High expression of DNMT3B promotes proliferation and invasion of hepatocellular carcinoma cells Hippo signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the role of DNMT3B in regulating the proliferation and invasion of hepatocellular carcinoma (HCC) cells.@*METHODS@#We collected the tumor tissues and adjacent tissues from a total of 175 patients with HCC diagnosed in the Second Affiliated Hospital of Chongqing Medical University between May, 2008 and May, 2013 to prepare the tissue microarrays. The association of the expression of DNMT3B with the prognosis and the tumor-free survival and tumor-specific survival rates of the patients was analyzed. Univariate and multivariate Cox regression analyses were used to analyze the effect of DNMT3B expression on the prognosis of HCC. We used RNA interference technique to knock down the expression of DNMT3B in Huh-7 hepatoma cells and observed the changes in cell proliferation using CCK-8 assay and EDU staining and in cell migration and invasion ability using Transwell assay.@*RESULTS@#The positive rates of DNMT3B was significantly higher in HCC tissues than in paired adjacent tissues (67.4% 41.1%, =0.015). A high DNMT3B expression in HCC was significantly associated with the tumor size (=0.001), vascular invasion (=0.004), and intrahepatic metastasis (=0.018). The patients with high DNMT3B expressions had significantly lower tumor-free and tumor-specific survival rates than those with low DNMT3B expressions ( < 0.005). In Huh-7 cells, silencing DNMT3B significantly inhibited the cell proliferation and inhibited cell migration and invasion. Western blotting showed that silencing DNMT3B obviously increased LATS1 expression, decreased the expression of YAP1, and activated Hippo signaling pathway. Methylation-specific PCR showed that the methylation level of LATS1 was decreased in the cells with DNMT3B silencing.@*CONCLUSIONS@#The expression level of DNMT3B is significantly higher HCC tissues than in the adjacent tissues, and the high expression of DNMT3B is closely related to the low survival rate of the patients. Silencing DNMT3B inhibits the proliferation, migration and invasion of HCC cells. DNMT3B promotes the progression of HCC primarily by enhancing the expression of YAP1 through methylation of LATS1 and inhibition of its expression, which inhibits the anti-cancer effect of Hippo signaling pathway.

The effect of HIFU combined with nanoscale ultrasound molecular probes with simple virus thymidine kinase gene on angiogenesis in the nude mouse tumor / 重庆医学

Objective To observe the change of the protein and gene expression of hypoxia inducing factor-1α(HIF-1a) and vascular endothelial growth factor (VEGF) in the nude mouse tumor,which has been treated by HIFU combined with nanoscale ultrasound molecular probes with HSV1-TK gene microvascular density.Methods Sixty nude mice were implanted with HepG2 Cells to establish subcutaneous transplanted tumor.Divided this mice into six groups at random after treated by HIFU:MB+ HSV-TK+ GPC3 (group A),MB + HSV-TK (group B),HSV-TK +GPC3 (group C),HSV-TK (group D),MB + GPC3 (group E),PBS (group F).They were injected into the tail vein every after 3 days.Mice in group A,B,D and E were exposed to ultrasound by 2 W/cm2,1 MHz,5 mintues and 0.2 mL ganciclovi(GCV) was intraperitoneally injected at the first 48 hours after injection.After the treatment,immunohistoche were used to detect the microvascular density(MVD),Western blot and immunohistoche was employed to test the protein change of the VEGF and HIF-1α,Q-PCR was used to test the mRNA gene transcription of VEGF and HIF-1α in the tumor tissues.Results After 14 days,the protein expression of HIF-1α and VEGF in group A was significantly lower than that in group B,C,D,E and F (P＜0.05),the MVD level in the tumor is also like this,and the difference is statistically significant.Conclusion Anoscale ultrasound molecular probes with HSV1-TK can reduce the the level of VEGF,MVD and HIF-1α in the tumor which has been treated by HIFU,so it can inhibit tumor growth and improve the therapeutic efficacy after HIFU treatment.

