ObjectiveTo study the clinical manifestations, pathologic characteristics, imaging features, diagnosis and treatment of adenomas of extrahepatic bile duct.MethodsTwo cases of adenomas of extrahepatic bile duct in our hospital and 14 cases reported in the literatures were analyzed retrospectively.ResultsThe patients’(male 5, female 11) mean age was 58.4 years (range 21-85). The main manifestations included jaundice (n=11), abdominal pain (n=8),fever (n=6),dyspepsia (n=4),body weight loss (n=3) and claycolored stool (n=1). The locations of tumors were in the left hepatic duct (n=1), right hepatic duct (n=3), hepatic common bile duct(n=3）,the junction of cystic duct and common bile duct (n=1），distal common bile duct (n=8). The pathologic types were tubular adenomas (n=5), papillary (villous) adenomas (n=10),and mucous adenoma (n=1). All the patients underwent surgical therapy. The tumors were identified by postoperative histopathologic examination.ConclusionIt is difficult to correctly diagnose adenomas of extrahepatic bile duct before operation, because the clinical manifestations are usually atypical. The definite diagnosis should depend on histopathologic examination. It is the key to completely resect the tumors. Postoperative followup should be done on regular basis.
Objective To explore the feasibility of recombinant adeno-associated virus (rAAV) as a vector for the gene therapy of liver cancer. Methods The rAAV/enhance green fluorescein protein (EGFP) recombinant was prepared by the routine method of two plasmids cotransfection.Results The experiment showed that one 10cm plate could produce 107-108 infection unit recombinant by the method of two plasmids cotransfection, and the transduction of HepG2 cell was increased with the increase of infection dosage of rAAV. About 100 multiplicity of infection (MOI) AAV vector could make all the tumor cell light. Conclusion Liver cancer cell can be efficiently transduced by rAAV, and AAV vector may be a valuable vector for the gene therapy of liver cancer.
Objective To investigate the reversal of the multidrug resistant gene mdr1 in vivo by antisense oligodeoxynucleotide （ASODN） on the basis of study in vitro. Methods The cultured drug-resistant human hepatocellular carcinoma cells were injected under the skin of axilla to establish the tumor model of nude mice. mdr1 ASODN accompanied by Lipofectamine were injected locally and ADM was injected intraperitoneally. Control 1 and control 2 were locally injected by Lipofectamine and normal saline separately, and ADM was also injected intraperitoneally. Results As time went on the tumor size increased and from the 5th day on alterations were marked, tumor size in different time phase showed marked difference to the prior time phase with significant difference (P＜0.05）. Tumor size in group ASODN was marked smaller than that of other 3 groups after the 5th day (P＜0.05)，while tumor size of group control 1,2 and group SODN in different phase showed no significant difference (Pgt;0.05). The results suggested that SODN and Lipofectamine showed no marked effect on tumor growth of nude mice and ASODN had marked inhibition effect on tumor growth. Conclusion mdr1 ASODN can also reverse multidrug resistance of drug-resistant human hepatocellular carcinoma cells in vivo. After the treatment the tumor’s growth in nude mice will slow down in a range of time.
ObjectiveTo review the causes, prevention methods, and therapies of the small-for-size syndrome (SFSS) in living donor liver transplantation (LDLT). MethodsThe literatures about SFSS in recent years were reviewed and summarized. ResultsThe donor’s age, graft steatosis level, MELD score of the recipient, portal hypertension, low outflow, and graft size were risk factors of SFSS. Ideal donor, splenectomy, ligating splenic artery, keeping a satisfactory intraoperative outflow, early diagnosis and active therapy could obviously decrease the incidence of SFSS. ConclusionThe risk factors of SFSS can be predicted before operation, and the positive actions can be used to prevent or cure the SFSS.
