ABSTRACT
The development of cholangiocarcinoma is slow, but its metastasis is extremely quick.When the tumor is found, the patients often have missed optimal operation period, and there is a lack of effective diagnostic indexes at present.In recent years, with the rapid development of molecular biology, studies find that microRNAs (miRNAs) are closely associated with cholangiocarcinoma.Because of miRNAs have great stability in various types of body fluids, so miRNAs in serum/plasma and bile are expected to become potential biomarkers in the diagnosis of cholangiocarcinoma.
ABSTRACT
Objective To establish a method of inducing dendritic cells(DC)from rat bone marrow cells in vitro,and identify the phenotype and function characteristics.Methods The rat bone malToW cells were collected and cultured in vitro under the condition of recombinant rat GM-CSF(rrGM-CSF)and recombinant rat IL-4(rrIL-4).After 2 weeks,the morphological character of DCs was observed under light microscope and scanning electron microscope.Expression of MHC-Ⅱ,CD80 and CD86 were detected by flow cytometry.The ability to stimulate allogenic T cells of the cultured DCs was detected by mixed lymphocyte reaction.Results DCs showed typical morphology with elongated dendritic processes under inversion microscope and scanning electron microscope.DCs at day 6 revealed immature phenotype,including MHC-Ⅱ(29.03 ±4.39)%,CD80(21.98±7.08)%and CD86(25.94±6.80)%.DCs at day 12 showed higher expression of MHC-Ⅱ(74.05±5.97)%,CD80(79.85±6.53)%and CD86(81.00±7.47)%,and stimulatory capacity of allogenic T cells,compared with that in DCs at day 6.Conclusion Matured DCs could be generated from rat bone marrow cells and attendance with rrGM-CSF and rrIL-4,which present the feasibility for further research on its application to allograft immunorejection.
ABSTRACT
BACKGROUND: Kadsurenone is characterized by obvious anti-oxidation and calcium ion resistant effect and can effectively clear oxygen-derived free radicals, decrease activity of pancreatic enzyme, and relieve self-digestion of pancreatic tissue.OBJECTIVE: To observe the effects of kadsurenone on the splanchnic blood flow in rats with severe acute pancreatitis (SAP), especially the mechanism of pancreatic blood flow.DESIGN: Contrast observational animal study.SETTING:Laboratory of General Surgery and Central Laboratory of Tangdu Hospital, the Fourth Military Medical University of Chinese PLA.MATERIALS: A total of 40 male Sprague-Dawiey (SD) rats, of clean grade and weighing 200-280 g, were selected in this study. Kadsurenone (150 mg) was provided by Pharmacological College of Peking University (batch number: 20050347); sodium taurocholate by Sigma Company (batch number: 86339); amylase reagent by Roche Company (batch number: irp 170505); Imexlab9100 Doppler ultrasound diagnostic meter by Imex Company; 8 mHz probe privated by Doppler ultrasound diagnostic meter by Imex Company, USA (number: X1-M0008).METHODS: The experiment was carried out in the Laboratory of General Surgery, Tangdu Hospital, Fourth Military Medical University of Chinese PLA from April to October 2006. Eight rats were randomly selected as sham operation group, and other rats were used to establish SAP models at 12 hours after fasting. Continuously, 32 SAP rats were randomly divided into two subgroups after model establishment, including kadsurenone group (n =16) and model group (n =16). In addition, all rats in the two subgroups were observed at 12 and 24 hours after operation with 8 rats at each time point. However, rats in the sham operation group did not give any treatment. At 3 hours after operation, rats in kadsurenone group were given 10 mg/kg kadsurenone through jejunum fistula once every 6 hours for one or two times in total. Kadsurenone was dissolved in 25 g/L dimethyl sulfoxide. In addition, rats in model group were given the same dosage of dimethyl sulfoxide through jejunum fistula; furthermore, rats in sham operation group were only opened their abdomens. Blood starch was respectively measured in sham operation group at 12 hours after abdominal operation, model group at 12 hours and kadsurenone group at 24 hours after acute pancreatic operation; meanwhile, Doppler ultrasound was used to detect local artery of pancreas (relative to the crossed site between head and body of pancreas), blood flow of portal vein, splenic artery and blood flow of superior mesenteric artery.MAIN OUTCOME MEASURES: Changes of plasma amylase and blood flow at various time points.RESULTS : All 40 SD rats were involved in the final analysis.