The experimental research on the protective effect of SD rats′liver transplantation reperfusion ischemia-reperfusion injury(IRI) after GW3965 activation of liver X receptor preprocessing / 国际检验医学杂志

Objective To study the protective effect and mechanism of SD rats′liver transplantation reperfusion ischemia‐reper‐fusion injury(IRI) after GW3965 activation of liver X receptor preprocessing .Methods Separated Male SD(Sprague‐Dawley) 70 rats into 3 groups which were sham operation group (SO group ,14 rats) ,orthotopic liver transplantation group (OLT group ,28 rats) ,and GW 3965 preprocessing group(GW 3965 group 28rats) .The levels of serum transaminase ,plasma inflammatory factors (TNF‐α、IL‐1) ,the changes of hepatic pathology and inflammatory factor mRNA ,and the activities as well as its expressions of NF‐κB in hepatic tissue were observed ,after the operation .Results After 6 and 24 hours perfusion ,the levels of plasma inflammatory factors was expression ,serum transaminase ,the liver pathological injury degree and the activities as well as its expressions of NF‐κB in OLT group and GW3965 group were higher than those in SO group .While after reperfusion for 6 and 24 hours ,the levels of ser‐um transaminase ,plasma inflammatory factors expression ,the liver pathological injury degree ,inflammatory factor and the activities as well as its expressions of NF‐κB in GW3965 group were much lower than those in OLT group ,there were obvious differences (P<0 .05) .Conclusion After GW3965 activation of liver X receptor preprocessing ,the activities of NF‐κB and the emerging of downstream inflammatory mediator factors are reduced effectively and protect the liver after the ischemia reperfusion .

Role of XBP1 signal transduction pathway in the ischemia/reperfusion injury of graft liver in rats / 中华器官移植杂志

Objective To explore the regulation mechanism of X box binding protein 1 (XBP1)signal transduction pathway for TNF-α and effective approach in ischemia/reperfusion (I/R) injury of liver transplantation for short hairpin RNA (shRNA) interference used to gene therapy in liver graft.Methods Male Sprague-Dawley rats were divided into three groups: the cold ischemia transfection group (CIT), the in vivo transfection group (IVT) and the control group. Experiments of orthotopic liver transplantation were performed by two cuff method. The rats in CIT were perfused with XBP1-shRNA plasmid (pSIXBP1) during cold ischemia phase, those in IVT received the equivalent volume (2 ml) of pSIIRAK 4 after portal vein inoculation, and those in the control group were not subjected to any treatment. Rats were killed at 60 or 180 min after restoring reperfusion of hepatic portal vein.Histopathological damage degree of graft liver was observed by light microscope. The expression levels of XBP1 gene and protein were detected by RT-PCR and Western blotting. The activities of NF-κB and the serum TNF-α level were detected by ELISA. Results All the indexes including the degree of histopathological damage, the expression levels of XBP1 mRNA and protein and the TNF-α level were significantly decreased in CIT as compared with IVT and control group (P＜0. 05). However,there was no significant difference in NF-κB activity among the three groups (P＞0. 05). Conclusion The role of XBP1 pathway in TNF-α gene regulation and that of NF-κB pathway in rat liver I/R injury are two relatively independent aspects, and the depression of XBP1 expression with XBP1 shRNA through portal vein perfusion during cold ischemia phase could effectively alleviate graft hepatic I/R

Current progress in bystander effect of suicide gene therapy / 国际外科学杂志

Nowadays, tumor is one of the most challenging human health diseases, which oblige human tirelessly to studied for the tumor.Gene therapy is a promising treatment in oncotherapies. Suicide gene therapy is the most widely researched among gene therapies. At the same time, bystander effect take important role in the mechanism of suicide gene therapy. Therefore, more researchers devote themselves to studyinghow enhance the bystander effect in order to improve the effect of suicide gene therapy. This article reviewed in short how to augment bystander effect of suicide gene therapy against cancer.

