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Objective To investigate the therapeutic effects of cFLIP-siRNA tail venous injection on acute necrotizing pancre.atitis (ANP) rats,and explore the related mechanism.Methods Three pairs of cFLIP siRNAs (cFLIP-siRNA) and negative control(siRNA-NC) were designed and synthesized.The most effective siRNA to inhibit the expression of cFLIP gene was selected for the experiment.Thirty SD rats were treated with retrograde pancreaticobiliary duct injection of 5％ taurocholic acid sodium salt solution to establish ANP rat model.They were randomly divided into ANP group,siRNA-NC group and cFLIP-siRNA group with l0 rats in each group using random number method.The same volume of normal saline,siRNA-NC and cFLIP-siRNA were injected into tail vein,respectively.The rats were sacrificed after 24 h and the blood samples were colelcted.The levels of serum amylase and endotoxin were measured.The pancreatic tissues were collected and routine pathological examination was performed.The expressions of cFLIP-S and caspase-8 protein in pancreatic tissues were detected by immunohistochemistry and Western blotting.Results The levels of serum amylase in ANP group,siRNA-NC group and cFLIP-siRNA group were (1 286 ± 209),(1 297 ± 305) and (552± 256)U/L,the level of serum endotoxin were (136± 32),(128± 56) and (46± 23)ng/L,respectively.The pancreatic pathological scores were (4.97 ± 1.16),(4.92 ± 2.03) and (1.67 ± 1.98).The expression level of cFLIP-S protein were (8.56 ± 2.72),(9.12 ± 2.45) and (3.82 ± 1.46),respectively,which were significantly lower in cFLIP-siRNA group than in ANP group and siRNA-NC group (P ＜ 0.05),and the difference was statistically significant (P ＜ 0.05).The expression level of caspase-8 protein in the three groups were (2.25 ± 1.24),(2.41 ± 1.14) and (5.56 ± 1.79),respectively,which were significantly higher in cFLIP-siRNA group than in ANP group and siRNA-NC group,and the difference was statistically significant (P ＜0.05).However,there was no significant difference between ANP group and siRNA-NC group.Conclusions Inhibition of cFLIP gene expression can reduce pancreatic injury in ANP rats.The possible mechanism was that the up-regulation of caspase-8 can inhibit pancreatic cell necrosis and promote apoptosis.
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Objective To investigate the significance of fibrin monomer(FM ) ,D‐Dimer(D‐D) and other blood coagulation indica‐tors detection in the early diagnosis of disseminated intravascular coagulation(DIC) in obstetrics .Methods 90 healthy women with‐out pregnancy(control group) ,270 normal pregnant women including early ,mid‐,late pregnancy(90 cases for each) ,36 early DIC puerperas(early DIC group) were enrolled in the study .FM ,D‐D ,prothrombin time (PT ) ,activated partial coagulation activity (APTT) ,fibrinogen(FIB) levels were detected for those people .Results The levels of D‐D and FM in early DIC group were signif‐icantly higher than those in control group (P<0 .05) ,and PT ,APTT in late pregnant group were significantly shorter than those in early ,mid‐pregnant group and control group ,while FIB was significantly higher ,the difference was statistically significant(P<0 .05) .Conclusion PT ,APTT and FIB can reflect the high blood coagulation state in the late pregnancy ,but can not be used in the diagnosis of early DIC .D‐D and FM have important value for the early diagnosis of DIC in obstetrics .
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Objective To evaluate the value of urinary normal and modified nucleosides in the diagnosis of primary gallbladder carcinoma.Methods Between March 2011 to September 2012,28 patients with primary carcinoma of the gallbladder (PCG) were included in this study.Spontaneous urine samples were collected and 10 kinds of urinary nucleosides in the sample were determined by reversed-phase high performance liquid chromatography method.Another 62 patients with benign gallbladder disease and 70 healthy volunteers were enrolled as controls.Routine clinical tumor markers,including serum CA19-9 and CA125 levels of the PCG patients were also evaluated.Results 10 kinds of nucleoside had a good linear correlation (r＞0.99),and the recovery rate was between 87.4％ ～121.5％ range,and the accuracy rate was between 87.7％ ～121.3％,and the RSD of precision was between 1.5％～10.5％ range.In addition to adenosine and cytidine,the mean levels of the rest of the urinary nucleosides in the PCG group were much higher than those of the controls (P＜0.01).Based on principal component analysis,89.3％ of the PCG patients were correctly identified,which was much higher than those detected by CA19-9 (60.7％) and CA125 (67.9％) (P＜0.01).Conclusion Urinary nucleosides may become additional tumor markers which when combined with other clinical methods may help in the screening and early diagnosis of primary gallbladder cancer.
