The research on intracellular trafficking of adenovirus has been described mainly through observations of subgroup C adenoviruses in transformed cell lines. The basic elements of the trafficking pathway include binding to receptors at the cell surface, internalization by endocytosis, lysis of the endosomal membrane, escape to the cytosol, intracellular trafficking along microtubules, nuclear pore docking, and viral genome translocation into the nucleus. More than 80% of the adenovirus genome is delivered to the nucleus in a highly efficient manner in approximately 1 h. However, exceptions to this trafficking pattern have been noted, including: variations based on target cell type, cell physiology, and adenovirus serotype. This review summarizes mechanism of adenovirus infection pathway and intracellular trafficking, providinging a foundation for the development of clinical adenoviral vector.
Adenoviridae ; physiology ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Cytoplasm ; virology ; Endocytosis ; Endosomes ; virology ; Genetic Vectors ; Humans ; Microtubules ; Virus Internalization

ObjectiveTo investigate the efficiency of autologous transplantation of peripheral blood stem cells for treatment of patients with diabetic lower limb ischemia.MethodsEighteen patients of type 2 diabetes with diabetic lower limb ischemia (30 legs) were treated by autologous transplantation of peripheral blood stem cells.ResultsThe limb pain, cool feeling and numbness feeling improved significantly after PBSC transplantation,the improvement rate were 96.7％, 100.0％ and 95.8％ respectively.Intermittent claudication was also relieved significantly, total remission rate was 76.9％.The ABI and TcPO2 of patients increased significantly at 3 months after transplantation.After the transplantation ABI raised from 0.60 ± 0.11 to 0.71 ±0.12(t =-6.882, P ＜ 0.01) .93.3％ of patients' TcPO2 raised in different degrees.The foot infections were well controlled.Ulcer or toes gangrene got better or healed.No obvious complications or adverse reaction were observed after the transplantation.ConclusionAutologous transplantation of peripheral blood stem cells shows to be a simple, safe and effective method in treating patients with diabetic lower limb ischemia.

[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

Objective To investigate the safety and efficacy of hepatitis B surface antigen (HBsAg) positivity of the donors on graft survival and liver complications in HBsAg (+) renal transplant recipients.Methods We retrospectively evaluated 96 HBsAg (+) patients who received HBsAg(+) donor kidney transplant fellow-up during 20～ 139 months,in order to observe the renal allograft dysfunction,liver dysfunction and others complications.Results All 96 patients underwent renal transplantation successfully in our hospital.during the follow-up period,18 cases accepted entecavir-treated,one case lost graft function,two cases died,one of them developed drug resistance and liver function failure,the other because of cancer of the liver.Twenty-three of the 78 lamivudinetreated patients (29.5％) developed drug resistance in 7～96 months,and 3 cases developed liver function failure,2 cases died and one cured,15 of the 19 cases who been salvage treated with entecavir was successful and well tolerated after 1 year,2 cases who been salvage treated with adefovir and lamivudine with HBV DNA-negative after 12 months and 23 months.The 5-year patient/graft rates of patients who been treated with lamivudine and entecavir were 88.5％/84.6％ and 88.9％/83.3％ respectively.Conclusion It is safe and feasible for renal transplantation from HBsAg(+) donors to HBsAg(+) recipients with antiviral treatment,patients would require lifelong anti-viral suppression and strictly follow-up,which is important for patient and graft survival,anti-viral drugs resistance and the liver complications should be closely monitored and treated.

AIM: To investigate the expression of survivin in pancreas in the streptozotocin-induced diabetic mice. METHODS: Low dose of streptozotocin was used to induce diabetes mellitus in BALB/c mice. Body weight and blood glucose concentrations were examined at 1, 2, 3 and 4 weeks after the streptozotocin injection. Expression of survivin mRNA was detected by real-time FQ-PCR. RESULTS: Survivin was expressed in the pancreas of normal BALB/c mice. Low dose of streptozotocin provoked hyperglycaemia with increased survivin expression in the pancreas, but blood glucose concentration and expression of survivin was not significantly changed in control group. CONCLUSION: Survivin is expressed in the pancreas of normal BALB/c mice. Streptozotocin increases survivin expression in the pancreas, which may be related with islets regeneration.

