Objective To investigate the diagnostic value of contrast-enhanced ultrasound (CEUS) guiding normal saline (NS) injection through endoscopic nasobiliary drainage duct(ENBD) on evaluation for residual stones in common bile duct.Methods Fifty-five patients with bile duct stones were treated by endoscopic retrograde cholangio-pancreatography (ERCP) and duodenoscopic sphincterotomy incision surgery (EST) and ENBD.All patients received normal ultrasonography and CEUS guiding NS injection ultrasonography after EST.The length and width of common bile duct and the detection rate of residual stones before and after NS injection were compared.Results In the 55 patients,1 patient failed in injection of contrast agent into the ENBD.In the other 54 patients,the difference of the length and width of common blie duct before and after NS injection were statistically significant [(2.94±1.76)cm vs (6.09±1.46)cm,(0.58±0.30)cm vs (1.11±0.98)cm](all P<0.001).The full display rate of the common bile duct before and after NS injection were 13.0%(7/54) and 90.7%(49/54),respectively.Before injection,none of common bile duct stones was suspected.After injection,5 cases of common bile duct stones were suspected.Three cases were confirmed by ERCP,1 case was confirmed by operation and 1 case was false positive.Conclusions CEUS of the common bile duct through ENBD performs its patency and course.On this basis the injection of NS increases the display rates of common bile duct,thus improving the detection rate of residual stones.

Objective　To study the efficiency of the treatment of experimental hepatocellular carcinoma in rats with arsenic trioxide and to elucidate the possible mechanism.Methods　Wistar rats were fed with diethylnitrosamine (DEN) to induce HCC, then treated with As2O3. The histological changes in liver tissue were observed under microscope, and the cellular dynamic parameters were studied by flow cytometry. Results　Treatment with As2O3 caused HCC cells death via both apoptotic and non-apoptotic mechanisms when the dose was high (5 mg/kg), the necrosis was seldom and apoptosis was common when the dose was appropriate (1 mg/kg). Proliferation index (PI) decreased sharply in high-dose (5 mg/kg) group (P＜0.01), but not in other two (1 mg/kg, 0.2 mg/jg) groups (P＞0.05). However, S phase fraction (SPF) decreased dramatically in all three groups (P＜0.01). Although apoptosis of HCC cells was common in all three groups, it reached the top only when the dose (1 mg/kg) was appropriate (P＜0.001), and it was obviously accompanied with accumulation of cells in G2/M (G2/M restriction). Conclusion　These date demonstrate that arsenic trioxide induces apoptosis of rat HCC cells, and it is closely associated with G2/M restriction when apoptosis reaches the top. The data also suggest that arsenic trioxide can inhibit cell proliferation, which is dose-dependent and time-dependent. The fact that continuous intermittent i.p. injection of arsenic trioxide can also be effective may afford a novel way to use the drug more safely.

BACKGROUND:Mesenchymal stem cells have a extreme prospect in orthopedics, which show great potential especially in the treatment of articular cartilage defect disease. Bone marrow is the main source of mesenchymal stem cells, and the iliac puncture is a conventional way to obtain bone marrow, but is restricted by the limited resources and strict technical requirements. Therefore, it is of great significance to explore new effective and convenient sources of mesenchymal stem cells. OBJECTIVE:To explore the feasibility of autologous mesenchymal stem cells derived from the joint drainage fluid after knee arthroscopy.METHODS: We selected eight patients who underwent arthroscopic surgery to collect joint drainage fluid by pre-made sterile blood bag before the wound closure. Precipitation with hydroxyethyl starch and density gradient centrifugation method were performed to isolate and culture mesenchymal stem cells from the joint drainage fluid. Cell morphology, growth curve, surface marker identification were observed and detected using flow cytometry. Then, adipogenic, chondrogenic and osteogenic differentiation of cells were induced and identified by oil red O, toluidine blue staining, and alizarin red staining, respectively. RESULTS AND CONCLUSION:The cultured cells were spindle-shaped, adherently grew and had good proliferation ability, which were positive for CD44, CD90, CD105 and CD73, but not for CD45. Under standard inductions, the cultured cells were induced to differentiate into osteoblasts, adipocytes and chondrocytes. Therefore, these cells were confirmed as mesenchymal stem cells. Mesenchymal stem cells were successfully isolated from the joint drainage fluid of eight patients and had no difference in cell morphology, proliferation and phenotypes. To conclude, the joint drainage fluid is an ideal source of mesenchymal stem cells with the guaranteed quality and quantity.

