OBJECTIVE:To study the curative effect and mechanism of ulinastatin (UTI) administered by two different routes for severe acute pancreatitis (SAP). METHODS: 50 healthy male SD rats were randomly divided into 5 groups: sham operation (SO) group (fed with water ad libitum), SAP model group, positive control(nafamostat) group, ulinastatin peripheral venous perfusion group, and ulinastatin celiac arterial perfusion group. SAP model was established in the latter four groups, and then the rats were treated with corresponding drugs. 12h later, sample tissues were taken from pancreas and lungs for measurement of MPO level and histopathologic observation and the blood samples were collected for measurement of serum amylase, TNF-? and TXB2. RESULTS: Levels of amylase, TNF-?, TXB2, MPO and Hughes histopathologic scores of pancreas and lungs in all drug-treated groups were significantly lower than in SAP group (P

Objective To study the feasibility of using chitosan membrane to carry and transport melanocytes, in order to refine the technique for melanocyte transplantation with chitosan membrane. Methods Melanocytes were inoculated onto chitosan membrane and cultured for a period of time, then, electron microscopy,MTT assay and NaOH assay were carried out to estimate the adherence, growth and melanogenesis of the melanocytes. Skin wound surface was prepared in 12 nude mice, which were equally divided into 3 groups, test group inoculated with melanocytes on chitosan membrane, negative control group I treated with chitosan membrane without melanocytes, and negative control group II directly dressed immediately after the preparation of wound surface. On day 10 and 20 after the transplantation, confocal laser microscopy and immunohistochemistry were performed to observe the migration of melanocytes into the skin wound surface. Results Scanning electron microscopy and inverted microscopy showed that melanocytes were evenly distributed on and adhered well to the underlying chitosan membrane. As the growth curve of melanocytes demonstrated, chitosan membrane could support the normal growth of melanocytes, and no significant difference was observed in the synthesized melanin content between melanocytes cultured on the chitosan membrane and those in culture disks (0.087 ± 0.027 vs. 0.101 ± 0.036, t = 0.79, P > 0.05). Melanocytes were seen at the transplantation sites by confocal laser microscopy, and biopsy specimens from the transplantation sites stained positive for antimelan-A monoclonal antibody. Conclusions Melanocytes can adhere to and grow on the chitosan membrane,which can facilitate the migration of melanocytes to the transplantation sites in animals with the maintenance of biological activity of melanocytes.

Total caudate lobectomy via anterior hepatic transection is still a new technique to resect the tumor in the caudate lobe,which is mastered only by few surgeons.The procedure was successfully performed on a 21-year old patient with focal nodular hyperplasia in caudate lobe.The right and left lobes were first mobilized,then the short hepatic veins were dissected to detach the caudate lobe from the retrohepatic vena cava.Then the liver was split anteriorly and the partial middle lobe was resected.With this process,the tumor was in the sight and we dissected it from the liver parenchyma.The inflow blood was occluded 3 times with a period of 29,27 and 27 minutes,respectively,with an interval of 5 minutes.The total blood loss during operation was 1000 ml.The patient recovered quickly without any complications.The technique for caudate lobectomy via anterior hepatic transection can improve the success rate and safety of caudate lobectomy and deserve clinical consideration.

Retrograde caudate lobectomy is a proper technique to resect the tumor in caudate lobe when the tumor is too big or closely adherent to the inferior vena cava.A male patient aged 44 years was admitted to the Eastern Hepatobiliary Surgery Hospital in November 2007.The ligaments around the liver were firstly dissected to mobilize the whole liver,and the right hepatic pedicle was dissected and ligated,then the liver was splited anteriorly along the Cantlie's line.The tumor was opposed in the sight and then it was dissected from the liver parenchyma.The short hepatic veins were ligated and the tumor was detached from the inferior vena cava.The inflow blood was occluded for 19 minutes,and the total blood loss was 4500 ml.The technique of retrograde caudate lobectomy can improve the success rate and safety of caudate lobectomy when the tumor in the caudate lobe is too large or adherent to the inferior vena cava.

Objective To valuate different effects in hepatectomy of three inflow occlusion methods including pringle maneuver, hemihepatic occlusion and portal venous occlusion. Methods 180 patients undergoing hepatectomy were randomly assigned to pringle group (group A), hemehepatic occlusion group (group B)or PV occlusion group (group C). The amount of blood lost, measurements of liver enzymes alanine aminotransferase (ALT), aspirate aminotransferase (AST), total bilirubin (TB), Serum albumin (ALB) and postoperative complication were also recorded. Results There was no operative mortality. One patient in group B changed into pringle maneuver due to the difficulties in dissecting the hemi-hepatic portal and was excluded. The amount of hemorrhage of three groups had no statistical difference. The ALT, AST, ALB and TB level of 1,3,7 days after operation had significant differences in three groups. The pringle group had a higher level ALT,AST,TB and lower ALB level than the other two groups. Conclusions All techniques of occlusion are effective and feasible for patients undergoing hepatectomies. However, compared with pringle maneuver, PV clamping and hemihepatic occlusion can relieve the liver function damage after hepatectomy.