①Measuring results of plasma amylase: Level of plasma amylase was increased in both model group and kadsurenone group at various time points as compared with that in sham operation group (P＜0.01 ). With the time passing by, level of plasma amylase was still increased in model group, and there was significant difference from that at 12 hours after operation (P＜0.05). ②Blood flow of local pancreatic artery: At 12 and 24 hours after operation, blood flow was lower in kadsurenone group and model group than that in sham operation group (P＜0.01). With the development of illness course, blood flow was still decreased in kadsurenone group (P＜0.05); however, blood flow was obviously higher than that in model group at various time points (P＜0.01 ). ③ Blood flow of superior mesenteric arterial root, portal vein and splenic artery: Blood flow was lower in kadsurenone group and model group than that in sham operation group at 12 and 24 hours after operation, respectively (P＜0.01). In addition, blood flow was remarkably increased in kadsurenone group as compared with that in model group at various time points (P＜0.01).CONCLUSION: Kadsurenone can increase the pancreatic blood flow and relieve pathological injury in SAP rats.
ABSTRACT
Objective: To construct and identify the eukaryotic expression vector targeting the CD40 gene in rats.Methods: Two sequences corresponding to the rat CD40 gene were designed on Ambion's Web.The two complementary oligonucleotide strands of DNA fragments were synthesized by chemosynthesis.After annealing of the complementary strands,the DNA fragments were connected to the polyclone sites of the pSilencer 4.1-CMV neo vector,followed by transformation,amplification,plasmid extraction and identification of the recombinant plasmids by BamH Ⅰ and Hind Ⅲ digestion and DNA sequence analysis.Results: The connections between the DNA fragments encoding CD40-targeted siRNA and the pSilencer 4.1-CMV neo vector were correct,as confirmed by agarose gel electrophoresis and DNA sequence analysis.Conclusion: The two recombinant plasmids expressing siRNA of the rat CD40 gene were successfully constructed,which prepared the groundwork for future research on the role of the CD40 gene in homograft rejection.
ABSTRACT
Objective: To observe the expressions of NMDAR2 and GLAST in the hippocampal gyrus and supraoptic nucleus(SON) under sleep deprivation in young rats.Methods: The changes in expressions of NMDAR2 and GLAST in SON and hippocampal gyrus under sleep deprivation in younger rats were observed by immunohistochemistry method. Results:The expressions of NMDAR2 and GLAST in SON and the hippocampal gyrus were significantly increased on the 3rd day of sleep deprivation,even more sigificantly on the 5th day,decreased gradually on 7th day,and became similar to those of control group on the 14th day.Conclusion: Sleep deprivation can affect the expressions of NMDAR2 and GLAST in the hippocampal gyrus and supraoptic nucleus,but the effect diminisshes gradually with prolonged time,which may be associated with self-regulation and self-protection of the central nervous system.
ABSTRACT
Objective To evaluate the results and feasibility of US-guided radio-frequency(RF) ablation for the treatment of primary or secondary liver tumors.Method 180 patients with liver tumor were treated with percutaneous ultrasound-guided RF ablation.RF was applied by a frequency at 10W with an increase of 10W every other minute until 90W for a total of 15 minutes,using single or multi puncture method.After RF treatment,the α-fetoprotein(AFP) level was examined,the ultrasonograph,computered tomography(CT),biopsy were performed.Results In those with abnormal pretreatment AFP level,1 week after RF treatment AFP returned to normal in 119/153(77.8%)patients;2-4 weeks later or after the second treatment AFP drops to normal in 144/153(94.1%) patients.Tumors failed to grow or shrank in size,some underwent liquefaction on CT and ultrasound follow-up.The biopsy results of the treated lessions showed necrosis accompanied with microvessel thrombosis.Abdominal pain disappeared or relieved in 121/142(85.2%) patients.Ascites and temporary jaundice occured in 26/180(14.4%),and 16/180(8.9%) patients,respectively.Conclusion RF ablation is a mini-invasive,safe,effective treatment for primary and secondary liver tumor.
