In order to investigate the effect of anti-hepato-cellular carcinoma of HSV-tk suicide gene system, we constructed the HSV - tk recombinant retroviral vector pLXT. SMMC - 7721 hepatocellular carconoma cells more transfected with pLXT by lipofectin were obtained by subsequent G418 screen. 3H-TdR incorparation assay showed that HSV - tk/ACV had strong cytotoxic effect on HSV - tk gene transfected tumor cells. Lipofectin pLXT complex was directly injected into murine H22 hepatoma tissue, followed by delivery of ACV prodrup, and it was found that the tumor growth masses more greatly reduced. Animals treated with Liptk + ACV and tk + ACV had an apparent reduction of tumor size as compared with the animals in other six groups ( P

BACKGROUND:Articular cartilage is a highly diferentiated tissue, which is very limited in its ability to repair after injury. Stem cel therapy for cartilage repair has completely solved this problem. OBJECTIVE: To investigate the mechanism that diferent growth factors induce the diferentiation of bone marrow mesenchymal stem cels into chondrocytes. METHODS:Passage 5 rat bone marrow mesenchymal stem cels were induced by diferent growth factors and their combinations, including transforming growth factor beta 1 (TGF-β1) group, TGF-β1+insulin growth factor-1 (IGF-1) group, and bone morphogenetic protein 2 (BMP-2)+IGF-1 group, TGF-β1+BMP-2 group, TGF-β1+IGF-1+ BMP-2 group, and blank control group. At 21 days of induction, cels were stained with alcian blue and alizarin red; RT-PCR was employed to detect colagen I mRNA expression. RESULTS AND CONCLUSION:Alcian blue staining showed metachromasia in the cytoplasm and mesenchyma, and proteoglycan was expressed green; alizarin red staining showed no orange calcium nodules. The bone marrow mesenchymal stem cels were preliminarily deduced to diferentiate into chondrocytes, but could not express the cel phenotypes of bone cels. In the blank control group, the expression of colagen I mRNA was negative. Compared with the TGF-β1 group, the mRNA expression of colagen I was lower in the BMP-2+IGF-1 group, but higher in the TGF-β1+BMP-2 group and TGF-β1+IGF-1+BMP-2 group (P < 0.05). Moreover, the expression of colagen I mRNA was highest in the TGF-β1+IGF-1+BMP-2 group (P < 0.05). These findings indicate that TGF-β1 alone is able to induce the chondrogenic diferentiation of bone marrow mesenchymal stem cels, and the combination of TGF-β1, IGF-1 and BMP-2 can play the biggest role to induce the chondrogenic diferentiation of bone marrow mesenchymal stem cels.

Objective To study the role of schizophrenia health education in intervening relapse of schizophrenia. Method 78 patients with schizaphrenia have been intervened by useing SHE about their staying in hospital and conditions out of hospitals after having recovered for three years, then compared with 85 schizophrenics receiving no SHE. Result It was clear that general rate of relapse in the study group was significantly lower than that of the control group. The patients obey medication and recovery of social function in the study group were significanly better than that in the controlgroup. Conclusion The SHE plays an important role in intervening relapse of schizophrenia and improving complete rehabilitation of schizophrenic.

Objective To explore the method of primary isolation, cultivation and identification of rat brain microvessel pericytes. Methods The brain tissue of 10 3 week-old Wistar rats was separated sterilely. The brain microvessel fragments were separated using two-step enzyme digestion and one-step gradient centrifugation and were seeded in 35-mm dishes for primary culture. The cell morphology was observed by phase contrast microscopy; the immunofluorescence assay was used to identify the associated antigns, such as the α-smooth muscle actin (α-SMA), neuron-glial antigen 2 (NG2), von Willebrand factor (vWF), and glial fibrillary acidic protein (GFAP). Methyl thiazolyl tetrazolium was used to determine the cell growth curve. Results Pericytes climbed out from the adherent brain microvascular fragments around,showing polygonal, and the cell fusion was 95％ after 12-14 days. Immunofluorescence staining revealed that the molecular markers of the pericytes α-SMA and NG2 related antigens showed double positive, while the vWF and GFAP related antigens showed double negative and the cultured cells were confirmed as brain microvascular pericytes. The growth rate of primary cells was slower. The passage cells entered into logarithmic growth phase after 36 to 60 hours and entered into plateau phase after 72 to 108 hours. Conclusions This method may successfully isolate rat brain microvascular pericytes with higher purity.