Experimental study on treatment of femoral head necrosis in rabbit with transplantation of bone marrow stromal cells / 中国矫形外科杂志

[Objective]To investigate the repairing effect and mechanism for treatment of femoral head necrosis on transplantation of bone marrow stromal cells.[Method]Twenty-four Newzland rabbits were divided into two groups randomly,Group A as core decompression group and Group B as Bone marrow stromal cells autograft group.Animal femoral head neorosis model was made by freezing method with liquid nitrogen.After freezing and drilling,The drilled necrotic cavities of Group A was implanted with gelatin sponge,while of Group B was with gelatin sponge combined by bone marrow stromal cells.Three animals in every group were sacrificed respectively at 2,4,6,8 weeks and subjected to radiograph and microscope examination.[Result](1)Radiograph examination:At 2 weeks,the drilled holes in Group A and B all presented low density images.At 4 weeks,the density in the drilled holes increased in group B and the density around the drilled holes increased in group A.At 8 weeks,sclerotic line formed around the drilled holes in Group A and trabeculae appeared in the drilled holes in group B.(2)Microscope examination:in group A,at 2 weeks,a few inflamatory cells appeared in the holes with many osteoblasts and ostoid appeared around the drilled holes.At 8 weeks,marrow formed in the drilled holes in group B,at 2 weeks,there were a large amount of osteoblasts in the drilled holes and some ostoid formed around thc drilled holes.At 4 weeks,the drilled holes were full of trabeculae.At 8 weeks,the trabeculae matured with the marrow appeared in the intertrabecular space.[Conclusion]Bone marrow stromal cells autografting may be an effective way to treat rabbit femoral head necrosis.

Application of problem-based learning pattern in teaching general surgery for probationers / 中华医学教育探索杂志

Problem-based learning(PBL) is a method of the self-study primarily and teacher' s guiding.By using the PBL in the clinical teaching application,we discuss our experience gained in carrying,out PBL and the existing problems.

Hemorrhage in laparoscopic cholecystectomy:the cause and management / 中国现代普通外科进展

Objective:To explore the cause and treatment of hemorrhage in laparoscopic cholecystectomy(LC).Methods:The clinical data of 112 cases of LC were analyzed to summarize the causes and treatment of hemorrhage.Results:The causes of hemorrhage in LC included subjective and objective elements.All of them were successfully hemostatic in different ways including reclamping,coagulation,suturing,packing hemostasis and suspension of falciform ligament of liver.Conclusion:Hemorrhage is the serious and most common complication in LC,but it can be avoided through an immediate and effective process.

Expression and influencing factors of CD54 in Hashimotos thyroiditis / 第三军医大学学报

Objective To dectect the expression of CD54 in thyroid tissue of Hashimotos thyroiditis patients,and expression changes of CD54 on thyroid cells interferred by different agents.Methods The thyroid tissues from 41 cases of Hashimotos thyroiditis were collected and 26 normal thyroid tissues served as normal controls.All thyroid tissues were identified by pathological examination.The positive expression rate and area of CD54 were investigated by immunohistochemical method and quantitative analysis of image analysis system in all thyroid tissues.The expression changes of CD54 in the isolated thyroid cells interferred by 100,500,1 000 pg/ml IL-1? or 10 mg/L NaI or 1 000 pg/ml IL-1? and 10 mg/L NaI for 48 h were detected by flow cytometry.Results The positive expression rate of CD54 in Hashimotos thyroiditis tissues was much more than that of control tissues(P

