Herpes simplex virusⅠ(HSVⅠ) regulating the pathway of transcription and translation modify in host cell is a very systematic and complicate system. A clear understanding of the concrete mechanisms of infection will greatly help to comprehend the virus replication and the interaction with the host cell. By the analysis of 2-DE, the heterogeneous nuclear ribonucleoprotein H2 in human fetal liver cell represent distinction after the HSVⅠinfection.Utilization of Northern blot and Western blot technologies verified the expression of hnRNP H2 in different stage of virus infection is varied.

OBJECTIVETo construct a recombined phage vaccine and to evaluate the efficiency of this phage vaccine against EGFR-positive tumors.

METHODST7 phage display system was used to display five fragments of the extracellular domain of chicken EGFR. The EGFR was expressed as a fused protein on the surface of the T7 phage 10B capsid protein. The EGFR expression of the phage vaccine was verified by Western-blot analysis. Anti-EGFR antibody was detected by ELISA. Splenic lymphocytes of the immunized mice were separated and used to determine the immunotoxic effect against A431 cells. The phage vaccines were injected into C57 mice 4 times before Lewis lung cancer cells inoculation. Tumor volume was recorded to evaluate the anti-tumor effect of each vaccine.

RESULTSFive phage vaccines inserted with the chicken EGFR gene were successfully constructed. Western blot assay showed that the extracellular domain of chicken EGFR proteins were displayed on the surface of the phage. Specific antibody was induced in the immunized mice, compared with the control group. Splenic lymphocytes of the immunized mice were shown to be immunotoxic against A431 cells. The killing rates of the experimental groups were higher than that of control group (P < 0.001, t-Student test). The highest killing rate was (45.74 +/- 7.21)%. The tumor growth was inhibited in the experimental groups compared with those of control groups (P < 0.05 in C1, C2, C3, C4 groups, P > 0.05 in C5 group).

CONCLUSIONOur results demonstrated that recombined EGFR phage vaccines may be used to induce therapeutic anti-tumor immunity against EGFR-positive tumors.

Animals ; Bacteriophage T7 ; genetics ; Blotting, Western ; Cancer Vaccines ; administration & dosage ; genetics ; immunology ; Capsid Proteins ; genetics ; metabolism ; Carcinoma, Lewis Lung ; immunology ; pathology ; therapy ; Cell Line, Tumor ; Chickens ; Cytotoxicity Tests, Immunologic ; Immunotherapy ; methods ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Random Allocation ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

Objective: To explore the relationship between plasma Jagged1 protein level and coronary collateral circulation (CCC) formation in patients with coronary artery disease(CAD). Methods: According to coronary angiography (CAG) examination, our research was categorized in 2 groups: CAD group, n=89 patients with at least one of left anterior descending (LAD), left circumflex(LCX) or right coronary artery(RCA) stenosis ≥ 95％ and Control group, n=30 subjects without abnormal findings by CAG. Based on Rentrop grading system, CAD group was further divided into 2 subgroups: Good CCC subgroup, n=42 patients with Rentrop grade ≥ 2 and Poor CCC subgroup, n=47 patients with Rentrop grade≤1. Plasma levels of Jagged1 protein,vascular endothelial growth factor (VEGF) were measured by ELISA and the relevant correlation study was conducted by multivariate regression analysis. Results: Compared with Control group, CAD group had increased plasma levels of Jagged1 protein (38.74±10.60)ng/L vs (23.04±8.97)ng/L and elevated VEGF (113.98±30.80)pg/L vs (72.73±14.55)pg/L. Compared with Poor CCC subgroup, Good CCC subgroup presented increased Jagged1 protein (46.77±8.49)ng/L vs (31.56±6.26)ng/L and elevated VEGF (128.10±20.24) pg/L vs (92.43±21.09)pg/L. Correlation study showed that Jagged1 protein was positively related to VEGF in CAD patients (r=0.730, P<0.01); multivariate regression analysis indicated that Jagged1 protein (OR=1.318, P=0.000) and VEGF (OR=1.043, P=0.043) were the independent predictors for CCC processing.Conclusion: CAD patients with good CCC had the higher plasma Jagged1 protein level than the patients with poor CCC which implied that Jagged1 protein played important role in CCC processing, such finding may provide a new direction for treating CAD patients in clinical practice.

OBJECTIVETo appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation.

METHODSTwenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment.

RESULTSAfter separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05).

CONCLUSIONThe single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.

Cell Separation ; methods ; Centrifugation, Density Gradient ; methods ; Humans ; Male ; Povidone ; Silicon Dioxide ; Sperm Count ; methods ; Spermatozoa ; cytology

OBJECTIVETo compare the effect of different antegrade pulmonary blood flow on the further development of pulmonary artery after Glenn procedure in cyanotic congenital heart defects (CHD) patients.