Objective To observe the dynamic changes of leptin and transforming growth factor (TGF)-β1 in formation of liver fibrosis in rats,and the role of leptin in liver fibrosis.Methods Six rats were served as normal control(0 week)and killed at the beginning of the study.Thirty-two rats were subcutaneously injected with 60%CCl4 to establish fibrotie models.The rats were than sacrificed at 3,6,9 and 12 weeks with eight each.Expressions of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βRⅡmRNA in liver were detected by reverse-transcription polymerase chain reaction(RT-PCR).Expression of Smad3 and Smad7 proteins were determined by Western blot.The expression and loealization of leptin,Ob-Rb and TGF-β1 in liver tissue were detected by immunohistochemistry.Results The expression of leptin mRNA,Ob-Rb mRNA,TGF-β1 mRNA and TGF-βR Ⅱ mRNA in 0 week were 0.43±0.45,0.57±0.21,0.19±0.12 and 0.30±0.22,respectively,and increased to 1.27±0.19,1.70±0.29,1.70±1.00 and 2.14±1.02 at 12 weeks(P＜0.05).At the same times the level of Smad3 increased from 0.44±0.24 at 0 week to 1.75±1.05 at 12 weeks.While the expression of Smad7 was decreased from 1.35±0.12 at 0 week to 0.74±1.21 at 12 weeks(P＜0.05).The expressions of leptin,Ob-Rb and TGF-β1 were increased with the formation of the fibrosis(P＜0.01).Conclusions With the development of liver fibrosis,the expressions of leptin,Ob-Rb mRNA and protein are increased and improved the formation of the fibrosis.Its mechanisms may be correlated with up-regulation of TGF-β1,TGF-βR Ⅱ and Smda3 expressions and down-regulaton of Smad7 expression by leptin.

Objective:To identify the role of transcription factors Sp1 and Sp3 in the expressional regulation of ezrin in human esophageal carcinoma cells.Methods:Esophageal carcinoma EC109 cells were transfected with expressing vectors CMV-Sp1 or CMV-Sp3,and the effect of Sp1 and Sp3 over-expression on ezrin mRNA and protein expression was determined by real time RT-PCR and Western blot analysis.Furthermore,EC109 cells were cotransfected with the ezrin promoter-directed luciferase reporter vector and control vector pRL-TK along with transcription factor expression vector.The roles of Sp1 and Sp3 in ezrin promoter activation and whether this activation occurred through the Sp1 binding site,-75/-69,were analyzed by dual-luciferase reporter assay system.Results:Over-expression of transcription factors Sp1 and Sp3 significantly increased the expression of ezrin mRNA and protein and the ezrin promoter activity in EC109 cells.Sp1 and Sp3 enhanced the promoter activity through different binding sites and only Sp1 did that through the-75/-69 site.Conclusion:Sp1 and Sp3 can regulate ezrin expression in EC109 cells.

0.05, while the differences between 2 groups in cooling effects on the points of cooling 2.5 hours and 4.0 hours were significant, F

