Objective To study the expression of survivin and its relationship with the proliferative activity in hepatocellular carcinoma.Methods Using immunohistochemistry ABC methods, the expression of survivin and PCNA were evaluated in 31 hepatocellular carcinoma, 8 cirrhosis of liver and 4 normal livers tissues.Results No expression of survivin were detected in normal livers and cirrhosis of liver. In contrast, survivin protein was expressed in 19 of 31 patients with HCC(61.3%). When compared with normal livers and cirrhosis of liver, hepatocellular carcinoma had significantly more frequencies of survivin expression(P

ObjectiveTo investigate the value of ultrasonography in the diagnosis of the papillary thyroid microcarcinoma (PTMC) in patients with Hashimoto’s thyroiditis (HT).MethodsThis retrospective study used data from Ruijin Hospital of Shanghai Jiaotong University of Medicine during November 2012 to December 2013. A total of 111 small thyroid nodules (75 PTMC/36 benign nodules) with 107 HT cases which were pathologically conifrmed were included in this study. The sonographic characteristics of nodules were investigated, including nodule aspect ratio, shape, border, margin, acoustic halo, internal structure, echo level, microcalciifcations, rear acoustic attenuation, vascular pattern and extent of the blood supply and the types of thyroid tissue echogenicity. Chi-square test and Fisher′s exact probability method was used to compare the differences of the sonographic characteristics between the benign nodules and malignant nodules. With surgical pathology as the gold standard, computing the value of ultrasonography in the diagnosis of PTMC with HT, including the sensitivity, speciifcity, diagnostic accuracy, positive predictive value and negative predictive value.ResultsA total of 111 thyroid tiny nodules (75 PTMC/36 benign nodules) with 107 HT cases which were pathologically conifrmed were included in this study. The results showed 111 small thyroid nodules as solid hypoechoic. Four indexes between PTMC and benign nodules had statistical signiifcance, such as margin, microcalciifcations, vascular pattern and extent of the blood supply. The other six indexes between PTMC and benign nodules had no statistical significance, such as aspect ratio, shape, border, acoustic halo, rear acoustic attenuation and the types of thyroid tissue echogenicity. Ultrasound diagnostic accuracy of small tyroid nodules in patients with HT was 74.77% (83/111). The sensitivity, specificity, diagnostic accuracy, positive predictive value and negative predictive value of the ultrasound diagnosis of PTMC were 93.33% (70/75), 36.11% (13/36), 74.77% (83/111), 75.27% (70/93), and 72.22% (13/18), respectively.ConclusionsCompared with general population, some classic ultrasound features became less effective in patients with HT. However, ultrasonography has some differential diagnostic value in these cases.

Objective To investigate the effect of rosiglitazone on the CD4+regulatory T cells in the patients with latent autoimmune diabetes in adults(LADA).Methods The CIM+T cells from IADA patients were isolated with anti-CD4-dynal magnetic beads.The expression of Foxp3 mRNA,along with peroxisome proliferators activator receptors gamma(PPARγ)mRNA and TGF-131 mRNA was determined.The effect of rosiglitazone on CD4+T cells was measured,after treated with rosiglitazone for 48 h.Cell viability was assessed by Mtit assay.The proliferation was assayed with 3 H-TdR.Two-color staining(anti-CD4,anti-CD25)flow cytometric analysis was employed to measure the percentage of CD4+CD25+T cells of Deriph eral blood.Resuits PPARγmRNA was expressed in peripheral CD4+T lymphocytes.RosiglitazoBe inhibited phytohemagglutinin(PHA)-induced human CD4+T cell proliferation in dose dependence.The percentage of CD4+CD25+T cells showed no significant change after the peripheral blood culture with 1 μmol/Land 10μmot/L rosiglitazone.10 μmol/L of rosiglitazone induced Foxp3 mRNA expression in vitro (3.27fold,P<0.05),whereas TGF-β1 mRNA expression did not change.Furthermore,only 1 μmol/L of rosiglitazone could promote Foxp3 mRNA expression if adding IL-2(10 U/m1)in cultures(3.48 fold.P

To train medical students' clinical thinking ability is an important aspect of clinical probation practice and also the common goal of dermartology probation training. This article analyzes the problems around the cultivation of clinical thinking ability during probation practice in our department,and proposes the corresponded measures to improve it.