Expression of lipopolysaccharide binding protein and lipopolysaccharide receptor CD14 in experimental alcoholic liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the expression of lipopolysaccharide binding protein (LBP) and CD14 mRNA in alcohol-induced liver disease (ALD) and evaluate the relationship between the expression of LBP and CD14 mRNA and the severity of liver injury in alcoholic-fed rats.</p><p><b>METHODS</b>Twenty Wistar rats were divided into two groups: ethanol-fed group and control group. Ethanol-fed group were fed ethanol (by intragastric infusion of 500 ml/L ethanol orally, dose of 5~12 g/kg/d) and control group received dextrose instead of ethanol. Rats of both groups were sacrificed at 4 weeks and 8 weeks, respectively. Levels of endotoxin and alanine transaminase (ALT) in blood were measured, and liver pathology was observed by light and electronic microscopy. Expression of LBP and CD14 mRNA in liver tissues were determined with the reverse transcription polymerase chain reaction (RT-PCR) analysis.</p><p><b>RESULTS</b>Plasma endotoxin levels were increased significantly in ethanol-fed rats [(129 21) pg/ml and (187 35) pg/ml at 4 weeks and 8 weeks] than in control rats [(48 9) pg/ml and (53 11) pg/ml, respectively, t=11.2, 11.6, P<0.05]. Mean values for plasma ALT levels were increased dramatically in ethanol-fed rats after 4 weeks and 8 weeks [(112 15) U/L and (147 22) U/L, respectively] than in the control animals [(31 12)U/L and (33 9)U/L, respectively, t=5.9, 20.6, P<0.05]. In liver sections from ethanol-fed rats, there was marked pathological changes (steatosis, cell infiltration and necrosis). In the control rats, there was no significant difference in the levels of LBP and CD14 mRNA at the two time points. In ethanol-fed rats, ethanol administration led to a significant increase in LBP and CD14 mRNA levels as compared with the control group (P<0.05).</p><p><b>CONCLUSIONS</b>Ethanol administration lead to a significant increase in endotoxin levels of the serum and LBP and CD14 mRNA expression in liver tissues in ethanol- fed rats when compared with the control rats. Increase of LBP and CD14 mRNA expression may result in greater sensitivity to endotoxin and thus lead to liver injury.</p>

Effect of Peer -Education model on clinical practice of undergraduate interns / 中华医学教育探索杂志

Objective To investigate the effect of peer-education model on clinical practice of undergraduate interns.Methods Thirty-six undergraduate interns were arranged into control and experimental group.Twelve postgraduates were selected as peer-educators.Medical ethics and style,attitude,clinical thoughts,operations and satisfactory of patients were used to judge their work.They had to take exams at the end of the clinical practice.Results Undergraduate interns in experi-mental group got higher scores than control group.Conclusion Peer-education model has a posi-tive effect on clinical practice of undergraduate interns.

Tumor necrosis factor and endothelin and hyperdynamic circulatory syndrome in partial portal-vein ligated rats / 中国普通外科杂志

Objective To study the effect of tumor necrosis factor(TNF) and endothelin(ET) on the formation of murine hyperdynamic circulatory syndrome(HCS).Methods Circulation stable HCS model was produced in rats by partial portal vein ligation.Portal blood TNF and ET was determined pre and post injection of anti rat TNF and hemodynamics was monitored simultaneously.Results TNF level did not change 0 5 hour after partial portal vein ligation,but it increased significantly at 24 hour and maintained at this high level thereafter. ET was on a small increment at different time after partial portal vein ligation.The TNF level decreased significantly to the level in controls and ET remained unchanged after injection of anti rat TNF,while HCS was partially ameliorated. Conclusion [WT5”BZ] TNF may be related with the start and maintenance of HCS,and there is no causality between ET and the development of HCS.

Selection of anesthetic method in setting up the animal model of orthotopic liver transplantation in rats / 中国普通外科杂志

Objective To investigate the influences of anesthetic meltod on setting up the animal model of orthotopic liver transplantation(OLT) in rats, in order to properly select the anesthetic method. Methods OLT was performed by modified two-cuff technique in Sprague Dawley rats. 100 rats were randomly divided into 5 groups: control group, ether group, ketamine group, chloral hydrate group and pentobarbital group. The anesthetic process, the influences of anesthetic method on physiologic parameters and hepatic function, and the mortality during anhepatic phase of each group were observed. Results The anesthetic process of each anesthetic method was different,ether anesthesia had fewer impacts on physiologic parameters, and the pentobarbital had great hepatotoxicity. High mortality happened in ketamine group, chloral hydrate group and pentobarbital group during anhepatic phase, while no animal died in ether group. Conclusions Each anesthetic method has significant influences on the OLT rat. Ether anesthesia has fewer effects on physiologic parameters and liver function, and has low mortality during anhepatic phase of OLT rats.So it is the best anesthetic method in setting up the animal model of OLT in rats.