Objective To explore the safety and efficacy of hepatic resection with selective hepatic vascular exclusion(SHVE). Methods SHVE was used in 246 consecutive patients undergoing major or complex liver resection in our center. Preoperative demographic and clinical data,details of the surgical procedure, pathologic diagnosis, postoperative course and complications were collected and analy zed. Results From January 2000 to July 2007, liver resections were performed IJnder SHVE in 246 patients; total SHVE, right partial SHVE and left partial SHVE in 145, 54 and 47 patients, respectively. SHVE was converted to total hepatic vascular exclusion(THVE)in 3 patients to repair the wall of inferior vena cava(IVC). Hemodynamic tolerance to SHVE was excellent, only with a slight increase in systemic and pulmonary vascular resistance during clamping.There were no deaths. and the morbidity was 24.8%.The mean leyth of hospital stay was 9.6 days(range 8-18) .Conclusion Our study showed that SHVE is a safe and effective procedurei and it is applicable to liver tumors near but not invade the inferior vena cava.

The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Gene Expression ; Myocytes, Cardiac ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Transfection

Objective To study the clinical,imaging and pathological features of hepatic angiomyolipoma,and to investigate methods in improving the preoperative diagnosis rate.Methods The imaging features and treatment experience of 73 patients with hepatic angiomyolipoma who had been admitted to the Eastern Hepatobiliary Surgery Hospital from 2000 to 2007 were retrospectively analyzed.All patients were classified according to the imaging features and corresponding treatments were applied.Results Of all patients,7 were diagnosed preoperatively.The diagnostic rate of B ultrasound,computed tomography and magnetic resonance imaging were 0,13%(7/56)and 6% (2/33),respectively.According to the results of imaging examination,6 patients were with the type of hemangioma,17 with the type of lipoma,4 with the type of leiomyoma and 46 with mixed type.One patient was treated by radiofrequeney ablation and 72 by surgical resection.Twenty-four patients were presented with pulmonary infection,pleural effusion,ascites or slight hepatic dysfunction.Postoperative immunohistochemical assay demonstrated that HMB45 had the highest positive expression rate,then followed by smooth muscle actin,vimentin,proliferating cell nuclear antigen,CD34,polyclonal carcinoembryonic antigen,CD18,CD19 and p53.One patient died of postoperative tumor recurrence.Conclusions Hepatic angiomyolipoma is easy to be misdiagnosed,while imaging classification is helpful in the diagnosis.Surgical resection is beneficial to patients with hepatic angiomyolipoma.

Objective To investigate the significance of XAGE-1bmRNA expression in the blood specimens of patients with primary liver cancer, cirrhosis and benign liver diseases and in healthy individuals. Methods Venous blood specimens of patients with primary liver cancer (n= 125), cirrhosis (n= 23), benign liver diseases (n= 34) and healthy individuals (n = 41 ) were collected. XAGE-1b mRNA was detected by real-time fluorescence quantitative PCR. Results The expression levels of XAGE-1b mRNA in patients with primary liver cancer, cirrhosis, benign liver diseases and healthy in dividuals were 3.72 (0.93, 10.2) ×10-5, 0 (0, 0. 56) ×10-5, 0 (0, 0)×10-5, 0 (0, 0) ×10-5, respectively. The XAGE-1b mRNA expression in patients with primary liver cancer was obviously higher than the patients with cirrhosis, benign liver diseases and in healthy individuals. The expression levels for patients with cirrhosis was higher than patients with benign liver diseases and in healthy individuals. The expression levels for patients with benign liver diseases and healthy individuals were similar. With a optimal cut-off value of 8. 385 × 10-7 , the sensitivity, specificity, positive predictive value, and negative predictive value of XAGE-lb mRNA for diagnosing primary liver cancer were 80. 0%, 89.8%, 90.9% and 77.9% respectively. The positive rates for patients with primary liver cancer and cirrhosis were 80.0% and 30.4% respectively. Conclusion XAGE-lb mRNA can be used as a tumor marker for primary liver cancer. It contributes to the differentiation between primary liver cancer, cirrhosis and benign liver diseases.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.