Objective:To investigate the correlations between CO-029 expression and cholangiocarcinoma invasion and metastasis, and the further explore the potential mechanism involved.Methods:The constructed lentiviral vector of vshRNA-CO-029 (LV/GFP/CO-029) was used to transfect and screen the stable transfected cholangiocarcinoma cell line HCCC-9810-vshRNA-CO-029 as the silence group, HCCC-9810 cells transfected with the mock plasmid were used as the mock group, and the untransfected cells were used as the control group. Cell scratch assay, Transwell assay and in vivo implantation assay were used to detect the migration, invasion and metastasis of the three groups of cells. Immunoprecipitation and tumor necrosis factor (TNF)-α stimulation assay were used to detect the effect of CO-029 on the expression of EMT-related genes.Results:The scratch healing rate of the silence group was (27.11±4.58)%, which was lower than that in mock group (92.84±6.24)%, the number of cells passing through Matrigel in silence group was (57.15±6.10), which was significantly lower than that in mock group (108.20±9.21) and control group (112.00±10.45), the differences were statistically significant ( all P<0.01). The volume of liver tumors in the silence group of orthotopic xenograft mouse model was (2.17±0.54) cm 3, while the volume of liver tumors in the transplanted simulation group was (0.74±0.15) cm 3, the differences were statistically significant ( P<0.05). The incidence of lung metastasis and the number of lung metastases in the simulated group was 100%(6/6) and (214.17±35.64), respectively, while that in the silence group was 16.7% (1/6) and (41.56±14.15), respectively, the differences were statistically significant (all P<0.05). Co-immunoprecipitation showed that CO-029 can form a complex with TNF-αR1. TNF-α induced the down-regulation of E-cadherin and up-regulation of vimentin and N-cadherin in the mock group, but no significant changes were observed in the silence group. Conclusion:CO-029 expression is positively correlated with tumor invasion and metastasis of cholangiocarcinoma, and could couple with TNF-α to induce EMT, which is a novel well-established potential prognostic and therapeutic target for cholangiocarcinoma metastasis and prognosis intervention.

Objective:To investigate the expressions and clinical significance of tetraspanin CO-029 and integrin αv in intrahepatic cholangiocarcinoma (ICC ).Methods:Tissue microarray (TMA) was used to detect the expression of CO-029 and αv in 254 cases of intrahepatic cholangiocarcinoma. The relationship between the two factors and clinicopathological features, recurrence, metastasis and prognosis was analyzed.Spearman method was used to analyze their correlation.Relationship between αv and CO-029 was studied by mass spectrometry and database search,immunoprecipitation and Western blot were used to detect the coexistence.Results:Tissue microarray analysis showed that the positive expression rate of CO-029 was 51.6% (131/254), and the positive expression rate of αv was 61.4% (156/254). The expression of CO-029 and αv were closely correlated with tumor envelope, size, number and TNM stage ( P<0.05). According to the time of recurrence (TTR), the expressions of CO-029 and αv in early postoperative recurrence group (TTR <1 year) were significantly higher than those in non recurrence group (TTR ≥ 1 year). The patients with high CO-029 expression were more likely to relapse ( HR=2.01, 95% CI=1.45-2.79; P<0.001) and had shorter survival time ( HR=2.03, 95% CI=1.46-2.81; P<0.001). The patients with high expression of αv had shorter recurrence time ( HR=1.85, 95% CI=1.38-2.47; P<0.001) and shorter survival time ( HR=1.95, 95% CI=1.40-2.71; P<0.001). Co immunoprecipitation and Western blot confirmed that αv and CO-029 formed a complex. There was a positive correlation between CO-029 and αv in intrahepatic cholangiocarcinoma ( r=0.401, P<0.01). Conclusions:The differential expression of CO-029 and αv were closely related to the recurrence, metastasis and prognosis of intrahepatic cholangiocarcinoma, and CO-029 may couple with αv to form a complex to promote the invasion and metastasis of intrahepatic cholangiocarcinoma.

