Objective To establish a method of isolating,culturing,proliferating urinary tract(nephropelvics,ureter and bladder)smooth muscle cells (SMC). Methods Urinary tract SMC were isolated from sample tissues (1 from human nephropelvics,1 from human ureter,3 from human bladders,5 from porcine bladders and 1 from porcine ureter) by collagenase digestion and were cultured in DMEM supplemented with fetal bovine serum.Morphology and expansion of the cells were observed.SMC specific protein (?-actin) was identified by immunohistochemical methods. Results The cells grew well and presented a typical morphological feature of SMC.They could be passaged 6 times without a remarkable decrease in rate of cell proliferation.Immunohistochemical staining showed that ?-actin was positive. Conclusions This urinary tract SMC cultural method was simple,reliable and fruitful.

Objective To study the biological effect of ?3 adrenoceptor-activated electrophysiological activities in cardiac myocytes. Methods Changes of transient outward potassium ( Ito) and L-type calcium current ( ICa-L) in primary cultured guinea pig cardiacmyocytes were detected with the standard whole-cell patch clamp technique about 5 -10 min before and after administration of ?3 adrenoceptor agonist,4--2-hydroxy-( 3-chlorophenyl) ethyl-aminopropylphenoxyacetate ( BRL-37344; BRL). Results BRL could significantly increased the Ito in guinea pig cardiac myocytes with the I-V curve up-moved. When the membrane potential reached to + 80 mV,the density current increased from 6. 11 ? 1. 03 pA/pF to 8. 46 ? 2. 07 pA/pF ( n = 6,P =0. 013 064). BRL ( 10 -6mol/L) could significantly increase the ICa-Lin guinea pig cardiac myocytes,which was ( 2. 30 ?0. 75) -fold higher than in control group( n =5,P =0. 0063). When the membrane potential increased to + 10 mV ,the current,increased from 89. 25 ? 17. 83 pA to 205. 00 ? 72. 24 pA ( n = 5,P = 0. 006 3). Conclusion BRL-37344 can increase the transient Itoand ICa-L,thus further regulating the cardiac activities.

Acetylcholinesterase ; analysis ; Antibodies, Monoclonal, Murine-Derived ; metabolism ; Australia ; Biopsy ; Calbindin 2 ; analysis ; Child ; Diagnosis, Differential ; Endoscopy, Gastrointestinal ; Eosinophilic Esophagitis ; pathology ; Hirschsprung Disease ; metabolism ; pathology ; Humans ; Intestinal Diseases ; pathology ; Lymphangiectasis, Intestinal ; immunology ; pathology ; Multiple Endocrine Neoplasia Type 2b ; pathology ; Nervous System Diseases ; pathology ; Quality Control

Objective To investigate vascular endothelial growth factor (VEGF),its receptor KDR and angiogenesis in relation to progression of colorectal carcinoma (CC). Methods KDR,VEGF protein expression and microvessel density (MVD) in 102 cases of human CC were examined by immunohistochemical staining. MVD,KDR and VEGF expression were analysed with their relation to histological grade,depth of invasion,Dukes stage,lymph node metastasis,liver metastasis and prognosis of CC. Results MVD was significantly higher in KDR and VEGF-positive CC than in KDR and VEGF-negative CC ( P

ObjectiveTo investigate the clinical and pathological features of Hirschprung disease (HD), intestinal neuro-nal dysplasia (IND) and hypoganglionosis (IH) in children.MethodsThe clinical data and pathologic slices from 238 children with intestinal dysganglionosis were retrospectively analyzed. The age, sex, involved intestinal length of children and prognosis were compared.ResultsIn 238 patients, 138 (58.0%) were diagnosed by rectal mucosal biopsies. There were 122 HD patients whose median age at diagnosis was 9 months and the ratio of male to female was 4.3:1, without involvement of whole colon. There were 45 IND patients whose median age at diagnosis was 14 months and the ratio of male to female was 1.05:1, and the whole colon of 33.3% patients was involved. There were two male IH patients whose ages at diagnosis were 12 years and 18 years respectively, and their whole colon was involved. There were 59 patients with HD complicated by IND whose median age at diagnosis was 13 months and the ratio of male to female was 5.56:1 and the whole colon of 16.9% patients was involved. There were 10 male patients with HD complicated by IH whose median age at diagnosis was 11.5 months and the whole colon of 80.0% patients was involved. The ages at diagnosis, the sex ratio, the rates of whole colon involved, and the cure rates among 5 groups were signiifcantly different (allP<0.01).ConclusionsThe rectal mucosal biopsy was the main method in diagnosis of intestinal dysganglionsis in children. Patients with HD had higher incidence and mild condition and favorable prognosis. Patients with IH or patients with HD complicated by IH had lower incidence rates and severe condition and poor prognosis, followed by patients with IND or patients with HD complicated by IND.

