In this study, the human umbilical vein endothelial cell model was used to study the regulating effect of lipophilic components in Salvia miltiorrhiza on angiogenesis, and explore its possible mechanism. The cell model was established to determine the effect of lipophilic components in S. miltiorrhiza on the proliferative activity and migration capacity of endothelial cells. Then the realtime fluorescence quantification PCR technology was applied to detect the changes in the gene expressions of angiogenesis-related cytokines VEGF-A, VEGF-C and MMP-9. The results showed that 5 mg x L(-1) lipophilic components in S. miltiorrhiza could inhibit the proliferation and migration of endothelial cells, and reduce the expression of VEGF-A and MMP-9 genes. It indicated that lipophilic components in S. miltiorrhiza may inhibit the proliferation and migration of endothelial cells by inhibiting the expression of VEGF-A and MMP-9 genes, so as to show the inhibitory effect on angiogenesis.
Angiogenesis Inhibitors ; chemistry ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

In this study, five rhesus macaques were inoculated intravenously with SIVmac251 to establish a model of simian autoimmune deficiency syndrome (SAIDS). Peripheral blood samples were collected at different time points to monitor changes in the total T cell number and T lymphocyte subset. Plasma viral loads, cytokine expression levels and anti-SIV antibody levels were also assayed to acquire certain basic indexes to evaluate disease progression in the rhesus macaque SAIDS model. During the acute stage of infection, plasma viral loads reached a peak at week 1 post-inoculation and lasted for approximately 3 to 44 weeks. The CD3+ CD4+ T lymphocyte count in peripheral blood also transitorily decreased. During the same period, the level of interferon-gamma show an increasing trend, whereas IL-12 levels decreased; IL-2, IL-4, IL-10 and TNF-alpha were maintained at normal levels or could not be detected. During the asymptomatic and ARC phases, plasma viral loads persisted above 10(4) RNA copies/mL and either increased or declined during the later stages of disease; CD3+ CD4+ counts showed a steadily declining trend and the ratio of CD4 to CD8 decreased during late-stage disease. Moreover, antibodies against viral proteins were detected in the plasma and showed a significant increasing trend, while there were no apparently changes in the levels of IFN-gamma, IL-12, IL-2, IL-4, IL-10 and TNF-alpha. In conclusion, the characteristics of the SIV animal models in our study are similar to those of patients with AIDS. Therefore, the rhesus macaque SIVmac251 infection models can be applied for further studies into AIDS.
Animals ; Antibodies, Viral ; blood ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; virology ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; HIV Infections ; genetics ; immunology ; virology ; HIV-1 ; physiology ; Humans ; Macaca mulatta ; Male ; Simian Acquired Immunodeficiency Syndrome ; genetics ; immunology ; virology ; Simian Immunodeficiency Virus ; physiology ; Viral Load

OBJECTIVETo investigate the effect of adenosine A2A receptor knockout (A(2A)RKO) on relationship between continuous activation of phospho-c-Jun N-terminal kinase (P-JNK) and expression of nerve cell apoptosis in hippocampus CA1 domain of newborn mice after hypoxia/ischemia brain damage(HIBD) and its potential mechanism.

METHODSA(2A)RKO mice and adenosine A2A receptor wildtype (A(2A)RWT) littermates (n = 80) were divided into Sham operation group (S) and model group (M), 1, 3 and 7 day after HIBD, totally 8 groups. HIBD was developed with 7 day-old neonatal mice according classical Rice-Vannucci method. It was tested the effect of A(2A)RKO on short-term neurofunctional outcomes consisted of three developmental reflexes (righting, geotaxis and cliff aversion), the changes of brain pathology with hematoxylin-eosin (HE) staining and Nissl staining, the expressions of nerve cell apoptosis with terminal deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling(TUNEL) staining and P-JNK were observed by immunohistochemistry.

RESULTSThe neurological behavior injuries and brain histopathological damages and nerve apoptosis cells were aggravated in A(2A)RKO newborn mice after HIBD. The positive expressions of P-JNK were significantly higher in the ischemic hippocampus CA1 domain after HIBD than ones in group S respectively (P < 0.01), reaching to peak at 1 day and then began gradually decreasing. P-JNK expression in model knockout(MKO) at 1, 3 and 7 day increased greatly compared to those in the previous time point of corresponding model wildtype (MWT) (P < 0.01, P < 0.05, P > 0.05); there was a positive correlation between the expressions of P-JNK and nerve cell apoptosis after HIBD in newborn mice(r = 0.837, P < 0.01).

CONCLUSIONEarly continuous activation of P-JNK might be involved in the aggravated nerve apoptosis cells and brain damage induced by A(2A) RKO newborn mice after HIBD.