METHODSBetween October 2000 and December 2006, 132 CHD patients with decreased pulmonary artery blood flow underwent bidirectional Glenn shunt, among them 18 patients received intraoperative lung biopsy. Patients were divided into two groups according to their different sources of antegrade pulmonary blood flow: antegrade arterial blood flow group (n = 33) and antegrade venous blood flow group (n = 99). The percutaneous oxygen saturation (SpO2), hemoglobin (Hb) concentration, and hemotocrit (Hct) value were examined and recorded before and after operation. The diameters of left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured with two-dimensional echocardiography and the results were used to calculate the pulmonary artery index (PAI). The method of half-quantitative morphometric technique and an image analyzer were used to measure the following indicators of pulmonary microvessels: the percentage of media thickness (MT%), the percentage of media section area (MS%), vascular numbers of per square centimeter (VPSC), and mean alveolar number (MAN).

RESULTSBefore the operation, obvious cyanosis was found in both groups, while SpO2, Hct, and Hb were not significantly different (P > 0.05). LPA, RPA, and PAI were not significantly different between two groups (P > 0.05). The MT% and MS% in antegrade venous blood flow group were significantly less than those in antegrade arterial blood flow group (P < 0.05), but VPSC and MAN were not significantly different (P > 0.05). After Glenn procedure, hypoxia and cyanosis were remarkably improved in both two groups. There was a significantly negative correlation between SpO2 and Hct (r = -0.49, P < 0.01) or Hb (r = -0.196, P < 0.01 ). The PAI increased by 22% in antegrade arterial blood flow group and 44% in antegrade venous blood flow group (P < 0.05). The diameters of LPA and RPA in antegrade venous blood flow group were significantly larger than those in antegrade arterial blood flow group (P < 0.05) and the growth of RPA in antegrade arterial blood flow group was not significant.

CONCLUSIONA better pulmonary artery growth occurs in the patients of pulmonary stenosis after Glenn shunt than in those of pulmonary atresia, and it contributes to an earlier completion of Fontan procedure.

Blood Flow Velocity ; Cardiac Surgical Procedures ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; surgery ; Humans ; Infant ; Male ; Pulmonary Artery ; growth & development ; physiopathology ; surgery ; Pulmonary Veins ; physiopathology ; Treatment Outcome

OBJECTIVETo determine the contents of NOS and endogenous CO in the penile corpus cavernosum of Wistar rat models of hyperthyroid and hypothyroid, to detect the effects of thyroid hormone and endogenous CO on rat penile erection, and to further investigate the influence of thyroid hormone on human penile erection.

METHODSFifty 3-month-old male Wistar rats were randomly and equally divided into 5 groups: a hyperthyroid, a hyperthyroid treatment, a hypothyroid, a hypothyroid treatment and a control group. The contents of NOS and CO were detected by ultraviolet spectrophotometry.

RESULTSCompared with the controls, the content of NOS was significantly decreased in both the hyperthyroid and the hypothyroid groups (P < 0.01), even lower in the latter than in the former (P < 0.01), and so was the content of CO (P < 0.01), lower in the former than in the latter (P < 0.01). After treatment, the levels of NOS and CO were both elevated in either the hyperthyroid or the hypothyroid group, with no significant difference from that of the controls (P > 0.05).

CONCLUSIONThe contents of NOS and CO in the penile corpus cavernosum decrease with the imbalance of thyroid hormones, but return to normal once the imbalance is corrected. Under the same conditions, hypothyroid inflicts more damage on penile erection than hyperthyroid.

Animals ; Carbon Monoxide ; metabolism ; Disease Models, Animal ; Hyperthyroidism ; drug therapy ; Hypothyroidism ; drug therapy ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Thyroid Hormones ; pharmacology ; therapeutic use

OBJECTIVETo study the association of the changes of serum insulin, insulin-like growth factor (IGF-1), insulin-like growth factor binding proteins(IGFBPs), body mass index (BMI), waist and hip circumference ratio(WHR) with the genesis of colorectal cancer.

METHODSSera from 244 colorectal cancer patients before operation, 371 patients after operation and 150 healthy subjects were assayed for insulin, leptin, IGF-1, IGFBP-1 and IGFBP-3 by the enzyme-linked immunosorbent assay. SPSS 13.0 statistics software was applied to analyze the data.