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Objective Epithelial mesenchymal transition (EMT) has a role in the proliferation and metastasis of various types of cells.This study investigates the hepatic oval cell's mechanism of EMT induced by histone deacetylase inhibition and the resulting cell motility increase from EMT.Methods Hepatic oval cell stem cell markers and marker changes were detected by flow cytometry,and after histone deacetylase inhibition induced EMT,the morphological changes were recorded.Western blot and quantitative real-time polymerase chain reaction detected the expression of E-cadherin,vimentin and Snail.Furthermore,confocal microscopy analysis recognized the nuclear translocation of Snail.Results Flow cytometry revealed no changes in the stem cell properties of hepatic oval cells in the cell culture process.Oval cell EMT,induced by HDACi,was observed through morphological changes,a reduction of the epithelial cell marker E cadherin,and an increase of the mesenchymal cell marker Vimentin.HDACi can promote the expression and nuclear translocation of Snail,which is the key hepatic oval cell transcription factor for both EMT and enhanced motility.Therefore,Snail RNA interference can suppress HDACi induced EMT in hepatic oval cells.Conclusions In conclusion,histone deacetylase inhibition induces hepatic oval cell epithelial-mesenchymal transition by Snail.
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ObjectiveTo investigate the effects of 5-Aza-CdR on the transcriptional regulation through methylation of the DNA promoter protocadherin 8(PCDHg) gene in pancreatic cancer cell line Capan-2.The Capan-2 retardation in growth rate and apoptosis were assessed in when administered 5-Aza-CdR and the chemotherapy agent,gemcitabine.MethodsMTT and flow cytometry were used to analyze the cell growth inhibition and apoptosis when treated with 5-Aza-CdR or in combination with gemcitabine.Methylation-specific PCR,RT-PCR and western blot were performed to detect methylation state,mRNA and protein respectively of PCDH8 gene in 5-Aza-CdR-treated Capan-2cells.Results Capan-2 cells treated with 5-Aza-CdR showed a slower growth rate,and a significant growth inhibition when given both 5-Aza-CdR in combination with gemcitabine.Compared with single drug administration and control,5-Aza-CdR together with gemcitabine can induce a stronger apoptosis signal.Different concentrations 5-Aza-CdR of were able to reverse methylation,restore mRNA and protein levels of PCDH8 in Capan-2.Conclusion5 Aza-CdR may demethylate the PCDH8 gene,which would effectively remove the gene silencing caused by high methylation,and thus induce gene mRNA transcription and protein expression to inhibit cell growth and have collaborative antitumor functions with gemcitabine.
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Objective To investigate the efficiency and safety of laparoscopic hepatectomy (LH) and open hepatectomy (OH) in patients with hepatocellular carcinoma (HCC). Methods A total of 45 HCC cases undergoing hepatectomy from January 2008 to December 2010 in our hospital were divided into LH group ( n =21 ) and OH group ( n =24 ).Their clinical efficiency and safety were compared and analyzed. Results The amount of intraoperative blood loss and blood transfusion in LH group were significantly higher than that of OH ( respectively x2 =3.973,x2 =4.862,all P ＜ 0.05 ).LH is associated with less postoperative complications and duration of hospital stay,compared with OH ( respectively x2 =4.746,t =-2.717,all P ＜0.05).Cumulative survival rates in both groups were not statistically significant ( x2 =0.172,P ＞ 0.05 ).However tumor-free survival rate in LH was significantly higher,superior to that of OH (x2 =4.543,P ＜ 0.05). Conclusions LH and OH are all safe and effective hepatectomy for HCC.LH has the advantage of less complication and shorter hospital stay.
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Objective This retrospective study was to explore the efficacy and determine the risk factors of survival for recurrent hepatocellular carcinoma ( HCC) treated by repeat hepatectomy. Methods From January 1995 till December 2010, 60 patients with recurrent HCCs, were treated by repeat hepatectomy.The significance of seventeen clinical or pathological variables in the risk factors of overall survival were assessed. Results The overall survival 1,3, and 5-year survival rates were 76. 3% , 40.7% and 25. 0% (from repeat hepatectomy), and 95. 0% , 62. 6% and 43. 3% ( from initial hepatectomy) , respectively.Univariate analysis indicated that tumor size at initial hepatectomy, recurrence interval from initial hepatectomy, serum albumin(ALB) level, resection margin, diameter of largest recurrence tumor and rumor vascular invasion were significant prognostic factors(P <0. 05, Kaplan-Meier Log-rank test). Multivariate analysis showed recurrence interval from initial hepatectomy, resection margin, diameter of largest recurrence tumor and rumor vascular invasion were significant prognostic factors(P<0.05, Cox proportional hazards model).Conclusion Repeat hepatectomy is effective for recurrent HCC. Recurrence interval from initial hepatectomy, resection margin, diameter of largest recurrence tumor and rumor vascular invasion were significant prognostic factors.
