0.05). Phenylephrine induced contractile responses in a concentration-dependent manner in the dorsal detrusor strips, but not in the ventral detrusor strips. S-doxazosin, R-doxazosin and rac-doxazosin at 1 ?mol?L-1 antagonized the phenylephrine-induced contractile responses competitively in the dorsal detrusor strips, and their pKB values were 7.44?0.19, 7.39?0.14 and 7.38?0.30, respectively. Three pKB values of doxazosin and its enantiomers were not significantly different from each other. Electric field stimulation produced a steady contractile response that was completely inhibited by tetrodotoxin at 0.3 ?mol?L-1. S-doxazosin, R-doxazosin and rac-doxazosin significantly inhibited the contractile responses to electric field stimulation in the dorsal detrusor strips of the rabbit urinary bladder(P0.05). However, S-doxazosin, R-doxazosin and rac-doxazosin did not affect the responses to electric field stimulation in the ventral detrusor strips. Conclusion The pKB value of S-doxazosin against phenylephrine-induced contraction via-adrenoceptors is same to R-doxazosin and rac-doxazosin, and the three agents are able to inhibit the adrenergic contraction induced by electric field stimulation in dorsal detrusor strips of the rabbit urinary bladder.

Objective To determine the effect of Vibrio vulnificus cytolysin (VVC) inducing ap-optosis in human umbilical vein endothelial cell(HUVEC) and its possible mechanism. Methods The en-tire vvhA gene that encoding VVC from V. vulnificus strain GTC333 was amplified by PCR and sequenced af-ter T-A cloning. E. coli BL21DE3pET-42a-vvhA, a prokaryotic expression system of the vvhA gene, was then con-structed. Ni-NTA affinity chromatography was applied to purify the target recombinant protein rVVC, and SDS-PAGE plus Bio-Rad Agarose Image Analyzor were used to measure the output of rVVC and to determine the purity of rVVC extract. The activity of rVVC dissolving rabbit erythrocytes was detected by hemolysis test. DPNH chromotometry and TphBNa chromotometry were performed to examine the contents of LDH and K+ in the supernatants of rVVC-treated HUVEC cultures, respectively. The effect of rVVC inducing apepto-sis of HUVEC was detected by flow cytometry, rVVC was labeled with FITC and the location of FITC-labe-ling rVVC in HUVEC was observed by laser canfocal microscopy. Results The cloned whA gene had 96.09% and 98.26% similarities of nucleotide and amino acid sequences compared to the corresponding se-quences in GenBank. rVVC, with a dosage of 1 μg/ml, could dissolve rabbit erythrocytes (P<0.01). 10 μg/ml rVVC was able to promote the increases of K+ content (P<0.01) but no change of LDH content could be found in the cell supernatants. HUVEC was apoptotic after the cell was treated with 1～100 μg/ml of rVVC for 2 h. In the 5～240 min duration of co-incubation of FITC-labeling rVVC and HUVEC, the rV-VC gradually moved from surface to inner side of the membrane and then entered the cytoplasms. When FITC-labeling rVVC treated HUVEC for 30 min, most of the rVVC was found to be intracellular location. Conclusion rVVC has cytolytic activity. VVC has an ability to enter HUVEC and causes injury of HUVEC via inducing apoptosis, which may be the major pathogenic mechanism of VVC.

Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.

Castleman Disease ; genetics ; pathology ; surgery ; Diagnosis, Differential ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Hyperplasia ; Lung ; pathology ; surgery ; Lung Diseases ; genetics ; pathology ; surgery ; Lymph Nodes ; pathology ; Middle Aged

Objectives To investigate the pathophysiologic mechanisms and the clinical character- istics and variations of nutcracker esophagus(NE).Methods Clinical data obtained from 22 patients with NE were retrospectively analyzed.Seven followed-up patients had esophageal motility,multi-channel electrogastrography(MEGG),the autonomic nervous system(ANS) and psychology tests and were com- pared to 10 healthy subjects(HS).Results①In NE group,12 patients had reflux symptom(55%),7 patients had chest pain(32%)and 3 patients had dysphagia(13%).There was no statistical difference in mean contraction amplitude(MCA) between patients with reflux symptom and chest pain.②Eight out of the 13 patients with NE who received 24 h pH monitoring were positive reflux,and 4 out of 17 patients had reflux esophagitis in endoscopic examinations.The symptoms were improved in 58%patients(7/12) by regular acid-suppression therapies.③There was no statistical difference beween NE and HS groups in dominant frequency and power of MEGG.However,the percentages of normal rhythm in preprandial and slow wave coupling in pre and postprandial of NE patients were significantly decreased than those in HS.④The ANS function in NE group had no statistical difference compared to those in HS.⑤In follow- up group,no difference was found before and after nitroglycerin sublingually.Four patients had depres- sion.Conclusions The clinical presentations of NE are vary.The symptoms of NE were poorly correla- ted with MCA,but partially correlated with GER,which may represent a special subtype of GERD. Gastric dysmotility and psychological factors might contribute to the pathogenesis of NE.

