Objective To summarize the clinical and pathological characteristics of gastrointestinal stromal tumors with hepatic metastasis, and to analyze its survival and explore its principles of diagnosis and treatment. Methods Among 99 patients diagnosed as gastrointestinal stromal tumors who had a completely case history in our hospital, we retrospectively analyzed the clinical data of 26 patients with hepatic metastatic and the factors influencing survival. Results The average age at diagnosis of primary and hepatic metastatic gastrointestinal stromal tumors was 50.8 and 51.8 years old respectively. Five cases were confirmed by pathological examination, 12 cases were diagnosed by the exploration during the operation and 14 patients had an imaging diagnosis only. Synchronous and metachronous hepatic metastasis happened in 8 and 18 patients respectively. The median interval between the primary tumor and the metachronous hepatic metastasis was 12 months. The primary sites of 12 cases were in stomach, 5 in colorectum, 6 in small intestine and 3 in extra-gastrointestinal tract.Four cases of the hepatic metastatic tumors were treated with surgical resections, 2 with injections of anhydrous alcohol, 3 with interven-tional therapies, 7 with systemic chemotherapies, 8 with imatinib and 2 without treatment. The median survival was 21 months after hepatic metastasis. The administration of imatinib was an important factor prolonging the survival after hepatic metastasis. Conclusions The most frequent primary site of hepatic metastatic stromal tumor is the stomach while small intestinal stromal tumors are most inclined to metastasize to the liver. Treatment with imatinib for more than 3 months can prolong the survival.

Objective To study the proliferation and migration of vascular smooth muscle cell (VSMC) after transferred angiotensin Ⅱ (AngⅡ) type 2 receptor (AT2R) gene. 　Methods The recombinant adenoviral vector, AdCMV-AT2R, containing rat AT2 receptor gene was constructed by homologous recombination, and transfered to rat VSMC in vitro. The expression of AT2R mR NA was detected by RT-PCR. The rate of expression and the change of cell cycle in VSMC were analysed by flow cytometry. These assays including cell devision index. Incorporation of bromodeoxyuridine(BrdU) and 3-(4,5-dimethyl-thiazol-2-yl)2,5-diphenyltetrazolium bromide( MTT) were used to determine the proliferation of VSMC. The modified Boyden's chamber method was used to test the migration of VSMC. The r e-organization of F-actin in VSMC was analysed by confocal microscopy. 　Results RT-PCR showed that the expression of AT2R mRNA increased substantially in transferred VSMC, and the peak value of expression rate is about 89.51% at 48 hours. When the expression of AT2R is at peak value, the ratio of S, G2 and M periods was reduced from 31.7% to 13.9%(P<0.05). The OD values of MTT and BrdU incorporation were reduced by 61.4% and 51.6% respectively(P<0.0(1). the number of VSMC migration was reduced by 62.2%(P<0.05) and the expression of F-actin was also decreased signific antly. 　Conclusion AdCMV-AT2R induced high level expression of AT2 receptor in cultured rat VSMC and its expression can significantly inhibit the proliferation and migration of rats VSMC in vitro.

BACKGROUND:The benefit of cell therapy may be partly due to the secretion of angiogenic and antiapoptotic growth factors.Whether amniotic fluid stem cells (AFS) could secrete some growth factors requires further studies.OBJECTIVE:To isolate and culture AFS cells,and explore the angiogenic or antiapoptotic effect of cytokines secreted by AFS on endothelial cells.DESIGN,TIME AND SETTING:A in vitro cytological experiment was performed at the Institute of Hypertensive Disease,First Affiliated Hospital,Nanchang University from December 2008 to June 2009.MATERIALS:Term amniotic fluid of ten samples,50 mL/case,was obtained following caesarean delivery.The umbilical vein was used to isolate endothelial cells.Written informed content was obtained from all women.METHODS:AFS isolated from human amniotic fluid was cultured and digested by trypsin at confluence of 80%.The third passage of cells at a density of 5×10~8/L were divided into two groups:hypoxia group:the cells were cultured in 2% O_2 + 5% CO_2 +93% N_2;normal group:the cells were cultured in 5% CO_2 + 95% air.Two groups were cultured at 37 ℃ for 24 hours.The supematant of two groups was collected.The second passage of human umbilical vein endothelial cells cultured in vitro was collected and seeded onto 12-well culture plate at a density of 2×10~4 cells/well,and divided into 3 groups:control group was cultured in 2 mL EBM-2 containing 5% fetal bovine serum (FBS);normal group was cultured in 1 mL EBM-2 containing 5% FBS and 1 mL AFS cell culture solution;hypoxia group was cultured in 1 mL EBM-2 containing 5% FBS and 1 mL hypoxia AFS cell culture solution for 3 days,followed by incubation with 10 μg/L tumor necrosis factor (TNF)-α.MAIN OUTCOME MEASURES:AFS surface phenotype was examined by flow cytometry;the secretion level and mRNA expression of vascular endothelial cell growth factor (VEGF) and hepatocyte growth factor (HGF) were examined by ELISA or RT-PCR.The proliferation and apoptotic rates of endothelial cells were examined.RESULTS:AFS cells were long fusiform-shaped and arranged radially after 7 days of culture.The third passage of AFS cells expressed CD29 and CD105 while did not express CD34.AFS cells of normal culture secreted VEGF and HGF;AFS cells of hypoxia condition significantly increased secrete of VEGF (P＜0.01),and VEGF mRNA expression was significantly upregulated (P＜0.05),while HGF and mRNA expression remained unchanged (P＞0.05).Compared with control group,the number of endothelial cells was significantly increased in normal and hypoxia AFS cell groups after 3 days of culture (P＜0.05).After cocultured with TNF-α for 24 hours,the apoptosis rates of endothelial cells in AFS-conditioned medium was significantly decreased (P ＜ 0.05),and the change degree of hypoxia AFS cell group was greater than normal AFS cell group (P ＜ 0.05).CONCLUSION:AFS can secrete cytokines such as VEGF and HGF.Moreover,it significantly promotes endothelial cells proliferation and inhibits apoptosis.Under hypoxia condition,the secretion of VEGF from AFS cells is increased,and the effects on endothelial cells proliferation and apeptosis are enhanced.