Objective To investigate the effect of occult HBV infection (OBI) on carcinogenesis of cryptogenic hepatocellular carcinoma.Methods Samples of hepatocellular carcinoma (HCC) and pericarcinomatous tissues obtained after hepatectomy from January 2011 to November 2013 at the Third Affiliated Hospital of Sun Yat-Sen University were collected.They were divided into two groups:the cryptogenic HCC group (the CH group,n =26) and the HBV related HCC group (the HH group,n =40).These samples were compared with the normal liver tissues obtained in 30 patients.HBV DNA was identified by the nested polymerase chain reaction and the immunohistochemical method was taken to examine the hepatitis B virus X protein (HBx) and Yes-associated protein (YAP) expressions.Results OBI was identified in 20 (77.8％) cryptogenic HCC patients and 8 (26.7％) in the control group.There was a significant difference between the two groups (x2 =14.072,P ＜ 0.05).HBV DNA was detected in all the HBV-related HCC patients.The HBx protein expression was mainly located in the cytoplasm of liver cells and liver cancer cells,but YAP was expressed in the nucleus.Both of them showed diffuse brown or tan particles.In the HH group and the CH group,the positive expression rates of HBx protein in the tumorous tissues were 80.0％ and 90.0％,respectively,and 85.0％ and 82.5％ in the nontumorous tissues,but only in 40.0％ in the control group.The positive expression rates of YAP in the tumorous tissues were 65.0％ and 67.5％,respectively,15.0％ and 20.0％ in the nontumorous tissues,respectively,but only in 12.5％ in the control group.The HBx expression in the cancerous tissues and para-cancerous tissues of the HH group and the CH group showed no significant difference (P ＞ 0.05),but the YAP expression in the tumor tissues was significantly higher than that in the nontumorous tissues (P ＜ 0.05).The HBx and YAP expressions in the HH group were comparable to the CH group (P ＞ 0.05).However,their expressions in the cancerous tissue of the HH group and the CH group were significantly higher than in the normal liver tissues (P ＜ 0.05).Conclusion A high prevalence of HBV infection was observed in HBsAg-negative HCC and the high expressions of HBx and YAP might be involved in the process of cryptogenic hepatocellular carcinoma.

Objective To investigate the portal vein embolization (PVE) and portal vein ligation (PVL) in liver regeneration of rats with hepatic fibrosis.Methods Fifty rats with liver fibrosis were prepared,including 10 rats were randomly chosen as pre-operative control group.The other 40 rats were divided into two groups:PVE group (A1,n =20) and PVL group (A2,n =20).We chose to embolize and ligate the right portal vein,respectively.The blood samples were obtained at different end points for measuring ALT and AST levels.Each liver lobes and whole liver were weighed,and non-embolized liver lobe/whole liver weight ratio,non-ligated liver lobe/whole liver weight were caculated at different end points.The samples from liver with/without embolization or ligation were were stained with hematoxylin-eosin,and the changes of microstructure of liver were observed.Immunostained for PCNA and Ki-67 were performed.Results Transient elevation of postoperative ALT and AST levels were noted in each group.Serum ALT and AST reached the peak on the first day in both of PVE and PVL groups [ALT,A1 (66.5 ±6.3) U/L vs(491.5 ± 48.0) U/L,A2 group(62.8 ±5.7) U/L vs(433.7 ±41.0) U/L;AST,A1group (113.4 ± 12.5) U/L vs (685.2 ±65.7) U/L,A2 group (110.4 ± 11.1) U/L vs(623.9 ±75.2) U/L,P＜0.05),and started to decrease on the third day,recovered to the pre-operative level on the fourteenth day (P ＞ 0.05).The weight percentage of non-embolized and non-ligated liver lobes/whole liver after PVE and PVL increased.There was no significant difference between the first day and pre-operative levels (P ＞ 0.05).Nevertheless,there were significant differences observed from the third,seventh,fourteenth days (A1 group,50.2 ± 5.0,57.7 ±5.7,61.8 ±6.6;A2group,49.6 ±3.5,55.7 ±6.9,63.0±5.1,respectively)compared with preoperative groups (A1 group,34.4 ± 4.0;A2 group,34.4 ± 4.0) (P ＜ 0.05).There was no significant difference between group A1 and A2 in each time point (P ＞0.05).The PCNA and Ki-67 were positive in hepatocytes and increased after operation,reached the peak in the third day (P ＜ 0.05),decreased slowly and restored to the normal level in the fourteenth day after operation,meanwhile,there was no significant difference between group A1 and A2 (P ＞ 0.05).Conclusions Fibrosis rats had the ability of regeneration in the contralateral part of the liver after selective PVE and PVL and there was no significant difference on the proliferative degree.Therefore,the safety and reliability of PVE and PVL in inducing liver regeneration in rats with liver fibrosis were confirmed.