Objective To observe the morphology of embryonic stem cell-derived neurons after transplanted into A?-injured rat hippocampus. Methods Neural precursor cells (NPCs) were generated from mouse embryonic stem cells (ESCs) expressing EGFP with modified serum-free methods and then transplanted into the hippocampus of A?1-40-injuried rats. The morphology, neurotransmitter phenotypes and receptors of EGFP-positive donor cells were observed with immunofluorescene methods. Results The engrafted NPCs survived and differentiated into glutamatergic and GABAergic neurons and expressed both glutamatergic and GABAergic neurotransmitter receptors. Synaptophysin-positive dots were found surrounding somata and dendrites of transplanted neurons, suggesting the presence of presynaptic terminals adjacent to their membranes. Conclusion NPCs derived from ESCs can differentiate into excitatory and inhibitory neurons after grafted into Alzheimer's disease model rats, and maybe form synapse with the host neurons.

Objective · To study the electrophysiological effect of (S)-OTS·HCl on the heart. Methods · The conventional intracellular recording, electrocardiograph (ECG) and Langendorff cardiac perfusion technique were employed to investigate the effect of (S)-OTS·HCl on in-vivo and in-vitro hearts of guinea pigs and rabbits. Results · (S)-OTS·HCl could bind to M2 muscarinic receptors and dose-dependently prolong the RR intervals significantly in vivo. It had no effect on resting potential (RP), action potential amplitude (APA), and maximum upstroke velocity of phase 0 (Vmax) of ventricular myocytes. Instead, 1×10-5 mol/L (S)-OTS·HCl could shorten the action potential duration at 50 percent repolarization (APD50) and APD90 to 91.6% and 90.9%, respectively. And the spontaneous depolarization rate of phase 4 (SDR) of sinus nodes was reduced to its 13.7% when rabbit sinus nodes were exposed to 1×10-7 mol/L (S)-OTS·HCl. (S)-OTS·HCl could inhibit Ca2+channel effectively. It decreased APA and Vmax of sinus nodes and attenuated the cardiac contractility in vitro. Conclusion · (S)-OTS·HCl is a potent cholinergic agonist and has negative chronotropic, dromotropic, and inotropic effects on hearts via binding to M2 muscarinic receptors.

[ABSTRACT]AIM:Toinvestigatetheeffectsofmechanicalstretchontransientoutwardpotassiumcurrent(Ito), inward rectifier potassium current ( IK1 ) and action potential duration ( APD) of cultured neonatal rat atrial myocytes . METHODS:Neonatal rat atrial myocytes were isolated and cultured on silicone sheeting with or without stretch for 24 h. The silicone membrane area was increased by 12%in stretched group.The cells without stretch served as control .Ito, IK1 and APD were recorded by the technique of whole-cell patch clamp.RESULTS:Compared with control group, Ito density in stretched myocytes was significantly reduced [(1.6 ±0.4) pA/pF vs (12.1 ±2.9) pA/pF, P<0.01], whereas IK1 density was increased [(-10.8 ±0.8) pA/pF vs (-8.8 ±0.9) pA/pF, P<0.01].The APDs at 50%and 90%levels of repolarization ( APD50 and APD90 ) in the stretched cells were obviously decreased than those in non-stretched cells [(10.5 ±1.4) ms vs (15.5 ±2.4) ms, (30.0 ±2.8) ms vs (56.3 ±3.6) ms, P<0.01].CONCLUSION: Stretch stimulation leads to the reduction of Ito density, the increase in IK1 density and the shortness of APD in cultured rat atrial neonatal myocytes , which may contribute to atrial electrical remodeling induced by pressure overload .