Objective To investigate the efficacy and safety of autologous cryopreserved platelet transfusion in the management of thrombocytopenia after chemotherapy in hematological malignancy.Methods A total of 40 patients diagnosed as hematological malignancy with complete remission were equally assigned into study group and control group.During chemotherapy interval in the study group,when platelet counts exceeded 120 × 109/L,autologous platelets were collected with CS3000 Cell Separator and cryopreserved at-80℃ with 5％ dimethylsulfoxide.When platelet counts dropped below 15 × 109/L after chemotherapy,autologous platelets were thawed with 40℃ water bath and transfused back to each patient.In the control group,when platelet counts dropped below 15 × 109/L after chemotherapy,allogeneic fresh platelets were transfused.Median loss during the freeze-thaw-wash procedure in study group was observed,and the 1 h,24 h corrected count increments(CCI)were calculated in the both groups.The hemostatic effects and adverse reactions were also observed.Results In the control group,1hCCI and 24hCCI were (19.3 ±6.1)× 109/L and(12.2 ± 7.0)× 109/L,respectively,with the effective rate of 80％ and the transfusion reaction rate of 45％.Totally 20 collection and transfusions were finished in the study group.A total of(3.4-8.5)× 1011 platelet were obtained in each collection.Platelet recovery after freezing and thawing was(73.51 ±9.03)％(62％-83％).1hCCI was(17.4±7.6)× 109/L,24h CCI was(10.5 ±5.8)× 109/L and the effective rate was 85％.There was no significant different between the two groups (P ＞ 0.05).The transfusion reaction rate was 15 ％,which was significantly lower than that of the control group(P ＜ 0.05).Meanwhile,adverse reactions were occurred less in the study group.Conclusion This study demonstrates that autologous cryopreserved platelet transfusions can be safely administered for supporting thrombocytopenia in hematological malignancy patients undergoing chemotherapy.

ObjectiveTo investigate the prognostic value of fluid responsiveness in patients with acute respiratory stress syndrome (ARDS).Methods Fifty-nine mechanically ventilated patients with ARDS were enrolled.Their stroke volume variations (SVV) were detected by pulse contour continuous cardiac output (PiCCO) analysis device,and then the patients were subsequently divided into fluid responsive group (SVV≥15％) and fluid non-responsive group (SVV ＜ 15％ ).The differences in 28-day survival,length of ICU stay and duration of mechanical ventilation between the 2 groups were compared.Thereafter,the 28-day survival of patients was analyzed by Kaplain-Meier survival model and the relationship between SVV and mortality within 28 days was analyzed by logistic regression model.Results In comparison with fluid non-responsive group,the 28-day survival of fluid responsive group was significantly increased (85.3％ vs.56.0％,P =0.012),the length of ICU stay was significantly shortened [( 13.1 ±5.2 ) d vs. (21.6±9.0) d,P=0.008],and the duration of mechanical ventilation was significantly shortened as well [( 11.4 ± 5.3 ) d vs.( 18.3 ± 4.9) d,P =0.022] ; Logistic analysis revealed that SVV ＜ 15％ increased the risk of 28-day mortality ( OR =4.82 ; 95％ CI:2.67 ～ 11.71,P =0.009 ).ConclusionsSVV-based fluid responsiveness can be served as a prognostic predictor for ventilated patients with ARDS.

Objective To explore the changes of pubescent immune response in the schizophrenia offspring induced by poly(I:C) during pregnancy and the effects on white matter.Methods The obtained pregnant rats were randomly divided into model group(n=6) and control group (n=5), receiving either poly (I:C) at a dose of 10 mg/kg diluted in 0.9％ NaC1 solution or vehicle solution alone (sterile pyrogen-free 0.9％ NaC1) on gestation day 9 (GD9).Immunohistochemical technique(IHC) was applied to detect the changes of microglias and astrocytes in the prefrontal cortex(PFC) and hippocampus(HC) of partly offsprings in the two groups at the sixth week,as well as Luxol fast blue(LFB) for the changes of white matter.The other offsprings of each group were selected for behavioral assessment at the eighth week.Results The results of prepulse inhibition test showed that PP2, PP4 and PP8 of model groups were significantly lower than that of the control group at young adult(P＜0.01).In passive avoidance test, and the T1 results of model group were significantly higher than those of the control group, the T results of model group were lower than those of control group (P＜ 0.01).Immunohistochemical results indicated that the number of microglias in the model group((264±33)/mm2, (271 ±38)/mm2) was significantly increased in PFC and HC than that in the control group((140±29)/mm2, (169±37)/mm2, P＜0.05) ,which was accompanied with significant morphological changes, while the OD value of astrocyte protein expression in the frontal lobe and hippocampus had no obvious difference between the model group and control group(P＞0.05).The OD value of LFB staining for myelin in the model group(0.29±0.02) was significantly decreased compared with that in the control group(0.33±0.03)(P＜0.01).Conclusion The young adult offsprings with prenatal infection present obvious schizophrenia-like behavior, meanwhile, the microglias activation and demyelination changes in white matter are observed,which provides more evidence for the relationship between immune response and white matter in the pathogenesis of schizophrenia.