Purpose To investigate bax expression and induction of apoptosis in normal cultured retinal pigment epithelium (RPE) cells . [WT5”HZ]Methods [WT5”BZ]Cultured human RPE cells were transfected by P MDNA3 hbax ,which incoded the whole bax gene and may be induced by Zn 2+ under the MTII promoter, through lepofectin mediated protocol.The tested RPE cells were divided into three groups of A, P MDNA3 hbax transfected ;B,P MDNA3 (nude vector) transfected and C,normal RPE cells.After transfection, DNA gel electrophoreses were performed ,the tested RPE cell cycles were analyzed with flow cytometry (FCM). Results The gel electrophoretogram showed DNA ladder phenomenon,FCM confirmed the apoptosis of RPE cells P MDNA3 bax transfected , consisting of significant apoptotic peak sited before the G 1 phase and the apoptotic rate was 36%. Conclusion The foreign bax gene can be effectively conducted into the RPE cell through lepofectin mediated protocol and induced expression . The foreign bax overexpression may induce the cultured human RPE cell susceptibility to apoptosis.

Objective To establish a reconstructed human epidermis model in vitro, and to study the process of re-epithelialization. Methods A dermal substrate devoid of epidermis was prepared; a 2 mm skin biopsy explant was transplanted onto the dermal substrates. Visualization of epidermal cell migration was carried out by fluorescence imaging. The proliferation and differentiation of the new epithelial cells were observed using histopathological and immunohistochemical staining (Ki67). EGF was added to the culture medium of the experimental samples but not to that of the controls. Results After 3 days of culture, re-epithelialization was observed on the surface of the dermal substrate. A complete structure resembling regular epidermis was noted in 10 days. As compared to control samples, EGF-treated samples had larger area of re-epithelialization (t= 3.02, P

BACKGROUND:Vascular endothelial growth factor and transforming growth factor play a crucial role in embryonic development, wound healing, inflammation, cancer, ischemic hypoxia and other physiological and pathological processes, and participate in the development and progression of brain damage. OBJECTIVE: To evaluate the differences in the expression of vascular endothelial growth factor and transforming growth factor-α during the early phase of cerebral aneurysm formation in rats. METHODS:Twenty-eight healthy Sprague-Dawley rats were randomized into three groups. Sham operation group (n=8): the left carotid artery bifurcation and bilateral renal artery were only exposed, without ligation, and rats were kiled that day. 15 days group (n=10) and 30 days group (n=10): the left common carotid artery, internal carotid artery, external carotid artery and bilateral renal artery were ligated, to establish aneurysm model, and rats were kiled at 15 and 30 days, respectively. The bilateral sides of the anterior cerebral artery/olfactory artery bifurcations were harvested and observed under light microscopy for pathological changes. Immunohistological staining was performed to detect the expression of vascular endothelial growth factor and transforming growth factor-α. RESULTS AND CONCLUSION:The results showed that, no aneurysm formed in the sham operation group and 15 days group. In the 30 days group, one saccular aneurysm and five early aneurysm-like changes were found in the right anterior cerebral artery/olfactory artery bifurcations. In the sham operation group and 15 days group, no vascular endothelial growth factor was expressed. In the 30 days group, the positive rate of vascular endothelial growth factor was up to 80%, indicating that vascular endothelial growth factor is possibly involved in the formation of aneurysm. Transforming growth factor-α expression in the sham operation group and 15 days group was more apparent than that in the 30 days group, indicating that transforming growth factor-α is damaged or secretion is reduced in this process, which was possibly related to the formation of aneurysm.

Objective To investigate the efficacy and safety of different acoustic power of high-intensity focused ultrasound (HIFU) in treating human pancreatic xenograft models. Methods Human pancreatic cancer cells (YY-1) were implanted subcutaneously in nude mice to establish animal models. The tumor bearing mice were divided into low-power HIFU treatment group (200 W,n=10), high-power HIFU treatment group (300 W,n=10) and blank control group (n=10). The change of tumor volume, the tumor growth rate and side effects were recorded. The apoptosis rate of tumor cells of each group was determined by TUNEL method. Results The tumor volume and the tumor growth rate of the low-power group and the high-power group were significantly lower than those of the control group (P<0.05), while no statistically significant differences in the tumor volume and the tumor growth rate existed between the low-power group and the high-power group (P>0.05). Compared with the low-power group, the incidence of side effects in the high-power group was significantly higher (P<0.05), including mainly skin burn (60%) and acoustic channel injury (20%). At the 7th and 14th day after the treatment, the apoptosis rates of tumor cells in both the low-power group and the high-power group were significantly higher than that of the control group (P<0.05), but the difference in the apoptosis rates of tumor cells was not statistically significant between the low-power group and the high-power group (P>0.05). Conclusion For the treatment of human pancreatic xenograft tumor in nude mice models, HIFU with low power is effective and safer.

