OBJECTIVE: To identify and analyze drug sensitivity of Corynebacterium glucuronolyticum iscolated from clinic, and to provide reference for clinical drug use. METHODS: Two strains isolated from the urine specimens of urolithiasis-induced urinary tract infection patients in our hospital were inoculated into Columbia blood plate and the MacConkey plate. The growth of strains was observed and counted. Protein mass spectrometry of strains was detected by MALDI-TOF-MS. DNA of strains was extracted, and PCR was used to amplify the 16S ribosome RNA (rRNA) sequence. Bi-directional sequencing of 1 500 bp target bands was conducted. Blast comparison between it and GenBank database was conducted to identify bacterial strain. Drug resistance of 2 strains was monitored by Etest assay. RESULTS: Two strains grew on the Columbia blood plate (with colony forming unit ＞105 CFU/mL) and did not grow on the MacConkey plate. Two strains were Gram-positive Corynebacterium and showed palisading or eight type arrangement. Two strains were C. glucuronolyticum by MALDI-TOF-MS identification, with reliability of 99. 9％. The characteristic peaks of m/z 2 431, 3 089, 3 364, 3 378, 4 200, 5 508, 6 302, 6 637, 6 730, 6 946, 12 603 appeared. Blast comparison showed that the sequence homology of 2 strains compared with C. glucuronolyticum strain known in GenBank were higher than 98 ％. Results of drug sensitivity test showed that strain 1 was resistant to ceftriaxone and ciprofloxacin, and sensitive to 14 other antibiotics as penicillin G; strain 2 was resistant to ceftriaxone, erythromycin, ciprofloxacin, tetracycline and clindamycin, moderately sensitive to cefotaxime, and sensitive to 10 other antibiotics. CONCLUSIONS: Two strains are C. glucuronolyticum, and drug resistance of them to commonly used antibiotics is different. The strains are rare pathogen of urinary tract and show multidrug resistance. Antibiotics should be selected according to the results of strain identification and drug sensitivity test.

In view of gemcitabine resistance has limited clinical activity of gemcitabine as a cellulotoxic drug in pancreatic cancer patients, this study is designed to investigate the effect of emodin on the sensitivity of pancreatic cancer to gemcitabine as well as its mechanism. After gemcitabine-resistant pancreatic cancer cell line (SW1990/GZ) was established by escalating doses of gemcitabine serially in pancreatic cancer cell line (SW1990). The cellular proliferation was detected by cell counting kit-8 (CCK-8) assay. Flow cytometry (FCM) was used to determine apoptosis of pancreatic cancer cells. The activity of NF-kappaB in pancreatic cancer cells was measured by electrophoretic mobility shift assay (EMSA). Western blotting was used to detect the protein expression of Bcl-2 and Survivin in SW1990/GZ cells. Metastatic model simulating human pancreatic cancer was established by orthotopic implantation of histologically intact human tumor tissue into pancreatic wall of nude mice. Also, immunohistochemistry was used to detect the positive expression of Ki-67, NF-kappaB, Bcl-2 and Survivin in the tumors. The results show that pretreatment of cells with emodin followed by gemcitabine induced a higher percentage of growth inhibition and apoptosis of pancreatic cancer cells than that of gemcitabine alone. In addition to in vitro results, emodin in combination with gemcitabine is much more effective as an antitumor agent compared to either agent alone in the orthotopic tumor model. Further study showed that the emodin with or without gemcitabine significantly down-regulates NF-kappaB and its regulated molecules such as Bcl-2 and Survivin proteins both in vitro and in vivo. It is concluded that inactivation of NF-kappaB signaling pathway by emodin resulting in the chemosensitization of pancreatic cancer to gemcitabine, which is likely to be an important and novel strategy for the treatment of pancreatic cancer.

OBJECTIVETo investigate the anti-metastasis effect of emodin on the pancreatic cancer in vitro and in vivo.

METHODHuman pancreatic cancer cell line SW1990 was treated with different concentrations of emodin (10, 20, 40 micromol x L(-1)) for 2 h, the effects of emodin on the migration and invasion of SW1990 cells were examined by using wound assay and matrigel counting. Western blot was used to detect the protein expression of NF-kappaB and MMP-9 in SW1990 cells after various concentrations of emodin (10, 20, 40 micromol x L(-1)) treatment for 48 h. Metastatic model simulating human pancreatic cancer was established by orthotropic implantation of histologically intact human tumor tissue into pancreatic wall of nude mice, and then divided into three groups: control group, low-dose emodin group (L-EMO) and high-dose emodin group (H-EMO). Eight weeks after implantation, the presences of metastasis were evaluated respectively after the mice were sacrificed. Immunohistochemistry was used to detect the positive expression of CD34, NF-kappaB and MMP-9 in the tumors.

