Objective To investigate the effect and mechanism of action in pancreatic cancer BxPC3 cell treated by cerulenin in combination with gemcitabine.Methods BxPC3 cells were cultivated with 10 μg/ml cerulenin,20 μmol/L gemcitabine or 10 μg/ml cerulenin + 20 μmol/L gemcitabine for 48 h,and cells without treatment were control.Cell proliferation was detected by CCK-8 assay,and early apoptosis was detected by AnnexinV/PI double staining method.The expression of Bcl-2 mRNA and Bax mRNA were detected by RT-PCR,and the protein level of Bcl-2,Bax were detected by Western Blot.Results The inhibition rate of BxPC3 cells were 0,(51.28 ± 1.84) ％,(53.59 ± 1.62) ％,(84.57 ± 1.01) ％ in control,cerulenin group,gemcitabine group,combination group; and the rate of early apoptosis was (0.83 ± 0.31) ％,(31.37 ± 1.04) ％,(38.33 ± 0.75) ％,(69.43 ± 0.83) ％,and the expression of Bcl-2 mRNA was 0.67 ± 0.01,0.44 ±0.01,0.36 ±0.08,0.27 ±0.07,and the expression of Bax mRNA was 0.14 ±0.01,0.31 ± 0.02,0.32 ± 0.03,0.91 ± 0.06 ; while the ratio of Bcl-2 mRNA/Bax mRNA was 4.78 ± 0.13,1.39 ± 0.04,1.15 ± 0.02,0.30 ± 0.02 ; the expression of Bcl-2 protein was 1.24 ± 0.04,0.51 ± 0.02,0.42 ± 0.02,0.13 ±0.01 ; and the expression of Bax protein was 0.20 ± 0.05,0.47 ± 0.01,0.54 ± 0.01,1.21 ± 0.03 ; while the ratio of Bcl-2/Bax was 6.00 ± 0.11,1.11 ± 0.01,0.77 ± 0.03,0.10 ± 0.06.And the difference among the groups was statistically significant (P ＜ 0.01).Conclusions Cerulenin combined with gemcitabin has synergistic effect on the inhibition of BxPC cells proliferation.Its mechanisms may be up-regulation of Bax,down-regulation of Bcl-2,and promoting apoptosis of cells through the decrease of Bcl-2/Bax ratio.

Objective To observe the clinical effect of Shuguanling(Chinese medicine for dredging the oviduct) on salpingemphraxis sterility and to study its mechanism.Methods The diagnosed patients were randomized into 3 groups with 52 in each.The treatment group was administered Shuguanling p.o.and clyster.The control group A was given Xuefu Zhuyu Oral Liquid(Oral liquid for removing blood stasis in the abdomen),and Kangfu Xiaoyan Pills(Pills for recovery of gynecological inflammation) suppository into recta.The control group B was prescribed Xuefu Zhuyu Oral Liquid and vaginal culdocentesis with the mixture of dexamethasone,chymotrypsin,gentamicin and normal saline.After three months,the effect was evaluated and the pelvic blood flow,blood viscosity,plasma collagenase,fibrinogen and fibrinolytic activity of the treatment group were tested for comparison.Results The tubal patency rate and pregnancy rate of treatment group were significantly higher than those of control groups(P

ObjectiveTo investigate the effect of thalidomide on the proliferation and apoptosis in human pancreatic cancer cell SW1990 in vitro.MethodsSW1990 cell line was treated with thalidomide at different concentrations (3.125,6.25,12.5,25,50,100,200 and 400 μg/ml) for 24,48,72 h,and then cell proliferation were evaluated by MTT.Cell cycle was determined by flow cytometry,apoptosis was determined by annexin V/PI fluos staining,Bcl-2,Bax protein expression and Bcl-2/Bax ratios were measured by Western Blot in vitro.ResultsThalidomide inhibited the proliferation of SW1990 cells in a time and dosedependant manner.The proportion of G0/G1 phase of SW1990 cells with 200 μg/ml thalidomide treatment increased from (41.15 ± 2.23 ) ％ to (58.83 ± 2.33 ) ％,apoptosis rate increased from 2.6％ to 28.0％,the expression of Bax protein up-regulated from 0.17 ± 0.03 to 0.33 ± 0.04,the expression of Bcl-2 protein downregulated from 0.35 ± 0.02 to 0.17± 0.01,the ration of Bcl-2/Bax decreased from 2.17 ± 0.44 to 0.52 ±0.07.ConclusionsThalidomide can inhibit the proliferation of pancreatic cancer SW1990 cells by upregulating Bax,down-regulating Bcl-2,inducing cell apoptosis and cell Go/G1 phase arrest.

