 6％ and 94％ ± 5％ before operation to 57％ ± 5％ and 70％ ± 5％ at postoperative day 30 with coherent changing trend, and percentages of normal gastric slow wave at postoperative each time points were significantly lower than those before operation while postprandial above indicators was lower than preprandial those, showing a significant increasing trend with the passage of postoperative time (F =49.36,P ＜0.05).There was no interaction between the time and meal (F =0.24, P ＞ 0.05).The pre-and postprandial percentages of slow gastric slow wave were changed from 17％ ± 7％ and 4％ ± 4％ before operation to 32％±4％ and 21％±4％ at postoperative day 30 with coherent changing trend, and percentages of slow gastric slow wave at postoperative each time points were significantly higher than those before operation while preprandial above indicators were higher than postprandial those, showing a significant reducing trend with the passage of postoperative time (F =46.54, P ＜ 0.05).There was interaction between the time and meal (F =18.12, P ＜ 0.05).The pre-and post-prandial tachycardia gastric slow wave percentages were changed from 1.55％ (1.04％,2.21％) and 1.95％ (1.74％, 4.22％) before operation to 8.97％ (5.76％, 12.02％) and 12.41％ (8.04％,16.85％) at postoperative day 30 without completely coherent changing trend, and percentages of tachycardia gastric slow wave at postoperative each time points were significantly higher than those before operation while postprandial above indicators were higher than preprandial those, showing a significant difference between before operation and postoperative day 3 (Z =11.47, 13.28, P ＜ 0.05) and no significant difference among the postoperative day 7, 11, 30 (Z =1.88, 0.31, 0.03, P ＞ 0.05).There was no interaction between the time and meal (F=0.85, P＜0.05).Conclusions After the esophagectomy, gastric electrical activity of thoracic (tube) stomach is also retained before and after the meal.There are significant differences among the main frequency, main power, coefficients of dominant frequency instablility, postprandial/preprandial power ratio,percentage of normal gastric slow wave, percentage of slow gastric slow wave, percentage of tachycardia gastric slow wave of thoracis (tube) stomach, they have changed dynamically in the perioperative period.

Objective To investigate the correlation between the expression of STAT3 and MMP-2 in human pancreatic cancer,and to probe the mechanism by which STAT3 signal pathway regulates the expression of MMP-2 in pancreatic cancer cells.Methods Immunohistochemistry was used to detect the expression of STAT3,phosphorylated STAT3(p-STAT3)and MMP-2 in pancreatic cancer tissues of 34 cases and normal pancreatic tissues of 10 cases.Correlation between the expression of STAT3、p-STAT3 and MMP- 2 were statistically analyzed.Human pancreatic cancer cell lines SW1990 was cultured.AG490,an inhibitor of the upstream Janus kinase(JAK)of STAT3 was added into the culture medium.Electrophoretic mobility shift assay(EMSA)was used to detect STAT3 DNA-binding activity in SW1990 cells.Western blot was used to detect the expression of STAT3,p-STAT3 in SW1990 cells.In addition,the protein and mRNA expression of MMP-2 in SW1990 cells were determined by Western blot and RT-PCR,respectively. Results Immunohistochemistry revealed that the expression rate of STAT3,p-STAT3 were both higher in pancreatic cancer tissues than in normal pancreas tissues(P

AIMTo explore the effect of cerebellar fastigial nuclei (FN)on lymphocyte function and the pathway mediating the effect.

METHODSKainic acid (KA) was microinjected into bilateral FN of rats to destroy neuronal bodies in the FN. On the eighth day after the surgery, lymphocyte percentage in the peripheral blood and level of sheep red blood cell(SRBC)-specific IgM antibody in the serum were measured by using blood corpuscle counter and enzyme-linked immunosorbent assay (ELISA), respectively.A technology of electrolytic lesion was used to destroy the projections of cerebellar FN neurons to hypothalamus in decussation of superior cerebellar peduncle(xscp).

