Tegument protein VP22 is encoded by Pseudorabies Virus (PRV) UL49. To identify the nuclear localization signals of UL49, it is necessary to determine the transport mechanism and biological functions of the VP22 protein. In this study, we identified two nuclear localization signals from UL49, NLS1 (5RKTRVA ADETASGARRR21) and NLS2 (241PGRKGKV247). The functional nuclear localization signal (NLS) of UL49 was identified by constructing truncated or site-specific UL49 mutants. The deletion of both NLS1 and NLS2 abrogated UL49 nuclear accumulation, whereas the deletion of NLS1 or NLS2 did not. Therefore, both NLS1 and NLS2 are critical for the nuclear localization of UL49. And our resuts showed that NLS2 is more important in this regard.
Animals ; COS Cells ; Cell Nucleus ; metabolism ; virology ; Cercopithecus aethiops ; Herpesvirus 1, Suid ; chemistry ; genetics ; metabolism ; Humans ; Nuclear Localization Signals ; Protein Transport ; Pseudorabies ; metabolism ; virology ; Viral Structural Proteins ; chemistry ; genetics ; metabolism

Child, Preschool ; Echocardiography, Transesophageal ; methods ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Male

Objective To investigate the correlation between the formation of pigmented biliary calculus and biliary H.pylori infection.Methods Bile from 35 patients with pigmented biliary calculus and 10 healthy controls was cultured for aerobic,anaerobic and H.pylori.The expression of H.pylori- DNA in bile,bile duct mucosa and pigmented calculus were determined by PCR.The expression of H. pylori associated protein in bile duct mucosa was determined by Western-blot and Warthin-Starry staining.Results H.pylori culture was negative in all bile samples.In 35 patients with biliary pigmen- ted calculus,H.pylori was detected by PCR in the center of calculus,bile and bile duct mucosa of 14.29%,31.43% and 56.67% patients,respectively.Among H.pylori-DNA positive bile samples,7 contained anti-CagA antibodies,and 6 contained Vac A.in addition to Vacuolating cytotoxin(35000), glycoprotein(30000),Urase Band Urase A.Bacteria resembling H.pylori by Warthin-Starry stainning were found in 7 of 30(23.33%)bile duct mueosal samples from patients with biliary pigmented calculus. H.pylori-DNA and its associated protein were not detected in all bile and bile duct mucosae samples from the healthy controls.Conclusions The evidence of H.pylori-DNA and its associated protein in biliary system might indicate the role of H.pylori in the formation of biliary pigmented calculus.

Adult ; Endocarditis, Bacterial ; complications ; diagnosis ; Humans ; Male ; Myocardial Infarction ; diagnosis ; etiology

Objective To study the time distribution of non-agriculture employed population spent in a working day in China, and to provide basic information for intervention strategies.Methods The data of 2002 China Nutrition and Health Survey were used. The information on daily activities including occupation,transportation,exercise,housework,sedentary activity and sleep was described.Results Non-agriculture employed population spent 8.41 h,0.58 h,0.09 h,3.11 h,1.40 h and 7.89 h on occupation, transportation,exercise,sedentary activity,housework and sleep,respectively.Administrator,technologist and clerks spent less time on occupation activity than service workers,production and transportation workers and others did,and they spent more time on sedentary activity.Male spent more time on occupation activity and less time on housework and sleep than female did.People in rural area spent more time on occupation activity than those in urban area,but less time On transportation and sedentary activity.Conclusion Differences in time use among different employed groups,gender and area were found,which should be considered when intervention measure is developed.

Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

To investigate the influence of the difference enhancers on the transport mechanism of chlorogenic acid (CGA) across Caco-2 cells model, a RP-HPLC method was adopted to detect the concentrations of CGA. At the concentrations of 20 to 80 microg x mL(-1), the difference of absorption rate constants (K(a)) was not statistically significant. At the concentrations of 40 and 20 microg x mL(-1), the ratios of apparent permeability coefficients (P(app)) of the apical to basolateral and the basolateral to apical were 1.14 and 1.18, respectively. With the effect of enhancers K(a) and P(app) increased, the absorption half-life (T1/2) decreased. CGA passed through the Caco-2 cell membrane mainly by passive transport. It showed that monocarboxylic acid transporter (MCT) could be involved in the across membrane transport process of CGA. Borneol had no effect on the cell membrane transport processes. The order of increasing absorption of CGA caused by the enhancers was sodium lauryl sulphate > sodium taurocholate > carbomer.
Absorption ; Acrylic Resins ; pharmacology ; Caco-2 Cells ; Cell Membrane Permeability ; drug effects ; Chlorogenic Acid ; pharmacokinetics ; Humans ; Sodium Dodecyl Sulfate ; pharmacology ; Taurocholic Acid ; pharmacology

Adult ; CD56 Antigen ; metabolism ; Carcinoma, Papillary ; diagnosis ; metabolism ; pathology ; surgery ; Endoscopic Ultrasound-Guided Fine Needle Aspiration ; methods ; Female ; Humans ; Male ; Neprilysin ; metabolism ; Pancreatic Neoplasms ; diagnosis ; metabolism ; pathology ; surgery ; Receptors, Progesterone ; metabolism ; Vimentin ; metabolism ; Young Adult ; beta Catenin ; metabolism

OBJECTIVETo investigate the Toxoplasma gondii (TOX) infection in males with sterility and the effect of the infection on the reproductive function of males.

METHODSEnzyme linked immunoabsorbent assay (ELISA) was used to detect TOX-CAg, TOX-IgG and TOX-IgM in the peripheral blood of male patients with sterility.

RESULTSAmong 100 cases of male sterility, 7 were TOX-IgG positive (7%), 16 TOX-IgM positive (16%) and 13 TOX-CAg positive (13%). Among 100 normal males, 7 were TOX-IgG positive (7%), 3 TOX-IgM positive (3%) and 1 TOX-CAg positive (1%).

CONCLUSIONTOX infection may affect the fertility of males and cause male sterility. For this reason, males should prevent themselves from TOX infection.

Adult ; Animals ; Antibodies, Protozoan ; blood ; Antigens, Protozoan ; blood ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Infertility, Male ; epidemiology ; parasitology ; Male ; Toxoplasma ; immunology ; Toxoplasmosis ; complications ; epidemiology

Adolescent ; Cardiac Catheterization ; methods ; Child, Preschool ; Echocardiography, Transesophageal ; methods ; Female ; Heart Septal Defects, Atrial ; diagnostic imaging ; therapy ; Humans ; Treatment Outcome