Aim To express hepatitis C virus glycoprotein E (gE) deleting carboxy-terminal 31 amino acids, and detect anti-E antibody in HCV patients using expressed gE. Methods E gene derived from HCV H strain was inserted into baculovirus transfer vector containing a polyhedrin promotor. The recombinant plasmid was cotransfected into insect cell sf9 with a viral expression vector. The expression of gE was analyzed with Western blot, and the cells were used for dectecting antibodies against E1 and E2 in 35 hepatitis C patients by indirect immunofluorescence. Results A series of proteins with different relative molecular masses(Mr) could be detected by Western blot. Results from indirect immunofluorescence staining showed and only 4 patients were anti-E antibody positive gE was located in cytoplasm. Conclusion HCV gE is expressed successfully in insect cells, the study lay the foundation for further developing HCV vaccine.

AIM:To study alterations of cardiac sarcoplasmic reticulum (SR) function in vitamin D 3-induced calcium overload rats. METHODS: The Ca-overload rat models were prepared by vitamin D 3 plus nicotine. Cardiac SR was seperated by centrifuging. The measurement of SR Ca 2+ uptake and Ca 2+ release activities were preformed by the Millimore filtration technique. Specific SR [ 3H]-ryanodine binding capacity was measured by radioligand method. RESULTS: Compared with control,myocardial calcium content in calcium overload rats increased by 78%( P

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

Objective: To observe the inhibitory effect of scopolamine(Spm) and chlopromazine (Clo) on withdrawal syndromes in morphine dependent rats. Methods: The intensity of withdrawal syndromes on the model of morphine dependent rats was recorded after single or muiltiple subcutaneous administration(sc) of Spm and Clo at different doses. Results: Withdrawal syndromes were markedly decreased when single Spm 1 mg/kg and Clo 0.5 mg/kg combined with morphine were injected (P<0.05). Spm+Clo(sc) had much stronger effects on inhibiting withdrawal syndromes after intraperitoneal (ip) naloxone in morphine dependent rats (P<0.01). Conclusion: Spm can act on Ach-receptor and relieve morphine withdrawal syndromes. Clo may have a synergistic action with Spm via α2-receptor in the locus coeruleus of the rat brain stem.

Objective: To observe the inhibitory effect of scopolamine(Spm) and chlopromazine (Clo) on withdrawal syndromes in morphine dependent rats. Methods: The intensity of withdrawal syndromes on the model of morphine dependent rats was recorded after single or muiltiple subcutaneous administration(sc) of Spm and Clo at different doses. Results: Withdrawal syndromes were markedly decreased when single Spm 1 mg/kg and Clo 0.5 mg/kg combined with morphine were injected (P<0.05). Spm+Clo(sc) had much stronger effects on inhibiting withdrawal syndromes after intraperitoneal (ip) naloxone in morphine dependent rats (P<0.01). Conclusion: Spm can act on Ach-receptor and relieve morphine withdrawal syndromes. Clo may have a synergistic action with Spm via α2-receptor in the locus coeruleus of the rat brain stem.

Objective To understand the level and distribution of antibody F1 against plague in population of Ningxia natural plague foci in 2007 and 2008. Methods Seven hundred and eighteen blood samples were collected in five major cities and counties of natural plague foci, and 475 blood samples were collected in nonplague area as control group. Conventional indirect hemagglutination, colloidal gold test, and enzyme-linked immunoassay were employed to test the antibody. If the result was tested positive by more than two methods used then the result was defined as positive. Antibody titer that did not reach the positive standard was defined as suspected samples. Results A total of 718 serum samples were tested, the results showed that 9 samples were positive (antibody titer was 1:16 - 1:64), the positive rate was 1.25%(9/718), suspected samples was 28, the detection rate was 3.90%(28/718). Four hundred and seventy-five serum samples in the non-plague area were all negative by the three methods. There was a significant difference of antibody F1 positive rate between residents in historical epidemic area and history nonepidemic area(χ2 = 4.44, P< 0.05). There was no statistical significance of the positive rate[1.25%(9/718), 1.25%(9/718),2.51%(18/718)]among the three methods used(χ2 = 1.91, P> 0.05). Conclusion There still exists a certain proportion of Fl antibody positive people in Ningxia natural plague foci, and these people are distributed in areas where several animal plague prevalent in recent years.