ABSTRACT
Objective To investigate the causes,diagnosis and treatment of postoperative acute cholecystitis. Methods Clinical data of 9 cases with postoperaive acute cholecystitis were analyzed retrospectively. Results7 cases was confirmed by ultrasonography, 1 case was confirmed by CT, and 1 case died of gallbladder perforation, no operative mortality in emergency cholecystectomy. Conclusion The prevalent etiology is biliary stasis. Early diagnosis and treatment is the key point to decrease the death rate.
ABSTRACT
To observe the effect of liver transplantation on gastric mucosa lesions and the changes in plasma superoxide dismutase(SOD),malondialdehyde(MDA),interleukin 1(IL 1) and prostaglandin(PG) between before and after liver transplantation, a hepatic cirrhosis model was established in rats by subcutaneous injection of 40% carbon tetrachloride. Rat liver transplantation was made with the two cuff technique. By immunohistochemical method,we observed the effect of liver transplantation on rat gastric mucosa lesions. Using chemical method the changes in plasma SOD, MDA,IL 1 and PG between before and after liver transplantation were observed.The results showed that gastric mucosal lesions alleviated significantly after liver transplantation in rats with liver cirrhosis,and lesion index decreased gradually.The plasma levels of SOD decreased,MDA increased,and the plasma levels of IL 1, PGE 2 increased gradually after liver transplantation. Therefore, liver transplantation can improve gastric mucosal lesion, and this effect may be connected with its action on oxygen radicals, PG and IL 1.
ABSTRACT
Objective To investigate and evaluate the changes of T lymphocytes and red cell immunity of peripheral blood in patients with primary hepatocyte carcinoma (PHCC) after radiofrequency ablation(RFA) treatment. Methods The pre- and post- RFA(3d,7d,14d) peripheral blood T lymphocyte subsets(T3,T4,T8,T4/T8) and red cell immunity (RBC C3 receptor flower and RBC-immuocomplex formation rate) were investigated in 120 patients with PHCC treated by RFA. Results On 7d, 14d after RFA, T3, T4 lymphocytes and T4/T8 were higher than those on preoperative day significantly(P
ABSTRACT
Objective To evaluate the efficacy of ultrasonically guided radiofrequency (RF) thermal ablation of hepatic carcinoma (HC) with a clustered electrode. Methods Clinical records on 245 consecutively admitted HC patients who underwent RF tissue ablation therapy were analyzed. The tumor supplying blood flow, changes of tumor size, clinical manifestation, and AFP level were analyzed. Results AFP value decreased significantly in 50% of patients one week after ablation. Tumors less than 5*!cm in diameter became avascularized 1 to 6 months after the therapy, and it was noted that all tumors shrank in size after the therapy. For tumors more than 5*!cm in diameter, RF ablation was used in conjunction with intraarterial chemotherapy and embolization. Conclusion Ultrasound guided RF thermal ablation is an effective, safe, minimally invasive technique for treating malignant hepatic tumors.
ABSTRACT
Objective To explore the changes of IL-6, IL-8 and TNF-? levels before and after pericardial(devascularization) associated with proximal spleno-renal shunt(combined operation) in cirrhotic patients with portal hypertension. Methods Plasma levels of IL-6, IL-8 and TNF-? in combined(operation ) patients at 1d before(operation),and 7d after operation were measured and compared with control group. Results Preoperatively, the(plasma) levels of IL-6[ (230.4?10.3) ng/L], IL-8 [ (201.5?7.8) ng/L]and TNF-?[ (261.5?28.3) ng/L]in(combined) operation patients were significantly higher than those in control group (P
ABSTRACT
Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.