AIM: To compare the methods of two currently employed isolation methods for endothelial progenitor cells (EPCs): from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133~+ cells, by defining the cell morphology, phenotype, reproductive activities and function in vitro, providing a reference for clinic application. METHODS: PBMCs from the healthy subjects were used for CD133~+ sorting or not. The two groups of isolated cells were suspended in complete medium M199 for 7 d to 14 d. EPCs phenotype were characterized by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and VEGF concentration was measured using an ELISA kit. Matrigel experiment and migration assay were imitated vascularization in vivo. RESULTS: PBMCs produced more colony-forming units (CFU) than CD133~+ cells from the same volume of blood (P<0.01). From 7 d to 14 d, the two groups show decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144~+ cells in CD133~+ group were lower than those in PBMCs groups (P<0.01). Cells in PBMCs group secreted more VEGF than that in CD133~+ group on 7 d (P<0.01). Compared to CD133~+ group, PBMCs group showed more potential of proliferation and vascularization in vitro. CONCLUSION: CD133~+ sorted cells show a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, which is unable to differentiate to mature endothelial cells, indicating that it's not a preferential way to obtain EPCs for clinic therapy.

BACKGROUND:Hematopoietic reconstruction of malignant tumor in hematopoietic system is related to disease itself,pretreatment program and therapeutic tool after transplantation;especially,mobilization.collection and cryopreservation of auto-peripheral blood stem cell play a key role in successful reconstruction of hematopoietic system after transplantation.OBJECTIVE:To investigate the reconstruction of hematopoietic system through mobilization, collection and cryopreservation of auto-peripheral blood stem cell in patients with malignant tumor and analyze the effective factors on quantity and quality of auto-peripheral blood stem cell.DESIGN:Case analysis based on malignant tumor in hematopoietic system.SETTING:Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA;Department of Blood,Zhujiang Hospital,Nanfang Medical University.PARTICIPANTS:A total of 18 patients with malignant tumor in hematopoietic system were selected from Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.Their ages ranged from 1 6to 56 years.Among them,2 patients had acute myelogenous leukemia(AML),1 acute lymphoblastic Ieukemia(ALL),2 lymphoblastic Ieukemia (LL),2 chronic granulocytic leukemia(CGL),4 multiple myeloma(MM),and 7 non.Hodgkin lymphoma.Granulocyte colony-stimulating factor(G-CSF)was made by Chugai Pharmaceutical Company Limited (batch number:N3G31).METHODS:①All patients were mobilized with associated chemotherapy+G-CSF.Associate chemotherapy:Patients with leukemia were given 2 g/m2 arabinosyl cytosine every 12 hours from lhe first to the third days and 200 mg/m2 etoposide or 50 mg/m2 fludarabine from the first to the fifth days. In addition patients with MM were treated with arabinosyl cytosine as the same way mentioned above and with 1 g/m2 cyclophosphamide from the first to the second days. And patients with lymphoma were given 2 g/m2 cyclophosphamide from the first to the second days. When numbers of leucocyte of all patients decreased below 1.0×109L-1 after chemotherapy.G-CSF started mobilization and the collection was stopped with 5μg/(kg·d)subcutaneous injection.②When numbers of leucocyte increased to (4.0-10.0)×109 L-1,hemopoietic stem cells of peripheral blood were collected till the amount of mononuclear cells≥4.0×108/kq or numbers of CD34+ cells≥2.0×108/kg.And then,the samples were dealt with cooling device.maintained in liquid nitrogen at-196℃ and defrosted in water bath at 37-40℃.③Focal sites of patients were pretreated with local irradiation with 200 cGy/time and 5 times/week for 4 successive weeks.The total dosage was 40 Gy.At 48 hours later,(55.3±28.7)mL hemopoietic stem cells of peripheral blood were transfused back. And the duration from transfusion to collection was about(56.5±22.3)days.300 μg/d G-CSF was subcutaneously injected into all patients at 1 day after transplantation and the reaction was stopped at the phase of neutrophil≥0.5×109L-1. Finally. Refusing-staining rate of trypan blue of peripheral blood stem cell, amount of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were detected before and after thaw.MAIN OUTCOME MEASURES:①Collection of auto-peripheral blood stem cell;②survival rate and related markers of auto-peripheral blood stem cell after cryopreservation;③hematopoietic reconstruction of auto-peripheral blood stem cell after transplantation.RESULTS:All 18 patients with malignant tumor in hematopoietic system were involved in the final analysis.The mean collection time of auto-peripheral blood stern cell was 12.6 days after chemotherapy.the collection times were 1.9.total number of leucocyte was(8.93±1.27)×1 0.L-1 on the first day,and collection rate of mononuclear cell was (138.33±28.61)%. ②Refusing-staining rate of trypan blue of auto-peripheral blood stem cell was similar before and after cryopreservation[(96.26±1.33)%, (92.75±2.04)%,P＞0.05].in addition,after cryopreservation,recovery rates of mononuclear cells,CD34+ cells and granulation-monophyly progenitor cell were(91.96±1.37)%, (85.94±0.64)%and (87.69±4.53)%,respectively.Collection rate of mononuclear cells,number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were lower in patients with myeloma than in those with leukemia and lymphoma (t=2.524-3.268.P＜0.05).③At 15 days after transplantation,15 patients had the neutrophil≥0.5× 109L-1;at 20 days after transplantation,blood platelet was≥20 × 100 L-1.granulation-monophyly progenitor cells[(18.67-26.82)× 105/kg] of 5 patients grew poorly if the course of chemotherapy was more than 10 times.Among them,3 patients had delayed hematopoietic reconstruction after transplantation of auto-peripheral blood stem cell.CONCLUSION:①High-dose chemotherapy combined with G-CSF can shorten collection time of peripheral blood stem cell and improve collection rate of mononuclear cells.②Increase of chemotherapy times before transplantation can affect quantity and quality of auto-peripheral blood stem cell and cause delayed hematopoietic reconstruction.