RESULTSThe serum levels of insulin, IGF-1 and the ratio of IGF-1/ IGFBP-3 in colorectal cancer patients before and after surgical treatment were significantly higher than those in controls. The serum levels of IGFBP-3 in patients before and after operation were significantly lower than those in controls, and the differences were significant(P=0.015,P=0.001, respectively). The BMI in colorectal carcinoma patients was not significantly different to the healthy controls(P>0.05). The WHR in colorectal carcinoma patients was higher than that in healthy subjects, and the difference was significant(P=0.003, P=0.035 respectively). The WHR in colon cancer patients was different to that in rectal cancer patients(P=0.046). The WHR and BMI in colon carcinoma patients were positively correlated with the serum insulin level and the value of IGF/IGFBP3. The WHR and BMI were negatively correlated with IGFBP3. The WHR and BMI were not correlated with IGF-1 and IGFBP1.

CONCLUSIONSThe serum insulin, IGF-1 levels and the value of IGF-1/IGFBP-3 are significantly increased in colorectal cancer patients, and serum IGFBP-3 level is markedly decreased, which may be related to the genesis of colorectal cancer, but are not correlated with the progress and improvement of colorectal cancer. Central adipositas may be a risk factor for the genesis of colon cancer.

Adult ; Aged ; Body Mass Index ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; Female ; Humans ; Insulin ; blood ; Insulin-Like Growth Factor Binding Protein 1 ; blood ; Insulin-Like Growth Factor Binding Protein 3 ; blood ; Insulin-Like Growth Factor I ; metabolism ; Leptin ; blood ; Male ; Middle Aged ; Risk Factors ; Waist-Hip Ratio

Objective To report a large family with an autosomal dominant dementia associated with mutation in the prion protein gene(PRNP)and the detailed clinical,neuroimaging and pathological manifestations.Methods Two patients from a large family of dementia were admitted to our ward and the data of their medical history,physical examination,video electroenceplialogram,neuroimaging were colleted.A sterotactic biopsy of the right frontal lobe of the proband was done.After the informed consent from the family members obtained,the genomic DNA was extracted from peripheral blood leucocytes of 5 persons followed by in,vitro amplification using polymerase chain reaction(PCR).The PCR products were directly sequenced by Sanger method.PRNP gene sequence was also examined in 150 normal Chinese to exclude single nueleotide polymorphism.Results A missense mutation of PRNP gene in 5 farnily members was detected,resulting in Gll4V mutation in the prion protein,with M/M genotype of eodon 129.This mutation was not detected in 150 normal Chinese.The proband was diagnosed as inherited prion disease by her clinical features,including neuropsychiatrie disturbances and progressive dementia,and manifestations of neuroimaging,EEG,neuropathology and PRNP gene mutation.Conclusion The first autosomal dominant pedigree of family prion disease is found in China with G114V mutation in PRNP gene which may lead to the prion disease directly.

As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since the ICP0 encoding sequence contains many rare codons which are absent in E.coli, and ICP0 is highly unstable in prokaryotic cells, expression of entire ICP0 in prokaryotic cells has never been reported. In order to further investigate the function of ICP0, a recombinant plasmid was constructed by subcloning a cDNA fragment encoding an amino-terminal of 105 residues of the ICP0 protein into pGEX-5x-1 vector. The resulting GST-105 fusion antigen peptide was expressed with high efficiency in E.coli. Antibodies prepared after the immunization of mice with purified fusion protein can recognize not only the denatured ICP0 protein, but also the native ICP0 protein with normal biological conformation.

Objective To explore the protective effects of diazoxide on injury of human renal tubular cell(HK-2)induced by serum obtained from neonates with asphyxia.Methods HK-2 cells was used as the target cel1.The attacking concentration of serum obtained from neonates with asphyxia was 200 mL/L.The experiment was designed as 3 groups.HK-2 cells were divided into control group,asphyxia group,and diazoxide group.Control group:joined nutrient fluid including 100 mL/L embryo cow blood serum.Asphyxia group:joined nutrient fluid including the isometric 200 mL/L serum obtained from neonates with asphyxia.Diazoxide group:the diazoxide was joined nutrient including the isometric 200 mL/L serum obtained from neonates with asphyxia fluid.The diazoxide density finally was 100 ?mol/L.Then the change of morphology was observed and photographed under inverted microscope,and the cell viability was measured by methyl thiazolyl tetrazolium method,and the leakage rate oflactate dehydrogenase(LDH)was determined by biochemical methods.Results Under inverted microscopy,HK-2 cells in control group pastes the wall to be good,assumes the paving stone type,into flat polygon,fission many,the cell arrangement was close,connection large expanse,quantity were many.Compared with control group,the HK-2 cell to suffer injury obviously,the shape changed,become the anomalous circular or the ellipse by the model flat polygonal cell,the intercellular space crevice enlarged,the connection was loose,intercellular space obviously many cell fragmented.Living cell quantity reduced obviously,the cell vigor dropped,and the leakage rate of LDH increased significantly in asphyxia group(P