RESULTSOn the eighth day after the microinjection of KA into the bilateral FN of rats, the Nissl-stained neuronal bodies in the FN disappeared and glia could proliferated within the damaged FN. In the nuclei close to FN, the interposed nuclei and the dentate nuclei, Nissl-stained neurons still could be seen. On the control cerebellar sections, in which FN was infused with saline, we could see the normal Nissl-stained neurons in the FN and the other two nuclei.On day 8 following the effective FN lesions, both the lymphocyte percentage in the peripheral blood and the level of anti-SRBC IgM antibody in the serum were significantly increased in comparison with those of control rats infused with saline in the FN. On the eighth day after electrolytic lesion of the fibres in xscp, the FN-hypothalamic projections were damaged and there were no visible BDA-positive endings in hypothalamus. Meanwhile, both the lymphocyte percentage in the peripheral blood and the level of anti-SRBC IgM antibody in the serum were remarkably enhanced relative to those of control rats with sham lesion of xscp.

CONCLUSIONThe electrolytic lesion of the FN-hypothalamic projections in xscp causes an enhancement of lymphocyte function similar to that of KA lesions of neuronal soma in the FN. These findings suggest that the cerebellohypothalamic projections participate in mediating the modulation of lymphocyte function by the cerebellum.

Animals ; Cerebellar Nuclei ; immunology ; injuries ; Female ; Hypothalamus ; immunology ; physiology ; Kainic Acid ; Lymphocyte Count ; Lymphocytes ; cytology ; immunology ; Male ; Neural Pathways ; immunology ; physiology ; Neuroimmunomodulation ; physiology ; Rats ; Rats, Sprague-Dawley

XYL1 gene,which encodes xylose reductase with dual coenzyme activity from Candida tropicalis,was transformed into Pichia pastoris X-33 by expression vector pGAPZB.The recombination strain was immobilized in Ca-alginate beads and fermentation characterization is studied using corn cob hydrolysates.Fermentation conditions were as follow:initial pH value 6.0,30℃,initial cell concentration of 20%,the Liquid volume of 28%,rotation speed 130r/min.The average xylitol yield was 37.5% on the optimum condition.This result is expected to provide a new alternative method for producing xylitol on a large scale by bioconversion.

Objective To discuss the long term clinical effect of ileal orthotropic neobladder.Methods From 1991 to 1998,79 patients,mean age 55(41～75)years,male 74,female 6,were followed up.The serum creatinine and urea,electrolytes,blood routine,B ultrasonic scan of the neobladder residual urine and IVU or MRU of the patients were followed up.The max transverse diameter of renal pelvis and the max verticaI/level diameter of neobladder were measured in 5,10 to 14,15 years of postoperative when IVU or MRU.All results of different time were compared by the multiple comparisons.The local or distant cancer recurrence and the complications of the operation Were evaluated. Results Sixty-four cases,58 male,6 famle,were long term followed up:mean time was 167 (range,121～216)months.Seven cases died of other diseases.Seven cases had pelvic recarrence.Two cases had urethral recurrence.Three cases died of tumor metastasis.One case had ureter recurrence.Forty-eight patients were alive more than 10 years.The value of the serum creatinine,urea,electrolytes and bloods routine of the patients were normal after 5,10 to 14 and 15 vears postoperative (P＞0.05).The max transverse diameter of the renal pelvis in 5,10 to 14 and 15 years Dostoperative were 14.0 mm,14.1 mm and 13.7 mm,respectively,P＞0.05.The max vertical/level diameter of the neobladder in 5,10 to 14,15 years of postoperative were 110.4 mm/90.4 mm,111.5 mm/95.3mm and 127.0 mm/97.0 mm,respectively,P＞0.05.The residual urine of 5 cases was more than 50 ml and had not increased during follow up.Eight cases with neobladder stone were cured by the intracavitary lithothrypsis.Two cases with uretheral stricture were cured by the intracavitary therapy.Twelve cases of 14 cases with inguinal hernia were cured by reoperation,2 cases accepted conservative treatment.Only 17 cases had no complication involve of the cancer and the operation. Conclusion The upper urinary tract and neobladder of the ileal orthotopic neobladder could be stable for long time,the cure rate of tumor is satisfactory and the lifetime follow up is necessary.