Objective To investigate the incidence and risk factors of surgical site infection(SSI)in neurosurgical patients in a tertiary first-class hospital,and provide reference for the prevention and control of SSI.Methods 47 neurological patients with SSI (49 patients developed SSI,2 were excluded from study due to the lack of appropriate control subject)from December 31 ,2011 to December 31 ,2012 were as infected group,and 94 patients without SSI (1 ∶2 matching)were as non-infected group,risk factors for SSI were analyzed retrospectively.Results There was no significant difference in general condition of two groups of patients (all P >0.05 );among 3 708 patients,49 (1 .32%)developed SSI;intracranial infection was the main type of SSI (89.80%);27 patients were performed ce-rebrospinal fluid (CSF)bacteriological detection,6 (22.22%)of whom were positive for CSF bacteriological detec-tion.Univariate conditional logistic regression analysis showed that risk factors for SSI in neurosurgical patients were operational risk assessment score (OR =2.04),frequency of preoperative antimicrobial use(OR =3.15 ),fre-quency of intraoperative antimicrobial use(OR=2.58),duration of operation(OR=2.70),surgical blood loss(OR=1 .72),indwelling drainage tube(OR=4.30),duration of indwelling drainage tube after operation(OR=2.06),and time for initial dressing change(OR=1 .66);Multivariate conditional logistic regression analysis showed that the in-dependent risk factors for SSI were frequency of preoperative antimicrobial use(P =0.03,OR =4.86),duration of operation(P =0.05,OR = 2.89 ),and time for initial dressing change after operation (P = 0.01 ,OR = 1 .92 ). Conclusion Risk factors for SSI in department of neurosurgery are multiple,duration of operation,duration of in-dwelling drainage tube after operation,and time for initial dressing change after operation are major risk factors.