RESULTEmodin suppressed the migration and invasion of SW1990 cells in a dose-dependent manner. Western bolt assay indicated that emodin down-regulated the expression of NF-kappaB and MMP-9 proteins in SW1990 cells. The incidences of metastasis were decreased significantly in L-EMO group and H-EMO group as compared with that in control group. The percentage of CD34, NF-kappaB and MMP-9-positive cells in the tumors were significantly reduced by the administration of emodin.

CONCLUSIONEmodin exerts anti-metastatic activity in pancreatic cancer both in vitro and in vivo, which may be related to down-regulation of NF-kappaB and MMP-9.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Line, Tumor ; Emodin ; pharmacology ; Female ; Humans ; Matrix Metalloproteinase 9 ; analysis ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; analysis ; antagonists & inhibitors ; Neoplasm Metastasis ; prevention & control ; Pancreatic Neoplasms ; blood supply ; drug therapy ; pathology

OBJECTIVETo evaluate the enhanced effect of gemcitabine by emodin and the possible mechanisms of the enhancement.

METHODBased on the model of SW1990 cell xenograft on athymic mouse, the mice were randomized to four groups with intraperitoneal (IP) injections of different drugs: group N (injecting 0.9% sodium chloride), group E (emodin, 40 mg x kg(-1)), group G (gemcitabine, 125 mg x kg(-1)), and group E + G (emodin 40 mg x kg(-1) and gemcitabine 80 mg x kg(-1) in combination). The tumor volume, tumor weight and body weight of mice were measured during the drug therapy. The mice were sacrificed one week after last injection of drug. Tunel assay were used used to detect the apoptosis of tumor cells. And immunohistochemistry (IHC) and Western blot (WB) were used to detect the variance of the apoptosis relative protein expression of Bax, Bcl-2, and Cytochrome C .

RESULTOne week after the last administration, the mean tumor volume and tumor weight in group E + G were significantly decreased compared to the other groups. Tunel assay showed group E + G presented apparently more apoptosis than the other groups. Immunohistochemistry (IHC) and Western blot (WB) analysis showed the expression of Cytochrome C in cytoplasmin and Bax in group E + G was apparently upregulated while the expression of Bcl-2 was apparently downregulated compared to the other groups. As a result, Bcl-2/Bax ratio was significantly decreased in group E + G.

CONCLUSIONEmodin can significantly improve the antitumor effect of gemcitabine on transplanted tumor of SW1990 cell line through apparently enhancing the tumor cell apoptosis by gemcitabine. Downregulation of Bcl-2/Bax ratio and promoting release of Cytochrome C from mitochondria is possibly one of the mechanisms of the augmented apoptosis.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Cytochromes c ; metabolism ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Drug Synergism ; Emodin ; pharmacology ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Mice ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

Objective To study the effects of arsenic trioxide ( As2 O3 ) on inhibiting the proliferation of pancreatic carcinoma cell lines. Methods The inhibiting rate of As2O3 and Fluorouracin(5-Fu) ,Gemcitabine(GEM) on pancreatic carcinoma cell lines PC-3 were detected by using CCK-8 assay. Results As compared with 5-Fu、GEM,the inhibiting rate of As2O3 was the highest one( P ＜ 0.01 or P ＜ 0.05). Conclusion As2O3 can inhibit pancreatic carcinoma cell lines PC-3 effectively in vitro. The effects of As2O3 on inhibiting the proliferation of pancreatic carcinoma cell lines was stronger than 5-Fu and GEM. This is possibly due to the extensive and unique anticancer mechanism of As2O3.

OBJECTIVETo investigate the mechanism of Emodin on the role of acute rejection in rat liver transplantation.

METHODForty-eight pairs of orthotopic liver transplantation model were established with inbred rats which were randomly divided into 3 groups: Control group (BN --> BN), acute rejection group (Lewis --> BN) and emodin group (Lewis --> BN). Six recipients in each group were randomly collected and contents of TNF-alpha and IL-10 in the peripheral blood were detected with ELISA on Day 1, 3, 5 and 7 separately after transplantation and histopathological evaluation was made to detect the differences among groups after the livers were taken out on day 7. The other 10 in each group were protected to evaluate the animation and life time.

RESULTThe average meso-life time in emodin group (25.6 days) is significantly longer (P < 0.05) than acute rejection group (10.9 days). Compared with the acute rejection group, Emodin group shows up less rejection in the histopathological evaluation (P < 0.01), less TNF-alpha (P < 0.05) and a significant up-regulation of IL-10 in the peripheral blood (P < 0.05 after day 3).