A 19-year-old female presented with a dark erythematous maculopapule measuring 2 cm in diameter on the left lower limb for more than one year.Physical examination revealed an enlarged cherry-like lymph node in the right inguinal region.Histopathology of the lesion revealed that the normal structure of skin disappeared absolutely,and there was a diffuse infiltrate of medium-sized lymphoid tumor cells with unclear nucleoli.Immunohistochemically,the tumor cells strongly expressed CD4,CD56 and CD123,and partially expressed terminal deoxynucleotidyl transferase (TdT).Flow cytometric analysis of bone marrow aspirates showed that abnormal cells amounted to 59.9％,and were positive for CD123st (strong),human leucocyte antigen-DRst (strong),CD56,CD304 (blood dendritic cell antigen-4),CD7,CD11b,CD33,CD36 and chemokine (C-X-C motif) receptor 4,CD13 (dim),CD4 (partial) and CD117 (partial).Transmission electron microscopy of bone marrow cells revealed that there were massive uniformly sized lymphoid cells with thick processes on cell surfaces,a high nucleus/cytoplasm ratio,centrally located round nuclei occasionally with deep gyrus-like notches,clumped marginated heterochromatin and multiple nucleoli.Moreover,a small amount of cytoplasm was observed in the lymphoid cells with the presence of mitochondria and endoplasmic reticulum arranged in concentric circles,which were characteristic of plasma cells.The features of both dendritic cells and plasma cells were found in the tumor cells through transmission electron microscopy.Based on the above findings,a diagnosis of blastic plasmacytoid dendritic cell neoplasm (BPDCN) was made.

Objective To explore the protective effects of quercetin on damage induced by oxidative stress and to clari?fy its molecular mechanism. Methods Chang liver cell cultures were randomly divided into control groups, H2O2 group and 3 doses of quercetin groups. Cell survival rate was detected with MTT. Cell apoptotic rate was measured by FACS(Fluores?cence-activated cell sorting). Intracellular reactive oxygen species (ROS) level in Chang liver cells were tested by flow cy?tometer. The DCF fluorescence intensity of DCFH-DA-stained intracellular ROS was observed by fluorescence microscope. The levels of malondialdehyde (MDA), superoxide dismutase(SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were determined in liver cells using commercial available kits. The expression of Nrf2 were detected by Western blot. Re?sults Compared with control, cell survival rate and levels of SOD, CAT and GSH-Px decreased significantly in H2O2 group (P < 0.05 ),while cell appotosis rate, content of MDA and mean fluorescence intensity(MFI) increased in H2O2 group (P <0.05). In comparison with H2O2, expression of Nrf2 protein was higher in all three quercetin treatment groups (P<0.05). Con?clusion Quercetin protected Chang liver cells from H2O2-induced oxidative stress, which may be caused by the increased ex?pressions of down stream antioxidant genes via activating the Nrf2-ARE signaling pathway.

Objective:To investigate the long-term toxicity of recombinant human interleukin-11(rhIL-11) in cynomolgus. Methods: Eighteen cynomolgus were randomized into 4 groups: control group(2/sex), low dose group(2/sex), medium dose group(2/sex)， and high dose group(3/sex). The drug groups were sc adminstered 0.1, 0.3 and 1.0 mg/kg of rhIL-11 for 90 days with a 30-day recovery period. The clinical signs were observed, electrocardiogram, hematological, biochemical, urinary and immunological parameters were measured, organ masses were weighed, bone marrow and pathological histology were observed. Results: The food consumption, body mass of the drug groups were decreased, the body temperature was increased transiently. One of the low dose group showed restricted movements and tremors. One of the high dose group vomited and another died. Reduced red blood cell(RBC) count, hemoglobin(Hb) concentration, hematocrit(Hct), mean corpuscular volume(MCV), mean corpuscular hemoglobin(MCH), and mean corpuscular hemoglobin concentration(MCHC), dose-related increase of platelet(Plat) counts were present in drug groups. Biochemical examinations revealed dose-related decreases in serum alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH), total proteins(TP) and albumin(Alb) increases in serum alkaline phosphatase(ALP) levels. Positive antibody responses were seen and circulatory immune complex(CIC) was significantly increased in all drug groups. Hypertropy of marrow megakaryocyocytes was noted in the medium and high dose groups. The heart and liver masses were slightly increased in all treatment groups. Treatment-related microscopic findings included dose-related degeneration in the liver and the kidney. The adverse effects were reversed by the end of the recovery period. Conclusion: The target organs and systems are blood, liver, kidney, immmue system and bone marrow. The toxicity injuries were reversible and the no-toxic-effect level is 0.1 mg/kg.