AIM:A new method for the overall quality control of Radix Scutellariae has been established by HPLC digitized fingerprint. METHODS: The chromatographic fingerprints were obtained from injecting 5 ?L of the sample solution each time on a CenturySIL C_(18) BDS column(200 mm?4.6 mm,5 ?m) with the gradient elution solvent system composed of 1% acetic acid water and 2% acetic acid acetonitrile.The flow rate was 1 mL/min,the column temperature was maintained at(30?0.15)(?C) and the detection wavelength was set at 280 nm.The chromatographic fingerprints were evaluated by the software of the digitized evaluation system of Traditional Chinese Medicine fingerprint with the super information characteristics. RESULTS: 30 co-possessing peaks were selected as the fingerprint peaks of Radix Scutellariae and the similarities between the chromatographic fingerprints and the herbal drugs were calculated by taking baicalin peak as the referential peak.The chromatographic fingerprints were also evaluated by the chromatographic fingerprint index(F). CONCLUSION: This method with good precision and reproducibility can be applied to the quality control of Radix Scutellariae.

AIM:To establish the assessing method of traditional Chinese medicine chromatogrphic fingerprints by the involution similarity and quantitative involution similarity and its application. METHODS: The fingerprint of Ginkgo biloba L.extract(GBE) was evaluated by the involution similarity and quantitative involution similarity,and the contrast studies were implemented by other appraisal targets,eg.the included angle cosine S_F,the correlation coefficient r and the indexes with quantitative characteristic,such as the apparently quantitative similarity R%、the average mass percent M%,etc.. RESULTS: The involution similarity could reflect the similarity change in each sample from different bathes and be of quantitative function.The quantitative involution similarity and quantitative weighted similarity could perfectly reflect the changes in contents of the consituents of GBE. CONCLUSION: The involution similarity and quantitative involution similarity can qualitatively and quantitatively assess the similarity between sample and the referential fingerprint.They are certainly the novel method of evaluating the traditional Chinese medicine chromatographic fingerprints.

AIM: To observe the dynamic changes of interleukin-4(IL-4),IL-10,IL-12 in rat serum and lung tissues during acute respiratory distress syndrome (ARDS). METHODS: The ARDS model of rats was induced by intravenous injection of oleic acid. The levels of IL-4 ,IL-10,IL-12 in serum and the supernatant of lung tissues were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: The Levels of serum and lung IL-10,IL-12 in ARDS rats were increased in 4 h ,8 h,16 h group compared with control group . The levels in IL-10 in serum in 16 h group and IL-10 in lung tissues of 8 h group were lower than that in 4 h group. The Levels of IL-4 in serum in 4 h, 8 h group were higher than that in control group , while IL-4 in 16 h group was lower than that in 8 h group. IL-4 of lung tissues in 4 h,8 h,16 h group were increased significantly,but in 16 h group were lower than that in 8 h group. The biggest changes of pulmonary coefficient and histopathology were observed at 4 h after injection of oleic acid. CONCLUSIONS: IL-4,IL-10 and IL-12 might play important roles in inflammatory reaction induced by oleic acid. The pro- and anti-inflammatory cytokines produced successively during ARDS. The relationship between unbalanced cytokines and lung injury in ARDS needs to be further studied.