Objective To evaluate the significace of endoscopic diagnosis of gastrointestinal graft-versus-host disease ( GI GVHD) resulted from allogeneic bone marrow transplantation. Methods Five patients with suspected GI GVHD received endoscopic examination and biopsies were taken from antrum, sig-moid colon or the focal lesions. Results The symptoms of GI GVHD included anorexia, nausea, vomiting, watery diarrhea, abdominal pain, GI bleeding etc. The endoscopic appearance in stomach varied from subtle mucosal edema, hyperemia, and erythema necrotic erosion. Besides the above stated, the endoscopic appearance in colon also presented with diffuse mucosal erosion, hemorrhege and ulcer. Histological findings included characteristically crypt epithelial cell apoptosis and sloughing, and lymphocyte infiltration in epithelium and lamina propria. The involvement may be varied from diffuse to focal in stomach or colon. Conclusion Endoscopic and histological evaluation of the stomach and colon and biopsy specimen can be used as the evidences in diagnosing GI GVHD.

Objective To explore the role of receptor‐interacting protein kinase 3 (RIP3)‐mediated necroptosis in ulcerative colitis (UC ) . Methods The colonic mucosa tissues of eighty‐five patients diagnosed with UC were collected .Disease staging and pathological classification of UC patients were evaluated according to Mayo standard and Truelove standard ,respectively .The expressions of RIP3 and tumor necrosis factor (TNF)‐α were detected by immunohistochemistry .The cell inflammation model in HT‐29 cells was induced by interferon (IFN)‐γ.The expression of RIP3 mRNA and interleukin (IL)‐1βmRNA was determined by real time polymerase chain reaction (RT‐PCR) .Twenty healthy individuals were studied as controls .A single factor analysis of variance was performed in the comparison among groups ,and Pearson correlation analysis was used for correlation analysis .Results Among 85 patients with UC , 27 cases were mild ,30 cases were moderate and 28 cases were severe;for pathological classification ,26 were gradeⅠ ,31 were gradeⅡ and 28 were grade Ⅲ .The Mayo score was positively correlated with pathological grade (r=0 .997 , P<0 .05) .The expression of RIP3 and TNF‐α in colonic mucosa tissues of UC group increased along with the pathological classification grade .The expression of RIP3 of gradeⅠto Ⅲ was 1 .81 ± 0 .98 ,2 .77 ± 1 .26 and 4 .86 ± 2 .77 ,respectively ,and the differences were statistically significant (F=26 .10 ,P<0 .05) .The expression of TNF‐αof gradeⅠto Ⅲ was 1 .35 ± 0 .69 ,2 .61 ± 1 .41 and 4 .43 ± 2 .17 ,respectively ,and the differences were statistically significant (F=31 .80 ,P<0 .05) .The expression of IL‐1β mRNA of IFN‐γ group ,IFN‐γ+ z‐VAD group and IFN‐γ+z‐VAD+ TNF‐αgroup was 0 .68 ± 0 .27 ,1 .47 ± 0 .12 and 1 .86 ± 0 .16 ,respectively ,and the differences were statistically significant (F=38 .45 , P<0 .05) .The expression of RIP3 mRNA of IFN‐γgroup ,IFN‐γ+z‐VAD group and IFN‐γ+z‐VAD+TNF‐αgroup was 0 .46 ± 0 .13 ,1 .21 ± 0 .29 and 2 .06 ± 0 .20 ,respectively , and the differences were statistically significant (F= 55 .15 , P< 0 .05) .Conclusions TNF‐α/RIP3 signal pathway is up‐regulated in UC .RIP3‐mediated necroptosis promotes the death of colonic epithelial cells and exacerbates inflammatory response .RIP3‐mediated necroptosis may be an important mechanism in UC .

Objective To study the protective effects of ulinastatin (UTI) on canine islets isolated by automated method and to determine whether the addition of UTI during the in situ pancreatic perfusion process may improve islet recovery. Methods There was no significant difference in donor-related factors (age, gender, warm ischemia time, cold storage time, weight and pancreas weight) between the two experimental groups. Twenty donors were randomly divided into two groups. Pancreases were in situ perfused via abdominal aorta using cold hypertonic citric potassium solution (HC-A) 2 500 ml, group 1 (HC-A group, n =10). The pancreases were perfused with cold HC-A and UTI ( 10 000 U/kg), group 2 (HC-A+UTI group, n =10). Intraductal collagenase V (4℃) delivery with controlled perfusion was done, the pancreas was dissociated in system of Ricordi Chamber and their purified islets were separated with Ficoll density gradient centrifugation. The digestion time, islets remaining trapped in exocrine tissue, final islet purity, insulin and C-pep secretory activity, and islet recovery were observed. The purified islets were observed under light microscopy and electronic microscopy. Results UTI supplementation did not affect the digestion time, islets remaining trapped in exocrine tissue, final islet purify, insulin and C-pep secretory activity and their ultrastructure, and islet recovery was increased. In the HC-A+UTI group the yield of islets was [( 6.17 ? 2.86 )?10 4] IEQ, while in the HC-A group, that was [( 3.42 ? 1.47 )?10 4] IEQ ( P