Animals ; Animals, Newborn ; Apoptosis ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Knockout ; Neurons ; drug effects ; metabolism ; pathology ; Receptor, Adenosine A2A ; genetics

Female ; Humans ; Liver Failure, Acute ; blood ; Liver Neoplasms ; blood ; Liver Transplantation ; Male ; Thrombelastography ; Whole Blood Coagulation Time

Rhesus monkeys with high specific H5N1 antibody were inoculated the second time with H5N1 virus, the result of the second time H5N1 inoculation and the effect of first time H5N1 inoculation on second inoculation was evaluated. Monkeys of NO. 3, NO. 4, NO. 5 were inoculated with H5N1 allantoic fluid and NO. 6 with noninfectious allantoic fluid by intratracheal thyrocricoid puncture. Three months later, NO. 4, NO. 5, NO. 6 monkeys were infected with 7 ml TCID50 10(4.875) H5N1 allantoic fluid and NO. 3 monkey with 7 ml noninfectious allantoic fluid at the same time by the same method. Clinical symptoms were recorded and antibody response was detected by ELISA. NO. 3, NO. 4, NO. 6 monkeys were killed after 72 h post infection and NO. 5 monkey was killed after 7 days post infection. Pathologic changes of the infected monkeys' lung were examined by HE staining,immunohistochemistry and the virus in lung was detected by RT-PCR. Results showed that NO. 3, NO. 4, NO. 5 monkeys still retained high level of specific antibody, H5N1 virus only could be detected in NO. 6 monkey's lung by immunohistochemistry and RT-PCR ,and the lung of NO. 6 monkey injured worst . It can be concluded that Rhesus monkeys inoculated with H5N1 avian influenza A virus at the first time could retain a high level of specific antibody in 90 days and the clinical symptom had almost recovered, the ability of Rhesus monkeys to resist second infection of H5N1 virus was enhanced notably at that moment.
Animals ; Antibodies, Viral ; blood ; immunology ; Enzyme-Linked Immunosorbent Assay ; Immunohistochemistry ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; pathogenicity ; Macaca mulatta ; Monkey Diseases ; immunology ; pathology ; virology ; Orthomyxoviridae Infections ; blood ; immunology ; pathology ; Reverse Transcriptase Polymerase Chain Reaction

Objective: This study was designed to investigate the prognostic value of serum β2-micorglobulin(β2-MG) level in non-Hodgkin s lymphoma(NHL) patients and its correlation with Ann Arber staging system and the pathological type. Method: Serum β2-MG levels in 75 de novo NHL patients were measured by radioimmunoassay. Results: The β2-MG level in 75 NHL patients was higher than that in nomal control level in 52％ of the patients and correlated with the Ann Arber stage. There was significant difference between two groups (P<0.01). The international prognostic index (IPI) and the response rate to chemotherapy were also correlated with the β2-MG level in serum. The response rate, the complete response, and partial response were 87.2%, 61.5%, and 25.6% in normal serum β2-MG group, while in the high β2-MG group they were only 75.0%, 50.0%, and 25.0%, respectively, which were significantly different between the two groups (P<0.05). The difference of 5-year survival between the two groups was significant (64.1% vs 27.8%, P<0.05) and favorable in normal β2-MG group. Conclusion: β2-MG level in serum is an independen prognostic factor in de novo NHL, and the patients with abnormal β2-MG level in serum have a unfavorable prognosis.

Objective To evaluate the clinical effects of the bone-ring augmentation in implant restoration using the bone ring technique.Methods Sixteen patients (30 teeth) with severe alveolar bone absorption were selected.Bone rings (circular osteotomies) were outlined at the symphysis area and a central osteotomy for implant placement was done using the bone ring drills before its removal.The rings were then removed and sculptured to fit the alveolar ridge and then covered by Bio-oss and Bio-Gide membrane.This was followed by screwing the implant through the ring,gaining its primary stability from the prepared basal bone.After 4-6 months,the implant was exposed and abutment connected for temporary restoration.Results Of the 16 patients,14 cases were healed by first intention,2 cases were infected after the bone ring implantation resulting in some bone absorption but healed after bone grafting materials were placed at the second stage of the onlay procedure.The bone ring implantation showed no absorption and no implant failure.After the bone grafting the alveolar bone height increased by 4.1 mm on average and the thickness increased by 3.27 mm on average.Conclusions Implant bone ring can effectively augment the absorbed alveolar ridge.The procedure has the advantages of bone removal,grafting and implantation at one stage.