CONCLUSIONEmodin can inhibit the acute rejection of liver transplantation in rats model effectively and it may play the role with reduction of TNF-alpha and upregulation of IL-10.

Animals ; Emodin ; pharmacology ; Gene Expression ; drug effects ; Graft Rejection ; drug therapy ; Interleukin-10 ; genetics ; immunology ; Liver Transplantation ; immunology ; Male ; Random Allocation ; Rats ; Rats, Inbred Lew ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; genetics ; immunology

Objective:To evaluate the effect of emodin on acute rejection after orthotopic liver transplantation(OLT) in rats.Methods:The LEW→BN OLT models were established.A total of 45 rats were divided randomly and equally into 3 groups:group A was treated with normal saline at dose of 0.5 ml/d intraperitoneally from 1st day to 8th day after operation;Group B,CsA at dose of 10.0 mg?kg-1?d-1;Group C,emodin at dose of 50.0 mg?kg-1?d-1.8 days after operation,6 recipients of each groups were killed for confirming rejection-active index(RAI) and hepatocellular apoptosis index(AI) by observing the pathologic change of transplanted liver in recipients.The other recipients were raised for observing the survival time.Results:Respectively,the survival time(days) of group A,B,C was 9.50?1.64,21.57?2.15,21.29?2.21.The survival time of group B,C was significantly longer than that of group A(P0.05).Respectively,the RAI of group A,B,C was 7.67?0.9,5.17?0.40,5.83?0.75 and the AI of group A,B,C was 35.83?2.32,15.83?1.33,16.50?2.35.The RAI and AI of group B,C was significantly lower than that of group A(P0.05).Conclusion:Emodin can reduce the hepatocellular apoptosis and suppress the acute rejection after OLT in rats.

Objective: To investigate the effect and mechanism of emodin (EMO) on human pancreatic cancer cell line BxPC-3 in vivo. Methods: After the pancreatic cancer model in nude mice was established, the mice were divided into four groups: control group (NS 0.2mL/d by i.p. injection), EMO group (EMO 40mg?kg-1?d-1 by i.p. injection), and Gemcitabine (GEM) group (GEM 80mg/kg, twice/week by i.p. injection) with 8 mice each group. After 2 weeks of administration, the mice were sacrificed, detected the body-weight change of nude nice before and after the experiment, and recorded the growth inhibition rate of tumor (TGI). Immunohistochemical (IHC) staining for ki-67 and terminal deoxynucleotidyl transferase-mediated nicked labeling assay (TUNEL) were undertaken to detect the cell proliferation and cell apoptosis in tumor tissue in xenograft nude mice. Results: The inter-group comparisons in body-weight of nude mice showed no significant difference in comparing group NS(27.0?1.64)g with group EMO(25.1?1.58)g and GEM(25.6? 1.47)g.The EMO group was 38.46%, the GEM group was 44.23%. The inter-group comparisons in immunohistochemical analysis of ki-67 showed significant difference in comparing group NS IOD(219.5?17.98) with group EMO IOD(146.6? 11.57)and GEM IOD(139.5?12.55), (P

Objective To improve the diagnosis and treatment of liver trauma. Methods We retrospectively analyzed different therapeutic means on liver trauma including 79 cases with operative treatment and 27 cases with nonoperative treatment. Results In nonoperative treatment group, 25 cases were cured, 2 cases died. In operative treatment group, 73 cases were cured, 6 cases died. Cure rate was 92.5%(98/106), mortality was 7.5%(8/106). 5 cases died of major blood vessels rupture, 3 cases died on multiple organ function failure. Postoperative complications included 3 cases of subphrenic infection, 10 cases of hepatic abscess, 8 cases of pleural hydrops, 7 cases of incisional infection. Conclutions Type I of liver trauma can be treated by nonoperation, type Ⅱ～Ⅳ of blunt liver trauma can be treated by nonoperation on the condition of intensive monitoring. Type Ⅱ～ Ⅵ of liver trauma should be operated on emergently in case of massive intraabdominal bleeding and combined organ injury.

Objective To explore the prevention and treatment of lymphorrhagia due to lymph node dissection in gastric carcinoma. MethodsClinical data of 743 patients undergoing radical gastrectomy plus lymphadenectomy from January 1997 to March 2004 were analyzed retrospectively. ResultsThe patients in D3 or D4 lymphadenectomy suffered from a higher lymphorrhagia rate than those in D2 lymphadenectomy(P