Objective To explore the distribution of common gene types of thalassemia in Yangjiang, Guangdong, so as to provide a theoretical basis for clinical diagnosis, prevention and treatment. Methods A total of 9, 277 cases were collected from the Department of gynaecologic outpatient, antenatal clinic, outpatient department of Pediatrics and children's health care department from April 2015 to April 2017. PCR + conduction hybridization was used to detect alpha thalassemia and beta thalassemia. Results In 9 277 cases, the detection of α thalassemia in 1 711 cases, accounting for 18. 44％(1 711/9 277); detection of thalassemia in 671 cases, accounting for 7. 24％ (671/9 277), there are 3 kinds of β thalassemia homozygote, α thalassemia and β thalassemia with 100 cases, accounting for 1. 08％ (100/9 277), which detected 16 α thalassemia gene type 14 β thalassemia gene type. Conclusion Guangdong Yangjiang area of αthalassemia by ～(--SEA)/αα and β thalassemia major in β～(CD41-42)/βN and β～(654)/β～N, β～(-28)/β～N, α and β thalassemia gene type compound less investigation for diagnosis, treatment and prevention of thalassemia has important significance, is worth promoting in clinical.

BACKGROUND:As a kind of undifferentiated precursor cells,the phenotypic differentiation of bone marrow mesenchymal stem cells (BMSCs) remains immaturity,thus it presents weak rejection following transplantation.However,the in vitro directional differentiation of BMSCs into neuronal cells is easy affected by various factors.OBJECTIVE:To observe the immunomodulatory effect and the potential of BMSCs differentiate into neuronal-like cells.DESIGN,TIME AND SETTING:A contrast observation was conducted at the Department of Cytology,Third People's Hospital of Wuxi from January 2008 to March 2009.MATERIALS:Bone marrow was harvested from chips of cancellous or ilium bone dudng hip joint surgery.METHODS:Firstly,the BMSCs were separated and cultured to establish mixed lymphocyte reaction (MLR) system.Secondly,2 samples of peripheral blood mononuclear cells (1×10~5/well) were added into 96-well plate,and then BMSCs treated by mitomycin were added according to different ratios (BMSCs/peripheral blood monouclear cells).At the end,the cells were cultured as follows:Method 1:DMEM+10% fetal calf serum+1 μmol/L RA +20 μg/L basic fibroblast growth factor+20 μg/L epidermal growth factor.Method 2:DMEM+2% dimethyl sulfoxide +100 μmol/L butylated hydroxyanisole.MAIN OUTCOME MEASURES:The growth rate of lymphocyte was measured by ~3H-Thymidine,and the effect of BMSCs on lymphocyte proliferation was observed.Additionally,the differentiation potential of BMSCs into neuronel cells was determined by immunofluorescenca and immunohistochemistrical staining.RESULTS:①The BMSCs inhibited lymphocyte proliferation in MLR system and the influence on proliferation of lymphocyte was direct related to ratio of BMSCs.②Under a light microscope,cytoplasm of BMScs were shrinkd,which presented typical perikaryon morphology at hour 2 after culture with method 1.The majority of BMSCs were formed neuronal-like cells without number changes at hours 3-5,which turned to be dipolar or multipolar neuronal shapes at day 3.There were 60%-70% neuronspecific enolase,45%-50% glial fibrillary acidic protein were positive expressed.However,the positive rate of nidogen was decreased 3.4%.Cells cultured with method 2 became smaller after 2 hours,formed dipolar or multipolar body cells,and most of cells were died after 48 hours.The 40%-50% neuronspecific enolase,35%-40% glial fibrillary acidic protein was found to be positive.The positive rate of nidogen was temporary increased to 63% at hour 2 after culture；however,it was decreased to 1.6% after 48 hours.CONCLUSION:BMSCs can differentiate into neuronal-like cells,as well as inhibit lymphocyte proliferation in MLR system,which possess down regulation effect on alloimmunity-reaction.