Effect of low dose irradiation on expression of membrane molecules of T lymphocytes from cord blood / 中国微创外科杂志

Objective To study the expression of membrane molecules of T lymphocytes of cord blood after low dose irradiation(LDI). Methods Freshly isolated lymphocytes from cord blood were irradiated with 62mGy gamma-ray. At different time(4h; 12h;24h) after irradiation, the changes of TCR+, CD3 +, CD4 +, CD8 + cells were examined by flow cytometry with direct immunofluorescence, respectively. Results The proportion of CD3 +, TCR+ /CD3 +; CD4 +, CD8 + cells increased significantly after LDI,the most obvious enhancement was noted in the 24h experimental group were no significant changes in the ratio of CD4 to CD8. Conclusions It is suggested that expedition of the maturation, activation and signal transudation of T lymphocytes from cord blood can be induced by ir- radiation with 62mGy gamma-ray The reconstruction of immune functions after cord blood transplantation may be ac- celerated, enhancing the graft versus leukemia(GVL)effect and preventing the tumor from relapsing.

Role of IRAK-4 activity in inhibitory effects of lipopolysaccharide pretreatment on hepatic ischemia/reperfusion injury / 中国病理生理杂志

AIM: To observe the changes of interleukin1 receptor associated kinase-4(IRAK-4) in ischemia/reperfusion(I/R) liver pretreated with lipopolysaccharide(LPS) and to explore the protective mechanisms of LPS pretreatment against hepatic I/R injury.METHODS: Male Sprague-Dawley rats,weighing 240-280 g,were divided into three groups: control,ischemia/reperfusion group(I/R group) and LPS-pretreated group(LPS group).On the first day,LPS group received 0.1 mg/kg LPS via the tail vein,followed by 0.5 mg/kg on the 2nd,3rd,4th and 5th day.I/R group received the equivalent volumes(0.5 mL) of sterile PBS.Experiments of I/R injury was induced by temporary ischemia of the left lateral liver lobe for 90 min followed by 3 h reperfusion on 2 days after the last LPS treatment.At 0 min,60 min and 180 min after reperfusion,the expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blotting.The activity of NF-?B and the serum TNF-? level were also detected by ELISA.RESULTS: Although the level of IRAK-4 gene and protein were higher in the LPS group than that in I/R group and control group(P0.05) at 0 min after reperfusion.However,all those indexes were evidently lower in the LPS group than those in I/R group(P

Change expressing of hTERT gene in breast cancer tissue before and after neo-adjuvant chemotherapy / 中国癌症杂志

Background and purpose:hTERT is indistinct in the effect of neo-adjuvant chemotherapy. Our aim was to detect both expression of hTERT mRNA and protein in breast cancer tissue and their changes after neoadjuvant chemotherapy. Methods:From Feb 2004 to Jun 2007, 53 cases with untreated advanced breast cancer were enrolled. All patients received CEF chemotherapy three cycles. The breast cancer tissue was collected before and after chemotherapy. hTERT mRNA and protein expression were detected by RT-PCR and immunohistochemistry techniques. Results:The positive hTERT mRNA and protein expression in breast cancer tissue before chemotherapy were 77.4% and 73.6% respectively, and after neo-adjuvant chemotherapy were 28.3%,22.6% respectively.The effective rate of the neo-adjuvant chemotherapy of the patients with positive of hTERT expression was higher than those with negative hTERT expression. Conclusion:The chemo-agents altered hTERT gene expression, improved negative rate of hTERT expression. Detecting expression of hTERT gene in breast cancer tissue would be an important index for predicting the effects of neo-adjuvant chemotherapy.