BACKGROUND:Study confirmed that the de-epidermized dermis(DED)can be used as dermal substitute and may form epidermal structure after incubating keratinocytes.However,the cell biological activity,tissue structure characteristics and the basement membrane component analysis of dermal substitute have been reported less.OBJECTIVE:To investigate the cell activity and the tissue structure characteristics of DED.METHODS:Skin flap was treated with 56℃ phosphate buffered solution to remove the epidermis,and the dermal cell components were deleted by freezing and thawing with liquid nitrogen to obtain DED.The DED cell activity was detected with tissue culture method,hematoxylin nuclear staining was used to determine the DED cell nuclei,and vimentin immunohistochemistry was applied for fibroblast determinations.The basement membrane and its components were detected using Periodic Acid-Schiff staining and collagen type Ⅳ immunohistochemistry.Van Gieson stain,Weigart stain and those double staining were respectively used to determine DED collagen fibers and elastic fibers.The DED ultrastructure was observed under transmission and scanning electron microscope.RESULTS AND CONCLUSlON:Using tissue culture method,the cultured DED did not exhibit cell growth at 2 weeks.Hematoxylin-eosin staining showed no nuclear in DED,vimentin immunohistochemistry showed no vimentin expressed in DED.Van Gieson staining showed DED collagen fibers were stained as rose red,Weigert staining showed DED elastic fibers were stained as pu rplish black double staining further demonstrated uniform arrangement of collagen fibers and elastic fibers.DED surface and the remaining appendages were strongly positive for Periodic Acid-Schiff staining,and type Ⅳ collagen expression was significant.Transmission and scanning electron microscope results showed that,the DED elastic fibers and collagen overlap arranged with pore intervals,they intercrossed into a network.There is no living cell component in DED,dermal matrix surface and appending organ luminal wall still retain glycogen,type Ⅳ collagen and other basement membrane components,dermal matrix is rich in collagen and elastic fibers.it is a three-dimensional collagen matrix similar to in vivo dermis.

Objectives To explore the clinical diagnosis and treatment of Dieulafoy disease in children. Method The clinical features, endoscopic features and treatment of Dieulafoy disease in a child were reviewed. Results The 2-year-5-month old girl was admitted due to hematemesis for 7 hours. She was diagnosed of Dieulafoy disease by the typical endoscopic appearance. Gastroscopy showed that the lesion was located in gastric angle which was the predilection position of Dieulafoy disease. The small red blood vessels in the central part of the erosion area was exposed on the mucosal surface. The high frequency electrocoagulation under gastroscope was performed and effect was definite. Conclusion Dieulafoy disease is rare in children and lacks obvious clinical features. Endoscopic treatment has definite effect with little trauma and is the first choice of treatment.

BACKGROUND:Matrix protein is an essential component of the vascular wal , provides a necessary frame for the integrity of the vessel wal and physiological function of vascular wal cel s, and regulates cel s and smooth muscle. OBJECTIVE:To construct rat model of early aneurysm, and to evaluate differences in the expression of matrix structural proteins during cerebral aneurysm formation. METHODS:Twenty-eight healthy male Sprague-Dawley rats were randomized into control group (n=8) and model group (n=20). Aneurysm model was established by ligation of the left common carotid artery and right renal artery-induced hypertension in the model group. In the control group, only the left carotid artery bifurcation and bilateral carotid were exposed in rats. Rats in the model group were sacrificed at 15 and 30 days after model establishment. Right anterior cerebral artery in rats and olfactory artery bifurcation received immunohistochemical staining. The expressions of fibronectin,α-smooth muscle actin and col agen III were analyzed. RESULTS AND CONCLUSION:Compared with the control group, no significant difference in fibronectin expression was detected in right anterior cerebral artery and olfactory artery bifurcation in rats of the model group at 30 days after model establishment (P>0.05). However,α-smooth muscle actin and col agen III expressions were significantly reduced (P<0.05). These data confirmed that expression of structural proteins had differences and dynamic changes during early aneurysm formation in rats. Degradation of matrix structural protein in cerebral artery may be one of the key mechanism of aneurysm formation.