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Objective To compare the efficacy of percutaneous radiofrequency ablation (PRFA) and repeat hepatectomy for solitary recurrent hepatocellular carcinoma (HCC) with the diameter≤3 cm. Methods The clinical data of 151 patients with recurrent HCC (diameter≤3 cm) who were admitted to the Cancer Center of Sun Yat-Sen University from January 1999 to December 2009 were retrospectively analyzed. Of all the patients, 79received PRFA (PRFA group) and 72 received repeat hepatectomy (repeat hepatectomy group). The survival rate, morbidity and recurrence of the tumor between the two groups were compared. All data were analyzed using t test, chi-square test or Log-rank test, and the survival of the patients were analyzed using the Kaplan-Meier method. Results The mobidities of the PRFA group and repeat hepatectomy group were 13% (10/79) and 36%(26/72), respectively, with a significant difference between the two groups (x2=11.411, P＜0.05). The cumulative 1-, 2-, 3-, 4-, 5-year survival rates were 89.7%, 75.2%, 67.1%, 61.5%, 56.6% in the PRFA group, and 86.0%, 67.6%, 53.6%, 44.1%, 40.2% in the repeat hepatectomy group, with no significant difference between the two groups (x2=1.610, P＞0.05). The cumulative 4-, 5-year survival rates of the PRFA group were significant higher than those in the repeat hepatectomy group (x2=4.682, 4. 196, P ＜ 0.05). The local tumor recurrence rate of the PRFA group was 5% (4/79), and the incisal margin recurrence rate was 3% (2/72) in the repeat hepatectomy group, with no significant difference between the two groups (x2=0.565, P＞0.05). Conclusion As a less invasive treatment method, PRFA is superior to repeat hepatectomy for solitary recurrent HCC with the diameter≤3 cm.
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Objective To summarize the experience of adjuvant therapy for primary hepatocellular carcinoma. Methods 316 cases of operable hepatocellular carcinoma were divided into three groups. Only hepatectomy were performed in group one (21 8cases).Preopemtive adjuvant TACE were done in group two (52 cases). Preoperative adjuvant TACE and postoperative trans-portal vein chemotherapy were done in group three (46 cases), which was named hepatectomy sequencing two vessel therapy. Results 1, 3 and 5 year survival rote were 51.2 %, 30.0 % and 20.5 % respectively in group one, 57.2 %, 43.0 % and 31.5 % in group two, 84.0 %, 62.5 % and 51.0 % in group three. The postoperative disease-free survival rate in group three was significantly higher than that in group one and group two (P ＜0.05). Conclusion Hepatectomy sequencing two vessels therapy in perioperative period might improve the survival rate, which can prevent and delay the incidence of recurrence and may improve the effect of liver resection.
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Objective To construct replication deficient recombinant adenovirus of human monocyte chemoattractant protein-1(MCP-1) by homologous recombination.Methods The cDNA of MCP-1 gene was obtained from human liver tissue by using RT-PCR,and was subcloned into a transfer plasmid pAdTrack-CMV.The linearized recombinant transfer plasmid pAdTrack-CMV-MCP-1 was co-transformed with backbone vector pAdEasy-1 into bacteria BJ5183 for recombinant adenoviral plasmid.The recombinant adenoviral plasmid was linearized and then transfected into HEK293 packing cells to produce virus particles.The recombinant adenovirus was detected by using PCR.Results The recombinant adenoviral plasmid was successfully established and confirmed by restriction endonuclease digestion.The expression of green fluorescent protein(GFP) was observed on the 5th day after transfection.The fragment of MCP1 gene was amplified by PCR.Conclusion The achievement of recombinant adenoviral plasmid and recombinant adenovirus of MCP-1 lay a foundation for further investigation of the function and application of MCP-1.
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Objective To detect the expressions of p53, vascular endothelial growth factor (VEGF) and microvessel density (MVD) in breast cancer and tumor adjacent tissues to explore their relationship and roles in the pathogenesis of breat cancer. Methods The expression levels of p53,VEGF and MVD were assessed in 110 cases of breast cancer and tumor adjacent tissues with SP immunohistochemical method. Results The positive expression rate of VEGF and p53 was 90.0%(99/110) and 42.7%(47/110) in the breast cancer tissues, and 35.5%(39/110) and 4.5%(5/110) in the tumor adjacent tissues, respectively, which had highly significant difference between two groups (P
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Objective To establish a canine model of fulminate hepatic failure ( FHF) and study the treatment of FHF by partial orthotopic liver transplatation(POLT).Methods Carbon tetrachloride (CCL4) mixed with the same amount of peanut oil in the dosage of 0.9 ml per kilogram of body weight was injected intraperitoneally to canines. ALT,tolal bilirubine(TB), PT, NH 4,and blood suger(BG) were monitored. The pathological changes were observed when the canine died,remain alived animal were killed on 7th day and 14th day respectively. EEG was performed on 3rd day after the model eslablished. Results After CCl 4 was injected, the canine showed progressive hepatic failure, ALT, TB and NH rose persistantly, PT prolonged, and BG decreased (P