Objective To study the operative design, safety evaluation and feasibility of gasless laparoscopic operations. Methods The gasless laparoscopic devices (GLD) were applied to 124 cases of laparoscopic choledocholithotomy with T-tube drainage, 56 cases of laparoscopic choledocholithotomy with primary bile duct suture, 1 case of laparoscopic choledochocystectomy hepaticojejunostomy, 15 cases of modified gasless laparoscopic choledochojejunostomy, 1 case of hand-assisted laparoscopic megalosplenic resection and portozygos disconnection, and 1 case of gasless device assisted laparoscopic splenectomy. Results All the operations were successfully conducted, without any severe complications. Follow-up for 1~9 years in 129 cases of biliary duct operations found 1 case of recurrence. Follow-up for 3~12 months in 2 cases of splenic operations found no recurrence. Conclusions Gasless laparoscopic operations in this study have reached minimally invasive outcomes, proven to be safe and feasible.

Objective To study on the endoscopic, pathologic and immunohistochemical characteristics of gastric stromal tumor. Methods The gastric tumor samples were studied with light microscopy, and in those suspected of mesenchymal tumor the expression of CD11734, CD34 , vimentin, desmin, S-100 etc. were further determined with SP method. The clinical, pathological and endoscopic characteristics were reviewed. Results Twenty-eight cases of mesenchymal cell tumor were diagnosed as stromal tumor and enrolled in this study, with 9 benign, 11 borderline, and 8 malignant cases. Endoscopically, the gastric stromal tumors showed submucosal protuberant lesions, and were situated in fundus (15 cases), body (10 cases) and an-trum (3 cases). Deep concave ulcer presented on tumor surface in 10 patients. Histology revealed the typical stromal tumor features with cell in spindle and/or epithelia shape and arranged in fence-like or spiral forms. All cases were positive for CD117 CD34 , vimentin when assessed, but 28 cases were negative for desmin and 22 cases (18. 2% ) for S-100. Conclusion The gastric stromal tumors were the most common mesenchymal tumors. Microscopically, the tumors were composed of spindle or epithelioid cells. CD117 and CD34 can be used as immune markers to diagnose gastric stromal tumor.

Objective To investigate the effect of melatonin (MT) on the proliferation and apoptosis in human pancreatic cancer cell SW1990 in vitro.Methods SW1990 cell line were treated with MT at different concentrations (0.1,0.5,1.0,2.5 and 5.0 mmol/L) and at different time points (24 h,48 h and 72 h).Cell proliferation was evaluated by MTT and apoptosis was determined by AnnexinV/Pl,cell cycle was determined by flow cytometry,and Western Blot was used to detect the expression of Bcl-2 and Bax.Results MT could inhibit the proliferation of SW1990 cell in a time and dose dependent manner.48 h after 0.1～5.0 mmol/L MT treatment,the proliferation inhibitory rate was 7.4%～85.8%,the proportion of SW1990 cell in phase G0/G1 was 72.6%～85.33%.48 h after 1～5.0 mmol/L MT treatment,the apoptosis rate was 21.5% ～41.7%,and the expression of Bcl-2 was down-regulated and the ratio of Bcl-2/Bax decreased.Conclusions MT could inhibit the proliferation of SW1990 pancreatic cancer cells by up-regulating the expression of Bax and down-regulating the expression of Bcl-2,as well as enhancing apoptosis and blocking SW1990 cell in phase G0/G1.

Objective To investigate the therapeutic effects of melatonin (MT) on human pancreatic cancer in Balb/c nude mice subcutaneous xenograft model. Methods Pancreatic cancer model was established high dose MT(20 mg· kg -1 · d-1 ) , gemcitabine( GEM,50 mg· kg -1 · d-1 ) and high dose MT combination with GEM. Tumor size was measured regularly. The tumor growth curve was drawn. The activity of mice spleen natural killer cell was determined by MTT. Results Compared with the controls [ ( 1. 476 ± 0.075) cm3 ], tumor volumes in low dose MT group[ (0.998 ±0.112)cm3], high dose MT group[ (0.756 ±0.128) cm3], GEM group [ (0. 746 ± 0. 115 ) cm3 ], combination group [ (0. 305 ± 0. 111 ) cm3 ] were significantly decreased ( P <0.01), and the tumor size in high dose MT group was smaller than that in low dose MT group, while the tumor size in combination group was the smallest. The activity of mice spleen natural killer cell was ( 18.07 ± 1.23) %in control group, and they were (44.27 ±3.19)% ,(45.16 ±3.20)% and (30.29 ±2.91)% in low dose MT group, high dose MT group, combination group, which were significantly higher than those in the control group;the activity of natural killer cell in GEM group was ( 14.24 ± 2.70) %, which was significantly lower than that in the control group (P <0.05), but the activity of natural killer cell in combination group was significantly higher than that in the GEM group (P < 0.01 ). Conclusions MT could improve the immune function of mice,inhibit the growth of tumor, and MT combination with GEM may have more potent antitumor effect.