Objective To separate the CD133 + subpopulation in human hepatocellular carcinoma (HCC) and investigate the tumorigenicity.Methods The human liver cancer tissues were subcutaneously transplanted into nude mice to generate xenograft tumors which were then isolated to prepare single cell suspension.The expression of CD133 + subpopulation was further detected using flow cytometry.The CD133 + subpopulations were separated and depurated with magnetic-activated cell sorting system.Immunofluorescence was performed to identify the histological phenotype of CD133 + subpopulation.The in vitro and in vivo clone formation assay and in vivo xenograft formation assay were performed, respectively.Results Flow cytometry analysis revealed that a percentage of (4.1 ± 0.6) ％ CD133 + cells were detected in xenografts.Immunofluorescence studies showed that (86.8 ± 7.5) ％ of the isolated cells were CD133 +.Compared with CD133-population, CD133 + cells showed a higher capability to generate clone sphere in vitro and a higher tumorigenicity in nude mice (P ＜ 0.05).Conclusion The CD133 + subpopulation in human hepatocellular carcinoma had a potent tumorigenicity and was enriched in cancer stem cells.

Objective To study the effect of Tspan 8 on metastasis and invasion of human hepatocellular carcinomas(HCC).Methods RT-PCR and western blot were used to detect the expressions of Tspan 8 in HCC cell lines,HCC and matched nontumorous tissues.The expression of Tspan 8 was then down-regulated by LV/GFP/Tspan 8 in HCC cells.The expressions of Tspan 8 mRNA and protein were determined by RT-PCR and Western blot assay,respectively.The proliferation was examined by MTT,the expression of AMDM12 was assessed by Western blot,and the invasion ability of HCC cells was evaluated by transwells.Results A high level of Tspan 8 was found in high metastatic potential HCC cells,and the expression of Tspan 8 in HCC tissues was much higher than that in the matched nontumorous tissues. Down-regulation of Tspan 8 had no influence on the proliferation of HCC cells (P＞0.05),while it inhibited the expression of ADAM12 and the invasive ability of HCC cells (P＜0.01,P＜0.01 respectively).Conclusion Tspan 8 played an important role in invasion and metastasis of human hepatocellular carcinomas and down-regulation by LV/GFP/Tspan 8 inhibited the invasiveness of human hepatocellular carcinoma cells.

Enhanced extracorporeal counterpulsation (EECP) information consists of both text and hemodynamic waveform data. At present EECP text information has been successfully managed through Web browser, while the management and sharing of hemodynamic waveform data through Internet has not been solved yet. In order to manage EECP information completely, based on the in-depth analysis of EECP hemodynamic waveform file of digital imaging and communications in medicine (DICOM) format and its disadvantages in Internet sharing, we proposed the use of the extensible markup language (XML), which is currently the Internet popular data exchange standard, as the storage specification for the sharing of EECP waveform data. Then we designed a web-based sharing system of EECP hemodynamic waveform data via ASP. NET 2.0 platform. Meanwhile, we specifically introduced the four main system function modules and their implement methods, including DICOM to XML conversion module, EECP waveform data management module, retrieval and display of EECP waveform module and the security mechanism of the system.
Computer Communication Networks ; standards ; Counterpulsation ; methods ; Heart-Assist Devices ; Hemodynamics ; Humans ; Internet ; Medical Records Systems, Computerized ; standards ; Programming Languages