OBJECTIVE: To determine the clinical effect of standardized training and management of peripherally inserted central catheter (PICC) and catheter-related complications. METHODS: A total of 610 patients were divided into a control group and an observation group, the control group (n=300) were catheterized by trainees who received "short-term intensive training", the observation group (n=310) by "system standardized training and management". The clinical efficacy of catheterization and the rate of catheter-related complications were compared. RESULTS: There was significant difference in the one-time puncture success rate, one-time cannulation success rate, the time for operation and the pain score between the 2 groups (all P<0.01), and there was also significant difference in the occurrence of catheter extrusion, plug, arrhythmia, catheter-related thrombosis, phlebitis, puncture point effusion and catheter-related infection between the 2 groups (all P<0.05). CONCLUSION Standardized PICC training and management can improve the effect of catheterization and reduce the incidence of PICC-related complication.
Catheter-Related Infections ; prevention & control ; Catheterization, Peripheral ; methods ; Humans ; Incidence ; Inservice Training ; Thrombosis

Objective To investigate the clinical value of serum lipoprotein associated phospholipase A 2 (Lp-PLA2)as a biomarker for benign prostatic hyperplasia(BPH).Methods A total of 65 serum samples of BPH patients were selected as BPH group,64 serum samples of healthy male as control group.The serum lev-el of Lp-PLA2,total prostate specific antigen(tPSA)and free prostate specific antigen(fPSA)in both groups were detected,and the specificity and sensitivity of Lp-PLA2,F/T ratio and combine both were analyzed by ROC curve.Results The serum level of Lp-PLA2,tPSA and fPSA in BPH group were significantly higher than those in control group,the different were significant(P<0.05).ROC curve analyze shown that the area under the curve(AUC)of Lp-PLA2 was 0.763,95% confidence interval(CI)was 0.680-0.833;AUC of F/T ratio was 0.715,95% CI 0.633 -0.795;AUC of Lp-PLA2 combined with F/T ratio was 0.832,95% CI 0.756-0.892.Conclusion The serum level of Lp-PLA2 could be used as a potential diagnostic marker for BPH,and the diagnostic value of Lp-PLA2 combined with F/T ratio was better than ther single indicator. Key words:benign prostatic hyperplasia; lipoprotein associated phospholipase A 2; total prostate spe-cific antigen; free prostate specific antigen; F/T ratio

Background::Hepatic ischemia-reperfusion injury (HIRI) remains a common complication during liver transplantation (LT) in patients. As a key downstream effector of the Hippo pathway, Yes-associated protein (YAP) has been reported to be involved in various physiological and pathological processes. However, it remains elusive whether and how YAP may control autophagy activation during ischemia-reperfusion.Methods::Human liver tissues from patients who had undergone LT were obtained to evaluate the correlation between YAP and autophagy activation. Both an in vitro hepatocyte cell line and in vivo liver-specific YAP knockdown mice were used to establish the hepatic ischemia-reperfusion models to determine the role of YAP in the activation of autophagy and the mechanism of regulation. Results::Autophagy was activated in the post-perfusion liver grafts during LT in patients, and the expression of YAP positively correlated with the autophagic level of hepatocytes. Liver-specific knockdown of YAP inhibited hepatocytes autophagy upon hypoxia-reoxygenation and HIRI ( P <0.05). YAP deficiency aggravated HIRI by promoting the apoptosis of hepatocytes both in the in vitro and in vivo models ( P <0.05). Attenuated HIRI by overexpression of YAP was diminished after the inhibition of autophagy with 3-methyladenine. In addition, inhibiting autophagy activation by YAP knockdown exacerbated mitochondrial damage through increasing reactive oxygen species ( P <0.05). Moreover, the regulation of autophagy by YAP during HIRI was mediated by AP1 (c-Jun) N-terminal kinase (JNK) signaling through binding to the transcriptional enhanced associate domain (TEAD). Conclusions::YAP protects against HIRI by inducing autophagy via JNK signaling that suppresses the apoptosis of hepatocytes. Targeting Hippo (YAP)-JNK-autophagy axis may provide a novel strategy for the prevention and treatment of HIRI.