Objective To approach the prognosis after liver transplantation （LT） of liver function for Child grade A in patients with portal hypertension， and to compare with periesophagogastric devascularization with splenectomy （PDS）. Methods The data of 195 portal hypertension cases with Child A caused by hepatitis B cirrhosis who received surgical treatment of PDS （152 cases） or LT （43 cases） in division of liver transplantation center of West China Hospital of Sichuan University from 1999 to 2011 were retrospectively analyzed. The pre-， intra-， and postoperative variables in two groups that including patients’ age， score of Child， score of model for end-stage liver disease （MELD）， total bilirubin （TB），creatinine （Cr）， international normalized ratio （INR）， albumin （Alb）， complications of portal hypertension， amount of intraoperative bleeding and blood transfusion， operative time， and in the ICU and hospital stay time were compared. The postoperative outcomes were statistically analyzed including severe postoperative complications， short-term and long-term survival rates. Results Compared with PDS group， the amount of intraoperative bleeding and blood transfusion of LT group were morer （P＜0.05）， the operative time， in the ICU and hospital stay time of LT group were longer （P＜0.05）. The rate of severe postoperative complications in LT group was higher than that in PDS group 〔18.60% （8/43） vs. 1.97% （3/152），P＜0.05〕. The levels of TB and Cr during the postoperative period in LT group were higher than that in PDS group （P＜0.05）. Although the INR on day 1 after operation in LT group was higher than that in PDS group （P＜0.01）， but the difference disappeared soon on day 7 after operation in two groups （P＞0.05）.The 1-， 3-， and 5-year survival rates of the LT and PDS groups were 90.3%， 86.5%， 86.5%， and 100%， 100%， 100%， respectively， significant difference were observed in both short-term and long-term survival rates between the two groups （P＜0.05）. Conclusion LT offered no significant survival benefit to patients with portal hypertension and Child A due to hepatitis B cirrhosis， whereas PDS could be an effective treatment.
Objective To study the expression and significance of multidrug resistance-associated protein (MRP) gene in hepatocellular carcinoma (HCC). Methods Reverse transcription polymerase chain reaction (RT-PCR) assay was used to detect the expression of MRP mRNA in 25 fresh specimens of the primary HCC and its surrounding liver tissues. Immunohistochemistry LSAB technique was adopted to test MRP in 60 HCC specimens. The drug sensitivity was also tested by flow cytometry.Results The positive expression rates of MRP mRNA and MRP protein in primary HCC were 44.00%(11/25) and 45.00%(27/60) respectively. All the intensity of expression was low, but significant higer than its surrouding liver tissues (P＜0.05). The intensity and expression rate of MRP protein in 5 recurrent HCC had a tendency to increase. There was a correlation between the expression of MRP mRNA and MRP protein in 25 patients using RT-PCR and immunohistochemistry assay (Plt;0.05). Detected by flow cytometry, the average sensitivity of drugs in vitro of 60 HCC sp-cimens were 5-FU （15.80±7.63）%，DDP（18.45±9.59）%，ADM（17.95±7.99）%，MMC（16.60±8.69）% and CTX（17.40±10.14）%. Only 5FU and ADM were significantly affected by the expression of MRP protein (Plt;0.05).Conclusion The expression of MRP in primary HCC may be one of the important mechanisms of the intrinsic and acquired drug resistance in HCC. To study the expression of MRP could give a predictive value in HCC chemotherapy.
Objective To summarize the experience of the prevention of early arterial compl ications after hepatic artery (HA) reconstruction in adult-to-adult l iving donor l iver transplantation (A-A LDLT). Methods Between January 2002and March 2008, 127 patients underwent A-A LDLT. Of the 131 donors (127 cases of right lobe graft, 4 cases of left lobe graft), there were 69 males and 62 females with a mean age of 36.2 years (range, 19-65 years); in 127 recipients, there were 109 males and 18 females with a mean age of 41.9 years (range, 18-64 years). All patients underwent microsurgical reconstruction of HA between grafts and recipients. The artery of graft was anastomosed to the right HA in 62 cases, to the proper HA in 34 cases, to the left HA in 7 cases, to the common HA in 6 cases, and aberrant right HA rising from superior mesenteric artery in 8 cases. Interposition bypass using great saphenous vein (GSV) was performed between the donor right HA and recipient common HA in 5 cases. Bypass was performed between the donor right HA and recipient abdominal aorta using GSV in 2 cases, or using cryopreserved cadaveric il iac vessels in 3 cases. Results Of these 127 cases, hepatic artery thrombosis (HAT) occurred in 2 recipients (1.6%) at 1 day and 7 days following A-A LDLT, which were successfully revascularized with GSV between right HA of donor and abdominal aorta of recipient, HAT in 1 patient occurred on the 46th postoperative day with no symptom. No other arterial compl ication such as HA stenosis and aneurysm occurred in recipients. All patients were followed up 9-67 months. At 1, 2, and 3 years, actual survival rateswere 82.2%, 64.7%, and 59.2%. No death was related to HA compl ication in peri-operative period. Conclusion The anatomic structure and variation of HA, the pathological changes, as well as surgical technique in HA reconstruction, have direct impact on the risk of postoperative compl ications of HA reconstruction.