The development of external counterpulsation (ECP) local area network system and extensible markup language (XML)-based remote ECP medical information system conformable to digital imaging and communications in medicine (DICOM) standard has been improving the digital interchangeablity and sharability of ECP data. However, the therapy process of ECP is a continuous and longtime supervision which builds a mass of waveform data. In order to reduce the storage space and improve the transmission efficiency, the waveform data with the normative format of ECP data files have to be compressed. In this article, we introduced the compression arithmetic of template matching and improved quick fitting of linear approximation distance thresholding (LADT) in combimation with the characters of enhanced external counterpulsation (EECP) waveform signal. The DICOM standard is used as the storage and transmission standard to make our system compatible with hospital information system. According to the rules of transfer syntaxes, we defined private transfer syntax for one-dimensional compressed waveform data and stored EECP data into a DICOM file. Testing result indicates that the compressed and normative data can be correctly transmitted and displayed between EECP workstations in our EECP laboratory.
Computer Communication Networks ; standards ; Counterpulsation ; methods ; Humans ; Information Storage and Retrieval ; Medical Records Systems, Computerized ; standards ; Programming Languages

Objective To summarize the experience in the diagnosis and treatment of hepatic trauma. Methods The clinical data of 260 patients with hepatic trauma admitted from January 1988 to December 2007 were retrospectively reviewed with regard to degree of trauma, treatment methods, therapeutical effects, complications and SO on. Results One hundred and fifty-three eases were treated by operative management,1 07 cases by nonoperative management.236 cases were cured,24 cases died,and the case fatality rate was 10.2%.There were no death among 139 patients with hepatic trauma grades Ⅰ～Ⅱ,22 death among 119 grades Ⅲ～Ⅴ patients, all death of 2 in grade VI, which demonstrated the correlation between death and hepatic trauma grade was statistically significant. Complications appeared in 82 eases, mainly including Secondary hemorrhage, abdominal infection and so on. Conclusions Nonoperative management is suitable for hepatic trauma with stable hemodynamics. Operative management is rapidly selected when the hemodynamics aren't stable. The cooperation of many specialities can enlarge the application of nonoperative management and decrease complications.

Objective To explore the effect of long-term enhanced external counterpulsation(EECP)on endothelium-dependent and endothelium-independent vasorelaxation in the carotid arteries of atherosclerotic piss. Method Totally 18 20-day-old male infant pigs were randomly divided into 3 groups according to feeding given: the normal[control group(n=6),the hypercholesterolemic control group(n=6)and the hypereholesterolemic +EECP group(n=6).Porcine model of hypercholesterolemia was made by feeding high-cholesterol diet.After EECP for 36 hours in the hypercholesterolemic+EECP group(n=6),carotid arterial rings were harvested from all animals and their vaso-relaxation response to different dose of Acetylchofine(Ach)and Sodium nitroprusside (SNP)were detected,respectively.Results As the dose of Ach varying between 10-8 mol/L and 10-5mol/L, endothelium-dependent vasorelaxation ratio of hypereholesterolemic piss with or without EECP treatment was significantly lower than that of the normal control group(P＜0.05),however,endothehum-dependent vasorelax- ation ratio in pigs with EECP treatment was obviously higher compared with hypereholesterolemic pigs without EECP treatment(P＜0.05)as the Ach ranged from 10-7 mol/L to 10-5mol/L.Similarly,as the concentration of SNP ranged fiun 10-8 mol/L to 10-5 mol/L.endothelium-independent vasorelaxafion ratio of both the hypercholesterolemic control group and the hypercholesterolemic+EECP group were significantly lower than that of the normal control group(P＜0.05),and end othelium-independent vasorelaxation ratio of the hypercholesterolemic+EECP group was significantly higher than that of the hypercholesterolemic control group (P＜0.05).Condusions Long-term EECP improves the impaired endothelium-dependent and endothelium independent vasorelaxalion function resulting from atherosclerosis.