Objective To summarize the successful experience of three cases of heart-lung transplantations performed in our institute.Methods From July 2003 to August 2012,three patients,with diagnosis of end-stage heart-lung diseases,received heart-lung transplantation in our institute.One case was diagnosed as congenital atrial septal defect,Eisenmanger syndrome,NYHA class Ⅳ; one was dilated cardiomyopathy with moderate/severe pulmonary arterial hypertension,NYHA class Ⅲ-Ⅳ,one was diagnosed as double outlet left ventricle (DOLV) with ventricle septal defect and stenosis of pulmonary artery and its left and right branches,NYHA class Ⅲ-Ⅳ.Donor hearts were preserved with UW solution,donor lungs were preserved with Euro-Collin solution in case one and with low potassium dextran containing prostaglandin E1 in the others.Extensive disinfection and strict scrutiny were implemented postoperatively.Immunosuppressive therapy included administration of zenapax or basiliximab preoperatively,methylprednisolone during the operation,and cyclosporine a/tacrolimus + prednisone + mycophenolate postoperatively.Surgical hemostasis is of great importance,as the total pleural effusion reaches 14 640 ml within 31 days postoperatively in case two.Strict postoperative disinfection and isolation were implemented,and management of the respiratory tract was intensified.Therapeutic bronchoscopy was performed frequently for sputum suction.In case two,bronchoscopy was used thirteen times within 40 days after transplantation.Broad-spectrum antibiotics and antifungal antibiotics were used for infection control.Results All three patients were discharged after recovery from operation.Case one died of obstructive bronchitis and lung failure caused by chronic rejection four years and ten months postoperatively.Case two died of sudden cerebrovascular accident 68 days after operation.Case three survives more than one year postoperatively so far and is still alive.Conclusion Proper preservation of the donor heart and lung,perfect surgical hemostasis,strict infection control,frequent application of bronchoscopy and appropriate immunosuppressive management are critical to the success of heart-lung transplantation.

Objective To constructed the cell model transferred angiotensin Ⅱ (AngⅡ ) type 2 receptor (AT2R) in vascular smooth muscle cells(VSMC) Method The VSMCs, isolated from the aorta of rat, were cultured by routine method. Recombinant adenoviral vector, AdCMV-AT2R, containing rat AT2 receptor gene was constructed by homologous recombination, and then it was used to transfer AT2 receptor gene to VSMC in vitro. The rate of AT2R expression in VSMC was analysed by flow cytometry. The expression of mRNA, protein were detected by RT-PCR, Western blot and immunohistochemistry respectively. The angiotensin Ⅱ (AngⅡ) type 1 receptor (A T1R) was determined as well. Result The expression rate of AT2R in VSMC was increased signific antly after transferred by AdCMV-AT2R with time, and the peak value detected by flow cytometry was about 89.51% at 48 hours. RT-PCR, Western blot and immunohistochemistry showed that the expression of AT2R mRNA and protein were increased obviously in transferred VSMC. There were no significantly change of AT1R expression during AT2R expression. Conclusion Our study indicates that AdCMV-AT2R did generate high level expression of AT2 receptor and its expression did not affect AT1R exp ression in cultured VSMC. The VSMCs transferred AT2R gene may be used as a good model to study the effect of AT2R on their biological action such as proliferation, migration and apoptosis.

Objective To evaluate the immediate and short term therapeutic efficacy of percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM) and to investigate the prevention of the related complications. Methods PTSMA was conducted in 3 patients with HOCM refractory to medication. The immediate effect was evaluated by pressure monitor, but the follow up effect by observation of the clinical symptoms and ultrasonic cardiogram. Results The left ventricular outflow tract gradients triggered by ventricular premature beats decreased from 143 mmHg (ranging from 70 to 180 mmHg) to 53 mmHg (ranging from 30 to 80 mmHg). Complete atrioventricular block (Ⅲ?AVB) with atrioventricular node rhythm was found in 1 case, and complete right branch bundle block (CRBBB) in another one. Follow up of the 3 cases at 6 months after operation revealed remarkable improvement of the patients' cardiac function (NYHA), disappearance of angina pectoris, and obvious attenuation of ventricular septal hypertrophy and SAM (systolic anterior motion of mitral) phenomenon. Conclusion PTSMA is an effective method with satisfactory short term effect for the patients with HOCM refractory to medication.

OBJECTIVE[corrected] To assess the specificity and applicability of direct PCR sequencing in the detection of point mutations in hepatitis B virus (HBV) associated with drug resistance.

METHODSSerum samples were obtained from 120 patients with hepatitis B treated with nucleoside analogus for at least 2 years to detect the point mutations in HBV genome in association with drug resistance using nested PCR and direct DNA sequencing.

RESULTSForty out of the 120 patients were found to have one or two point mutations associated with drug resistance, including 17 with L180M and M204V/I mutations (42.5%), 10 with M204V/I mutation (25%), 8 with N236T mutation (20%), 3 with L180M mutation (7.5%), and 1 with both A181V/T and N236T mutations (2.5%), and 1 with A181V/T mutation(2.5%).

CONCLUSIONDNA sequencing is a good method to detect all known point mutations associated with HBV drug resistance.

Adolescent ; Adult ; Aged ; Child ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Male ; Middle Aged ; Nucleosides ; therapeutic use ; Point Mutation ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Young Adult