AIM:To explore whether YAP protein is important in induced pluripotent stem cell ( iPSC)-induced cardiovascular progenitor cell and/or vascular smooth muscle differentiation .METHODS:Using episomal vector based reprogramming , we generated human iPSCs from donor fibroblasts .We used both this iPSCs and human H 1 embryonic stem cells to differentiate into vascular smooth muscle cells (VSMCs) through cardiovascular progenitor cells (CVPC).Western blotting, qPCR and immunofluorescence microscopy were used to check the expression of YAP and related genes during this differentiation process .RESULTS:The results showed that iPSCs expressed pluripotent stem cell markers, such as Oct4, Nanog, Sox2, TRA-1-60 and SSEA3, and could form teratoma in SCID mice.YAP was highly expressed in pluripotent stem cells , but dramatically decreased when CVPC differentiation started .YAP gradually increased dur-ing CVPC three-day differentiation.The TAZ and YAP binding partner TEAD1, but not TEAD2 and TEAD4, have similar expression pattern in CVPC differentiation .Immunofluorescence result confirmed that YAP was activated and accumulated in nucleus .Interesting-ly, both YAP and phosphorylated YAP expression decreased to very low level after CVPC differentiated into VSMCs in 7 days.TEAD4 and TAZ also decreased, while TEAD1, TEAD2 and TEAD3 expression did not change during VSMC differentiation .CONCLU-SION:YAP and TEAD1 expression increased during CVPC differentiation , while YAP and TEAD4 expression decreased from CVPC to VSMCs differentiation , which suggested YAP might have different function during diverse cell differentiation .

Objective To determine the clinical significance of pulmonary function test(PFT)in evaluating the features and severity of lung impairments associated with connective tissue diseases(CTD)by comparing the differences of pulmonary function test parameters among groups of CTD associated pulmonary disorders.Methods Cases of CTD associated pulmonary disorders were prospectively enrolled and assigned into 3 groups according to their lung impairments:CTD associated pulmonary arterial hypertension group (CTD-PAH,n=29),CTD associated interstitial lung disease group(CTD-ILD,n=35),CTD associated PAH plus ILD group(CTD-PAH+ILD,n=16)and CTD control group(n=34).Pulmonary function test parameters,including total lung capacity(TLC % predicted),forced vital capacity(FVC % predicted),forced expiratory volume in the first second(FEV_(1.0)% predicted),FE_(1.0)%/FVC and diffusing capacity of the lung for carbon monoxide(DLco,% predicted)were measured and compared among the four groups.Results One hundred and forteen eases were included and predominantly female with average onset age of 35～39 years old.CTDs that were predisposed to lung diseases were mixed connective disease(MCTD),systemic sclerosis(SSc),systemic lupus erythematosus(SLE)and primary Sj(o)ren syndrome(pSS),in order.There were 10,29 and 46 percent of patients presented with decreased TLC% in CTD-PAH,CTD-ILD and CTD-PAH+ILD group respectively,50,36 and 71 percent of patients with decreased FVC% respectively,54,47 and 71 percent of patients with decreased FEV_(1.0)% respectively,and 100,82 and 100 percent with decreased DLco% respectively.ANOVA analysis demonstrated that TLC%,FVC%,FEV_(1.0)%,DLco% had significant differences between CTD control group and each of the CTD associated lung disease group(P＜0.05),although none of them was lack of difference between the PAH and ILD groups.TLC% was significantly higher in CTD-PAH group than CTD-PAH+ILD group[(89±15)% vs(79±12)%,P＜0.05],while FVC% was significantly lower in CTDPAH+ILD group either than CTD-PAH group or than CTD-ILD group[(81±13)%,(80±16)% vs(65±22)%,P＜0.05].ConclusionPulmonary function test may be valuable in early screening for CTD associated lung disorders than distinguishing CTD-PAH from ILD,which usually reveal restrictive ventilation dysfunction and/or diffusing capacity dysfunction.