OBJECTIVETo investigate the role of cytoplasmic domain of integrin alpha IIb in platelet signal transduction.

METHODSBinding capacity of integrin alpha IIb(R995A) to antibody platelet activation complex-1 (PAC-1) and pp125 focal adhesion kinase (FAK) phosphorylation of cells were detected by flow cytometry, immune precipitation, and Western blotting.

RESULTSWithout activation, wild-type alpha IIb beta3 Chinese hamster ovary (CHO) cells failed to bind to PAC-1, but mutant chimera alpha IIb(R995A)beta3 CHO cells were able to bind with PAC-1. Furthermore, phosphorylation of pp125 (FAK) in wild-type alpha IIb beta3 CHO cells occured only when cells were adhered to fibrinogen, but could not be detected in bovine serum albumin suspension. However in the mutant chimera group, it could be detected in both conditions.

CONCLUSIONThe mutation in integrin alpha IIb(R995A) alters its affinity state as a receptor, thus also mediating cytoplasmic signal transduction leading to the phosphorylation of pp125 (FAK) without ligand binding.

Animals ; Blood Platelets ; metabolism ; CHO Cells ; Cell Adhesion ; Cricetinae ; Cricetulus ; Cytoplasm ; metabolism ; Dual Specificity Phosphatase 2 ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Humans ; Phosphorylation ; Platelet Glycoprotein GPIIb-IIIa Complex ; genetics ; metabolism ; physiology ; Point Mutation ; Protein Phosphatase 2 ; Protein Tyrosine Phosphatases ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Transfection

BACKGROUNDThe treatment for long bone defects has been a hot topic in the field of regenerative medicine. This study aimed to evaluate the therapeutic effects of calcium sulfate (CS) combined with platelet-rich plasma (PRP) on long bone defect restoration.

METHODSA radial bone defect model was constructed through an osteotomy using New Zealand rabbits. The rabbits were randomly divided into four groups (n = 10 in each group): a CS combined with PRP (CS-PRP) group, a CS group, a PRP group, and a positive (recombinant human bone morphogenetic protein-2) control group. PRP was prepared from autologous blood using a two-step centrifugation process. CS-PRP was obtained by mixing hemihydrate CS with PRP. Radiographs and histologic micrographs were generated. The percentage of bone regenerated bone area in each rabbit was calculated at 10 weeks. One-way analysis of variance was performed in this study.

RESULTSThe radiographs and histologic micrographs showed bone restoration in the CS-PRP and positive control groups, while nonunion was observed in the CS and PRP groups. The percentages of bone regenerated bone area in the CS-PRP (84.60 ± 2.87%) and positive control (52.21 ± 4.53%) groups were significantly greater than those in the CS group (12.34 ± 2.17%) and PRP group (16.52 ± 4.22%) (P < 0.001). In addition, the bone strength of CS-PRP group (43.10 ± 4.10%) was significantly greater than that of the CS group (20.10 ± 3.70%) or PRP group (25.10 ± 2.10%) (P < 0.001).

CONCLUSIONCS-PRP functions as an effective treatment for long bone defects through stimulating bone regeneration and enhancing new bone strength.

Animals ; Bone Regeneration ; drug effects ; Calcium Sulfate ; pharmacology ; Male ; Platelet-Rich Plasma ; Rabbits

OBJECTIVETo evaluate the relationship between lifestyle habits and the components of metabolic syndrome (MS).