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

Objective To explore the clinicopathological characters of T2 gastric cancer and the value of MSCT in the preop‐erative TNM staging of T2 gastric cancer .Methods A total of 93 patients with T2 gastric cancer were included in our study and un‐derwent preoperative MSCT staging ,who were confirmed by pathologic results .Then the results were compared with those of path‐ologic TNM staging .Also the clinicopathological features of the T2 gastric cancer were analyzed .Results There were no statistical‐ly significant differences in the clinicopathological characters among T2a and T2b patients (P>0 .05) .Comparing with pathologic TNM stage ,the T staging accuracy of MSCT was 91 .40% (85/93) ,the N staging accuracies of CT was 66 .67% (62/93) ,in which , 68 .18% (30/44) ,65 .00% (26/40) ,60 .00% (3/5) and 75 .00% (3/4) were for pN0 ,pN1 ,pN2 and pN3 .And the TNM staging ac‐curacies of CT was 67 .74% (63/93) ,in which ,68 .18% (30/44) ,64 .10% (25/39) ,60 .00% (3/5) and 100% (5/5) were for stageⅠ ,Ⅱ ,Ⅲ and Ⅳ .Conclusion There are no significant different on clinicopathology features among T2a and T2b patients .MSCT can clearly determine the preoperative TNM staging of T2 gastric cancer .

BACKGROUNDTo evaluate the efficacy and side effects of weekly administration of gemcitabine combined with docetaxel in the treatment of advanced non-small cell lung cancer.

METHODSTwenty-six patients with advanced non-small cell lung cancer, with or without prior chemotherapy, were entered into this study. Gemcitabine and docetaxel were administrated weekly for 3 consecutive weeks, followed by 1 week rest. Gemcitabine was given as 800-1 200 mg/m² by intravenous infusion on days 1, 8, 15; while docetaxel was 35 mg/m² intravenously on the same days as gemcitabine. The efficacy including response rate and median survival duration and toxicity were observed.

RESULTSOf the 26 patients, one achieved complete response (CR), and 6 achieved partial response (PR), with an overall response rate of 27%. The median survival duration was 9.5 months and 1-year survival rate was 38% (10/26). The main toxicities were neutropenia and thrombocytopenia. One patient died from allergic shock.

CONCLUSIONSThe combination of docetaxel and gemcitabine is effective and well-tolerated in the treatment of advanced NSCLC.

OBJECTIVETo study clinicopathologic features, immunohistochemical profile, diagnosis and differential diagnosis of childhood central nervous system primitive neuroectodermal tumors (CNS PNETs) with the features of ependymoblastoma and neuroblastoma.

METHODSThe clinical data, morphologic and immunohistochemical features were analyzed in 4 cases of pediatric CNS PNETs with features of ependymoblastoma and neuroblastoma. EnVision method immunohistochemistry was applied.

RESULTSFour patients including three boys and one girl presented at the age from 12 month to 4 years and three tumors located in cerebrum, one in brain stem. All tumors showed typical combined histological patterns of ependymoblastoma and neuroblastoma, demonstrating zones of true rosettes, occasional pseudovascular rosettes, and undifferentiated neuroepithelial cells in a prominent background of mature neuropils. There was focal expression of glial fibrillary acidic protein (GFAP) consistent with glial differentiation and epithelial membrane antigen (EMA) consistent with ependymal differentiation. Necrosis was seen in three cases and calcification was present in one case. Immunohistochemically, the rosettes and undifferentiated neuroepithelial cells were positive for vimentin, partially positive for GFAP and EMA but negative for synaptophysin. The tumor cells were also positive for synaptophysin in neuropils. The Ki-67 label index ranged from 20% to 60%.

CONCLUSIONSCNS PNETs with the features of ependymoblastoma and neuroblastoma is a rare tumor with poor prognosis. The tumor primarily occurs in childhood, especially infant and belongs to the family of embryonal tumors of the CNS. The morphologic, immunohistochemical and genetic features are important in differential diagnosis from other tumors of the CNS.

Antigens, Neoplasm ; metabolism ; Central Nervous System ; pathology ; Child ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Infant ; Male ; Mucin-1 ; metabolism ; Neuroblastoma ; diagnosis ; pathology ; Neuroectodermal Tumors, Primitive ; diagnosis ; pathology ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; pathology ; Synaptophysin ; metabolism ; Vimentin ; metabolism