ABSTRACT
Objective To study the death mechanism of LoVo cells transfected with retroviral vector carrying cytosine deaminase (CD) suicide gene(G1CEACDNa). Methods Plasmid G1CEACDNa was transferred into the LoVo cells using liposomes method. RT-PCR was performed to detect CD mRNA expression using the total RNA extracted with TRIzol reagent. After exposed to 5-FC (1mmol?L -1 ), the cell growth inhibition rate and the “bystander effect” were determined by MTT, the cell ultramicroscopic structure was observed by electron microscope, and cell apoptosis was detected by flow cytometry. Results RT-PCR detection showed that there was 1.5kb band of CD product. The growth of the trasfected LoVo cells started to be inhibited after exposed 5-FC for 48h, and the inhibition rates at 72h,96h and 120h were 30%,50% and 80%, respetively.Electron microscope examination showed that there were apoptotic body. Meanwhile,a few cells showed necrosis alteration.Flow cytometry analysis showed that a few cells appeared apoptosis after exposed to 5-FC for 48h, the apoptotic rate and the necrosis rate at 72h and 96h were 23.8%, 35.1% and 20.4%, 26.0%, respectively. Conclusion The death mechanism of LoVo cells transfected with plasmid G1CEACDNa followed 5-FC treatment was involved in both necrosis and apoptosis. Apoptosis possibly was a mechanism of the bystander effect.
ABSTRACT
To study the effect of chinese herbal medicine preventing gallstone formation, morphology of livers and gallbladders as well as conjugated bile acids of the bile. 58 female guinea pigs were randomly divided into three groups. The control group was fed on normal diet; the lithogenous groups was high cholesterol diet (HCD) and the antilithic group was high cholesterol and the chinese herbal medicine capsule (13.5 g/kg). 8 weeks later the guinea pigs were killed. Liver and gallbladder were removed for light and electroscope observation. Gallbladder bile were withdrawn and gallbladder were collected. The minute construction of gallstones were observed by scanning electron microscope and gallstone components were quantified by Fourier Transform Infrand Spectros Copy (FT-IR). The morphology of liver and gallbladder were observed by light microscope and transmission electron microscope. Conjugated bile acid of biles were determined by Thin-layer Chromatograph. The effect of chinese herbal medicine preventing gallstone formation, morphololgy of livers and gallbladders as well as conjugated bile acids of the bile were also observed. We measured conjugated bile acids in all groups and found that decreased in lithogenic group and increased notably in antilithic group (P＜0.01). The gallstone formation rate was 100% when the animals were fed on HCD in lithogenic group. The rate decreased to 5.3% when the animals were fed on HCD and Chinese herbal medicine capsule inantilithic group. It indicates that changes in diet is an important factor in the process of gallstone formation and the chinese herbal medicine can prevent gallstone formation effectively.