【Abstract】ObjectiveTo construct an mdr1 expression vector and detect its expression in HepG2 cells in vitro. MethodsThe 4.5-kb mdr1 cDNA was obtained from the plasmid pHaMDR1 cloned into the PCIneo mammalian expression vector, which was later transferred into human hepatocarcinoma cell line HepG2 by liposome. Then the HepG2 cells resisting G418 were clustered and proliferated，and the specific fragment of mdr1 cDNA, mRNA and the Pgp in these HepG2 cells were detected by means of PCR, RT-PCR and FCM respectively. ResultsThe mdr1 expression vector was constructed successfully，and the stable multidrug resistance(MDR) hepatocarcinoma cell line （HepG2/mdr1） was developed as well. The outcome of PCR analysis showed that the specific fragment of mdr1 cDNA could be found in HepG2/mdr1 cells, but not in the nontransfection HepG2 cells. Furthermore,the content of the specific fragment of mdr1 mRNA and the expression of P-gp in HepG2/mdr1 cells were (59.7±7.9)% and (12.5±5.45)% respectively, the corresponding value in HepG2 cells were (16.9±3.2)% and (4.63±2.59)% respectively. The difference was statistically significant (P＜0.05). ConclusionIt is praticable to develop MDR hepatocarcinoma cell line by transferring mdr1 cDNA into HepG2 cells, which is useful in the research of MDR mechanism.
Objective To analyse the clinical features of Peutz-Jeghers syndrome and its canceration, and to summarize the management and the follow-up strategy for patients with Peutz-Jeghers syndrome. Methods Clinical data of 30 patients with Peutz-Jeghers syndrome dating from October 1985 to September 2005 in West China Hospital of Sichuan University were analyzed retrospectively. Results Fourteen (46.67%) definite family histories of Peutz-Jeghers syndrome were found. Pigmentation of skin and mucosa, abdominalgia and hematochezia were major clinical manifestations of the syndrome. There were 18 patients (60.00%) complicated with intussusception and acute intestinal obstructions, 16 patients (53.33%) with gastrointestinal bleeding, and 6 patients (20.00%) developed malignancy. The frequent types of malignancy were carcinoma of small intestine (3 cases), colon carcinoma (2 cases) and gastric carcinoma (1 case) in order. The mean age of the canceration-diagnosed patients was 32 years old. The type of pathohistology of all the malignancy was poorly differentiated mucus adenocarcinoma. High-frequency endoscopic electroresection, orthdox polypectomy and enterectomy were the major means of treatment. Conclusion Patients with Peutz-Jeghers syndrome are at high risk of canceration at relatively early ages and usually the differentiation of the tumor is poor. Endoscopy should be performed regularly and the high-frequency electroresection is an effective therapy in disposing intestinal polyp. Screening can also improve the efficacy of Peutz-Jeghers syndrome.
【Abstract】Objective To construct a recombinant adenoviral vector carrying antisense matrix metalloproteinase2 (MMP2) for use in the gene therapy to inhibit the invasiveness and migratory capacity of hepatocellular carcinoma (HCC) cell line HepG2 in vitro and in vivo models. Methods Total RNA was extracted from HCC, and then a 500 bp fragment at the 5′ end of human MMP2 cDNA was synthesized by polymerase chain reaction (PCR) and was reversely inserted into the multiclone site (MCS) of the shuttle plasmid pAdTrack-CMV,with the resultant plasmid and the backbone plasmid pAdEasy-1,the homologous recombination took place in the E.coli BJ5183 and the recombinant adenoviral plasmid carrying the antisense MMP2 gene was constructed and generated. The adenoviruses(Ad-MMP2AS) were packaged and amplified in the HEK 293 cells.Then the viral titer was checked by GFP. Results The recombinant adenovirus vector carrying antisense MMP2 was constructed successfully, the b green fluorescence was observed in HEK 293 cells under a fluorescence microscopy. The viral titer was 1×108/ml. Conclusion The recombinant adenovirus Ad-MMP2AS constructed by us could introduce the antisense MMP2 into HepG2 effectively,which would provide experimental basis for reversing the overexpression of MMP2 in HCC and for inhibiting the invasiveness and migratory capacity of HepG2 in vitro and in vivo models.