Animals ; Critical Illness ; Enterovirus A, Human ; pathogenicity ; Enterovirus Infections ; diagnosis ; therapy ; Hand, Foot and Mouth Disease ; diagnosis ; therapy ; virology ; Humans

The cerebral ischemia was produced by Pulsinellis method in Sparaque-Dawley rats. The brain edema and survival rate of rats with bilateral carotid and vertebral arteries occlusion for 60 min were observed in ip tetradrine at doses of 1 ~ 4 mg/kg groups and control rats.Superoxide dismutase and malondialdehyde in brain tissue were also measured by pyrogallol method and fluorescence spec-trometry. The results suggested that tetradrine have protective effect on cerebral ischemia, which was related to the inhibition of lipoxide and scavenging of oxygen free radical.

Hepatitis B virus X protein(HBx) has a wide range of biological effects.It not only play important roles in the replication of the virus,but also in infects host cell proliferation and apoptosis,as well as the incidence of hepatocellular carcinoma.HBx trans-activation has a broad regulatory role in the virus itself and host genes.HBx,by interfering with multiple signaling pathways,affected a variety of biological characteristics of the host cell.

Objective To investigate the effects of methylthioadenosine phosphorylase (MTAP) on invasion and migration in breast cancer cells.Methods Human breast cancer cell line MCF-7 cells were treated with MTAPtargeted siRNA to diminish MTAP mRNA.MCF-7 cells proliferation was evaluated by cell counting kit-8,the analysis of cells invasion and migration was performed using Transwell chamber.The expressions of MTAP and matrix metalloproteinase 1 (MMP1) in cell extracts were detected by Western blotting.The experimental divided into blank contrd group,negative control group,MTAP-siRNA experimental group.Results The MCF-7 cells growth was promoted after knockdown the MTAP mRNA.MATP-siRNA experimental group 450 nm absorbance values at 24h,48 hand72 h of the control group were (112.3±11.9)％,(144.4±8.4)％,(169.3±9.4)％ respectively.Cell invasion analysis by Transwell chamber showed 570 nm absorbance values were 0.49 ± 0.06 (control),0.45 ± 0.07 (negative control) and 0.87 ± 0.07 (MTAP-siRNA) respectively.Cell migration analysis by Transwell chamber showed 570 nm absorbance values were 0.46 ± 0.06 (control),0.49 ± 0.08 (negative control)and 0.75 ± 0.07 (MTAP-siRNA) respectively.The expression of MMP1 in MCF-7 cells was upregulated after knockdown the MTAP mRNA.Conclusion The knockdown of MTAP in MCF-7 cell can increase the cells invasion and migration,and this may involve the the MMP1.

In order to observe the replication and expression of HGV RNA genome in HepG2 cells and establish a cell model of HGV infection, HGV RNA genome was prepared in vitro and transfected HepG2 cells with lipofec-tamin. HGV RNA-positive supernatants were used to infect fresh HepG2 cells. RT-PCR, immunohistochemistry and Western blot assays were carried out to detect the replication and expression of HGV in HepG2 cells. Both positive and negative strands of HGV RNA could be detectable in cell culture supernatants and cells at 24h post-transfection. During the culture periods of 90 days, the cells were maintained by changing the medium every 3 or 5 days, and cultured for more than 20 passages. Both strands of HGV could be detectable in culture supernatants and cells. Immunohistochemistry and Western blot results also confirmed that HGV E2 protein could be expressed in the infected HepG2 cells. HGV RNA could also be detectable in the frozen-thawed HepG2 cells infected with HGV RNA genome. Therefore, HGV RNA genome can replicate and express in HepG2 cells, this HGV RNA genome transfected cells model could be used as a cell model in the studies of replication and infection of HGV.

Objective To report a case of psoriasis vulgaris associated with acute myelogenous leukemia(AML)(type M4EO).Methods Clinical data from the patient were collected.Histopathologic examination,and examination of bone marrow and peripheral blood smear were performed.The immunologic types of bone marrow cells were analyzed with FACS.Chromosome and G-banding analyses were carried out with cultured bone marrow cells.Results A33-year-old woman had a history of chronic plaque psoriasis for20years.Her cousin had the same disease history.The patient was treated with various therapeutic regi-mens,most of which were traditional Chinese medicines.Recently the patient suffered from myalgia and chest bone pain,periodic bleeding on gums,fever and so on.The abnormal infantile monocytes and promye-locytes were found with bone marrow smear,and crassitude basophilic granules were noticed in eosinophils.The diagnosis of acute myelogenous leukemia type M4EO was made.The diagnosis was confirmed with the immunologic analysis of born marrow cells with FACS.Chromosome and G-banding analyses revealed her karyotype of46,XX,inv(16)/47,XX,inv(16),+8(2/22).The plaque lesions of psoriasis were regressed after allogeneic bone marrow transplantation and the symptoms of AML were resolved.Conclusion It is the first case report of psoriasis vulgaris associated with acute myelogenous leukemia M4EO which responded to allogeneic bone marrow transplantation.