ABSTRACT
Objective To investigate the changes of the levels of TNF and IL 8 in blood and in gastric mucosa and their mRNA expression after liver ischemia reperfusion (I/R). Methods 130 rats were randomly divided into 3 groups: (1) sham operation group(control group, n=10);(2)liver ischemia reperfusion group (n=60), including ischemia for 20min and 40min, and reperfusion for 1h, 24h and 72h respectively; (3) liver I/R rats treated with famotidine group (n=60). ALT and AST were detected after operation, pathomorphological changes of gastric mucosa were studied by light microscope and transmission electronic microscope, the concentrations of TNF and IL 8 in blood and gastric mucosa were measured by radioimmunoassay (RIA), TNF mRNA and IL 8 mRNA were detected by in situ hybridization. Results After liver I/R,inflammatory damage appeared in gastric mucosa, TNF and IL 8 concertrations increased significantly(P
ABSTRACT
Objective To investigate the effect of thermochemotherapy with 5 fluorocytosine(5 FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue specific cytosine deaminase (CD) in vitro.Methods Recombinant retroviral vector G1CEACDNa, in which CD gene was controlled under the CEA promoter, was introduced through liposome technique to the human colorectal carcinoma cell line SW480, and then the cells were selectively cultured in G418. The proliferated colonies were treated with the combined therapy of 5 FC and hyperthermia at a 43℃ over 30 min for 3 times. RT PCR was performed to analyze the expression of CD gene in target cells after thermochemotherapy. Results The CD gene expressed steadily in the target cells after 3 times of hyperthermic treatment. After the transfection of CD gene, SW480 CEACD cells were more sensitive to 5 FC than their parental cells (P
ABSTRACT
Objective To investigate the diagnosis and treatment of cystic adenoid tumor of the pancreas. Methods Retrospective analysis was made on the clinical data of 11 cases of pancreatic cystic neoplasms confirmed by operation and pathology at our hospital from 1978 to 1998. Results Cystic adenoid tumor of the pancreas is more likely to occur in young females with middle age, clinical presentations were as follows: a long-term persistent mass and distending pain in the epigastrium. Utrasounography, CT and ERCP could contribute to the diagnosis. Of the 11 cases, pancreaticoduodenectomy was performed in 2, distal pancreatectomy and splenectomy in 6, local excision of the mass in 2, and loop type cystojejunostomy in 1. The rete of resection was 90.9%. Of them, 1 case died of abdominal abscess and septicemia postoperatively. All the 10 cases who survived postoperatively were followed up. Among the 10 cases, 1 case of pancreatic cystoadenoma died of cardiac infarction 3 months after operation, and 1 case of cystoadenocarcinoma died of metastasis 51 months after tumor excision. The other 8 cases still alived well now. Conclusions Cystic tumor of the pancreas should be suspected when young or middle aged female patients have a long term big mass in epigastrium or left upper quadrant. Ultrasonography and CT scan are helpful in the diagnosis of the tumor including cystoademoma and cystoadenocarcinoma, Although the volume of the tumors may be very big, they often have a intact capsure,so easy to excise. The excision of the total tumor and some arounding pancreatic tissue is the first choice of the treatment for cystic tumor of the pancreas, and the curative effectiveness is satisfctory.
ABSTRACT
Objective: To select anti-HAb18G (hepatoma associated antigen) human Fd fragments with guided selection of L chain of chimeric Fab-HAb18. Methods: The human Fd repertories genes were amplified by RT-PCR from PBMC of hepatoma patients, and cloned into the vector pComb3X with chimeric L chain to construct the human-mouse hybrid Fab phage library. HAblSGE, extracellular region of HAblSG, was used as antigen to screen. The positive clones was obtained by ELISA and FCM with p Ⅲ-fusion Fab antibody. The DNA sequences were analyzed. Results: A human-mouse Fab antibody library were constructed with 2?107 PFU. After 6 rounds panning, 7 positive clones were obtained with ELISA and FCM. And sequences of 2 clones with better affinity were same. The CHI belonged to the IgG2 class as the parent Fd, and the variable region belonged to VH3 family. Conclusions: Through the construction of the HuMFab phage antibody library and chemeric L chain-guided selection, we get the available human Fd fragments for subsequent research.
ABSTRACT
0.05). RFA showed a marked short term therapeutic effect for patients with PHC.Conclusions After RFA treatment of patients with PHC,the serum levels of TNF and AFP decreased,and this suggests an augmentation of organism immune function. The serum levels of TNF and AFP can assist in the assessment of the therapeutic effect and prognosis of primary hepatic carcinoma.
ABSTRACT
Objective To introduce how to improve the operation quality and clinical effect of pancreatoduodenectomy (PD). Methods The morbidity and mortality of 126 patients with pancreatic head cancer or peri ampullary cancer treated by PD of traditional Child method and modified Child method ( pancreatic stump closing style pancrea to jejunum anastomosis) in our department from 1973 to 2002 were analyzed retrospectively. Results In modified Child method group, the morbidity of pancreatic leakage and the mortality of the operation were 2.4% and 4.7% respectively, which were significantly lower than those in the traditional method group(P