METHODSBased on the routine health check-up system in a certain Center for Health Management of Shandong Province, a longitudinal surveillance health check-up cohort from 2005 to 2010 was set up. There were 13 225 urban workers in Jinan included in the analysis. The content of the survey included demographic information, medical history, lifestyle habits, body mass index (BMI) and the level of blood pressure, fasting blood-glucose, and blood lipid, etc. The distribution of BMI, blood pressure, fasting blood-glucose, blood lipid and lifestyle habits between MS patients and non-MS population was compared, latent variables were extracted by exploratory factor analysis to determine the structure model, and then a partial least squares path model was constructed between lifestyle habits and the components of MS.

RESULTSParticipants'age was (46.62 ± 12.16) years old. The overall prevalence of the MS was 22.43% (2967/13 225), 26.49% (2535/9570) in males and 11.82% (432/3655) in females. The prevalence of the MS was statistically different between males and females (χ(2) = 327.08, P < 0.01). Between MS patients and non-MS population, the difference of dietary habits was statistically significant (χ(2) = 166.31, P < 0.01) in MS patients, the rate of vegetarian, mixed and animal food was 23.39% (694/2967), 42.50% (1261/2967) and 34.11% (1012/2967) respectively, while in non-MS population was 30.80% (3159/10 258), 46.37% (4757/10 258), 22.83% (2342/10 258) respectively. Their alcohol consumption has statistical difference (χ(2) = 374.22, P < 0.01) in MS patients, the rate of never or past, occasional and regular drinking was 27.37% (812/2967), 24.71% (733/2967), 47.93% (1422/2967) respectively, and in non-MS population was 39.60% (4062/10 258), 31.36% (3217/10 258), 29.04% (2979/10 258) respectively. The difference of their smoking status was statistically significant (χ(2) = 115.86, P < 0.01) in MS patients, the rate of never or past, occasional and regular smoking was 59.72% (1772/2967), 6.24% (185/2967), 34.04% (1010/2967) respectively, while in non-MS population was 70.03% (7184/10 258), 5.35% (549/10 258), 24.61% (2525/10 258) respectively. Both lifestyle habits and the components of MS were attributable to only one latent variable. After adjustment for age and gender, the path coefficient between the latent component of lifestyle habits and the latent component of MS was 0.22 with statistical significance (t = 6.46, P < 0.01) through bootstrap test. Reliability and validity of the model:the lifestyle latent variable: average variance extracted was 0.53, composite reliability was 0.77 and Cronbach's a was 0.57. The MS latent variable: average variance extracted was 0.45, composite reliability was 0.76 and Cronbach's a was 0.59.

CONCLUSIONUnhealthy lifestyle habits are closely related to MS. Meat diet, excessive drinking and smoking are risk factors for MS.

Adult ; Female ; Humans ; Life Style ; Longitudinal Studies ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Models, Statistical ; Prevalence ; Risk Factors

A new method for establishing ES cell lines from 129/ter. C57BL/6J mice was set up which was characterized by the murine embryonic fibroblast cell(MEF) feeder, the medium of rat heart cell-conditioned medium(RH-CM) for ES cells, and the consecutive digestion by the digestion liquid containing 1% serum. Every group of improved experiments was done with a control of routine method. The results showed that, compared with routine method, the improved way increased the ratio of ES cell lines of 129/ter mice from 11.8% to 33.3%, and of C57BL/6J from 3.7% to 13.3%. The difference is distinct. The passage culture of ES cells showed that, compared with medium added LIF, RH-CM not only inhibited the differentiation of murine ES cells, maintained its dipoild karyotype, but also promote its adherence growth. This kind of culture condition not only maintained the ES cells in an undifferentiated state and their normal dipoild karyotype, but also a series of other characteristics of totipotent embryonic stem cells during extended culture period.
Alkaline Phosphatase ; metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Cell Differentiation ; Cell Division ; Cell Line ; Embryo, Mammalian ; cytology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Rats ; Stem Cells ; physiology