Objective To evaluate the specificity,sensitivity and accuracy of miniature probe combined with radial scanning endoscopic ultrasonography(EUS)in preoperative TN staging of rectal cancer,and to assess its value in the choice of therapeutic strategy.Methods A total of 60 patients with rectal cancer received EUS assessment before surgery.Diagnosis was made according to TNM standard and compared with those of MRI and postoperative pathological examination.The reference value of EUS for therapy selection was studied.Results According to EUS staging,there were 4 cases of TI,18 T2,30 T3 and 8 T4,among which 7 cases were over-staged and 4 others were under-staged.MRI staging showed 1 case of T1,18 T2,30 T3 and 10 T4,among which 14 were over-staged and 3 others were under-staged.The total accuracy of EUS in T staging and N staging was 81.67%(49/60)and 78.33%,respectively,with the sensitivity and specificity at 71.43%and 91.03%,respectively.Accuracy of MRI for T staging and N staging were 71.67%(43/60)and 83.33%,respectively,with the sensitivity and specificity as 85.71%and 86.96%.Conclusion EUS with combination of miniature probe and radial scanning is effective in preoperative TN staging of rectal cancer with easy manipulation and less pain.

Objective To assess the role of endoscopic ultrasound guided fine needle aspiration (EUS-FNA)for diagnosis of gastrointestinal tract lesions.Methods Sixty-eight patients underwent endoscopic uhrasonagraphy and EUS-FNA between May 2007 and Dec.2008.The result of cytology and/or pathology was compared with that of surgical finding and follow-up study.Results EUS-FNA was successfully performed on 62 patients with lesiorm in oesophagus(4 eases),stomach(19 case),rectum(19 cases),liver(3 case),mediastinum(4 cases)and lymph node(13 cases),and had a successful rate of 91.18%(62/68).Among them,the lesions in 40 patients(64.52%)were cytologically confirmed.The lesions in 10 out of 22 patients(35.48%)were pathologically confirmed.Compared with the results of pathology and long-term followe-up study,the sensitivity,specificity and positive predictive value of EUSFNA in diagnosis of lesions in gastrointestinal tract was 85.48%,100.00%and 90.91%,respectively,The positive predictive value and negative predictive value was100.00%and 57.14%,respectively.Conclusion EUS-FNA is a safe,effective and accurate method for diagnosis of lesions in gastrointestinal tract,and has an important role in cytological diagnosis.

Objective To explore the value of EUS in the diagnosis,differential diagnosis and follow-up of primary gastric lymphoma(PGL).Methods A total of 26 patients whose endoscopic findings were highly suspected for PGL were enrolled in this study.Initial endoscopic examination anti routine biopsy were performed in all patients.EUS were undergone in all cases and its guided large piecemeal biopsy was administered in selected patients.The results of EUS,routine biopsy and routine biopsy combined with a large piecemeal biopsy were compared.Results Compared with biopsy and histopathological results,EUS made the fight diagnosis in 23 cases with overall accuracy of 88.46%,which was significantly superior to routine endoscopy and biopsy(50.0%,P=0.006),but was slightly lower than that of routine biopsy combined with large piecemeal biopsy(88.5%vs.92.3%,P=1.000).EUS can accurately differentiate gastric malignancy from benign disease with accuracy of 100.0%(23/23).Meanwhile EUS accuracy of tumor invasion(T stage)was 100.0%(12/12)as well.Conclusion EUS is valuable in the diagnosis,differential diagnosis and follow-up of PGL.

Objective To investigate the feasibility of inhibiting Galα (1,3)-Gal expression in mouse vascular endothelial cells by lentivirus-mediated RNAi.Methods The shRNA specified to α1,3-GT mRNA was designed and synthesized in vitro and cloned into the lentivirus vector.EOMA cells were infected by recombinant lentivirus.Real-time RT-PCR was used to detect mRNA transcriptional levels of αl,3-GT as well as immunofluorescence and flow cytometry were applied to detect Galα(1,3)-Gal antigen level after gene transfection.Co-culture of infected EOMA and serum of human was done and the survival rate was measured by MTT.Results The αl,3-GT shRNA sequences were cloned into the recombinant lentivirus vector correctly and the lentivirus was produced successfully.The transfection efficiency to EOMA was 75 %.Real-time PCR revealed that the mRNA transcription of α1,3-GT was obviously inhibited by α1,3-GT shRNA recombinant lentivirus with the rate of 88 % (P＜0.05),while there were no obvious differences among control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＞ 0.05).Immunofluorescence and flow cytometry demonstrated the same results that Galα(1,3)-Gal antigen expression in EOMA transfected by α1,3-GT shRNA lentivirus was less than that of control group,no shRNA lentivirus group and negative-shRNA lentivirus group (P＜0.05),but there were no obvious differences among the later three groups (P＞0.05).After co-culture with serum of human,MTT showed the survival rate of EOMA infected by α1,3-GT shRNA lentivirus was obviously increased (P＜ 0.05).Conclusion Recombinant α2,3-GT shRNA 1entivirus is constructed successfully,which can inhibit the expression of α1,3-GT and Galα1,3-Gal in EOMA by RNAi and control hyperacute rejection in vitro.