Effect of gadolinium chloride on suppressing acute rejection following liver allograft transplantation in the rats / 中华器官移植杂志

Objective To investigate the mechanism of gadolinium chloride (GdCl3) inhibiting Kupffer cells (KCs) to suppress acute rejection following liver allograft transplantation in the rats. Methods The rats were randomly divided into control group (A), liver transplantation with GdCl3 pretreatment group (B), and liver transplantation with normal saline pretreatment group (C). The survival rate, liver function, hepatic pathological histology, cytokine level in liver and bile, activity of NF-?B in KCs, and membraneous molecules on the KCs were observed. Results (1) The one-month survival rate in group B was significantly higher than that in group C (P

Determination of relative molecular weight in polysaccharide-2b of moutan(PSM_(2b)) / 中草药

Objective To determine the relative molecular weight(M_r) of polysaccharide-2b of moutan(PSM_(2b)) to establish the standard of reference substance.Methods PSM_(2b) was separated by a Sepharose 4B column and then PSM_(2b-A) and PSM_(2b-B) were obtained after cryodesiccation.Then HPLC was used to determine their M_r.Ultrapure water was used as mobile phase to compare with 0.7% Na_2SO_4 solution.Results PSM_(2b-A) was pure.The M_r and polydispersity(D) of PSM_(2b-A) obtained by use of dextran as standard were 3.596 2?10~(5) and 4.15 in 0.7% Na_2SO_4 solution phase,respectively,and in ultrapure water phase the M_r and D were 4.758 5?10~(5) and 1.03,respectively.Conclusion Different results are detected by different mobile phases in determination of M_r and D of PSM_(2b-A) by HPLC.And the ultrapure water as mobile phase is more suitable than 0.7% Na_2SO_4 solution.

Perioperative treatment of intra-and extra-hepatic cholelithinsis in patients with liver cirrhosis / 中国普通外科杂志

Objective To study the perioperative treatment of intra-and extra-hepatic cholelithinsis in patients with liver cirrhosis . Methods The clinical data of intra-and extra-hepatic cholelithinsis in patients with liver cirrhosis in our hospital in resent 10 years was retrospectively analyzed. Results According to the preoperative examation, improving hepatic function(turn child class C to A or B), correcting the coagulation disturbance,decreasing portal vein pressure preoperatively,and proforming operation carefully to reduce bleeding,and giving support treatment and liver care treatment to improve the liver function further postoperatively etc were made.Fifteen cases remained stones, 5 cases appearred chronic liver failure,2 cases appearred kidney failure ,the other 69 cases recovered well. Conclusions If optimizing perioperative treatment is given, favorable effect might be obtained in intra-and extra-hepatic cholelithinsis in patients with liver cirrhosis.

Construction of recombinant adenovirus vector expressing EGFP gene specifically in AFP producing liver cancer cells / 中国病理生理杂志

AIM: To construct a recombinant adenovirus vector carrying AFP promoter to specifically express a targeting gene in hepatocellular carcinoma HepG2 cells. METHODS: Based on the Adeno-X~TM Expression system, the CMV promoter was replaced by a 300 bp a-fetoprotein promoter. The EGFP(enhanced green fluorescent protein) gene as a report gene was inserted to the multiple-cloning site(MCS). The normal liver LO2 cells, hepatocellular carcinoma HepG2 cells and HeLa cells were infected by the recombinant adenovirus, respectively. Northern blotting and fluorescence microscope were used to detect the transcription level of EGFP gene and its protein expression, respectively. RESULTS: Northern blotting showed that the target gene was markedly transcribed in HepG2 cells, but slightly in LO2 and HeLa cells. Under the fluorescence microscope, strong EGFP expression was seen in HepG2 cells but very weakly in HeLa and LO2 cells. CONCLUSION: Under the control of the 300 bp human AFP promoter, the target gene carried by the recombinant adenovirus was expressed in the AFP-producing HepG2 cells at a very high level, but not or very weakly in AFP negative cells. This adenovirus system can be used as a new, potent and specific approach for the gene-targeting therapy for the AFP producing primary hepatoma. [