OBJECTIVEChildren aged 3 - 6 years in the urban areas of China were surveyed for the first time to find out the state of child neglect (CN) as well as the major relevant risk factors so as to provide evidence for developing intervention measures.

METHODS1163 children (of whom 49.6% were males and 4.5% were minority nationality) were randomly sampled under multistage stratification, from 25 cities which representing 15 provinces of China. Based on the Child Neglect Norms used by China, prevalence of CN was identified and SPSS-Windows 11.0 was employed for statistical analysis. Scores, frequency/degrees, age, sex and 5 types (physical, emotional, educational, medical and safety) of CN on every group of the regions, were calculated. Multifactorial analysis was conducted through Binary Logistic Regression and multiple linear regression to determine the relevant risk factors.

RESULTS(1) The average degree of CN for the 3 - 6 year-olds was 42.2, with its prevalence as 28.0%. Degrees of CN for the groups of 3, 4, 5, 6-year-olds were 41.7, 42.2, 42.1 and 43.1 (F = 0.988, P > 0.05), with frequencies of 25.0%, 25.3%, 27.9% and 35.4% (chi(2) = 4.798, P > 0.05), respectively. Degrees for CN in males and females were 42.7 and 41.8 (F = 2.502, P > 0.05) with the frequencies as 32.6% and 23.7% (chi(2) = 6.585, P < 0.05), respectively. Degrees of CN for the five types were 39.4-43.4 with the frequencies as 5.1%-12.9%, respectively. No significant difference was found in the frequency of the types (with an exception on 'physical neglect') between males and females (P > 0.05). The highest frequency (42.9%) of CN was seen in the single-parent families and the lowest in large family with three generations (25.5%). (2) According to monofactorial chi(2) test, the possible risk factors of CN would include: educational background, occupation and decrease of income of the parents during last year, etc. (3) Binary Logistic regression analysis showed that the influential factors to the occurrence of CN would include: father's educational background, sex of the child and mother's occupation, etc. (4) Multiple linear regression showed that the influential factors to the degree of CN were: family structure, number of supporting family members, relationship between parents and children, etc.

CONCLUSIONThe degree and frequency of CN among children aged 3 to 6 in the urban areas of China were high but similar among the four age groups. Male children had a higher frequency of neglect than females, but with similar degree. Children in single-parent families had the highest frequency. The major influential factors of CN would include: educational background, occupation, family structure, family income of the parents which were similar to the results reported from foreign literature.

Child ; Child Abuse ; statistics & numerical data ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Parenting ; Parents ; psychology ; Social Class ; Surveys and Questionnaires ; Urban Population

OBJECTIVETo develop a novel rapid protocol for the detection of occult disseminated tumor cells by a recombinant herpes simplex virus expressing GFP (HSV(GFP)).

METHODSTumor cells of seven cell lines were exposed to HSV(GFP) and then examined for GFP expression by fluorescence microscopy. Various numbers of tumor cells (10, 100, 1000, 10 000) were mixed into 2 ml human whole blood, separated with lymphocytes separation medium, exposed to HSV(GFP), incubated at 37°C for 6 - 24 h and then counted for the number of green cells under the fluorescence microscope. Some clinical samples including peripheral blood, pleural effusion, ascites, spinal fluid from tumor-bearing patients were screened using this protocol in parallel with routine cytological examination.

RESULTSHSV(GFP) was able to infect all 7 tumor cell lines indicating that the HSV(GFP) can be used to detect different types of tumor cells. The detection sensitivity was 10 cancer cells in 2 ml whole blood. In the clinical samples, there were 4/15 positive by routine cytological examination but 11/15 positive by HSV(GFP), indicating a higher sensitivity of this new protocol.

CONCLUSIONRecombinant herpes simplex virus-mediated green fluorescence is a simple and sensitive technique for the identification of occult disseminated cancer cells including circulating tumor cells (CTCs).

Animals ; Cell Line, Tumor ; Cercopithecus aethiops ; Green Fluorescent Proteins ; metabolism ; Humans ; Neoplastic Cells, Circulating ; metabolism ; pathology ; Recombinant Proteins ; metabolism ; Sensitivity and Specificity ; Simplexvirus ; metabolism ; Vero Cells

OBJECTIVENucleic acid amplification (PCR) fluorogenic quantitative assay is used for the diagnosis of respiratory syncytial virus (RSV) infection. This study was designed to explore the sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection and RSV infection conditions by detecting the presence of RSV-RNA related sequences in children.

METHODSBronchial and nasopharyngeal secretions specimens from 261 hospitalized children with respiratory tract infections from January 2007 to October 2008 were collected. Respiratory syncytial virus nucleic acid (RNA) in the specimens was measuredby PCR fluorogenic quantitative assay. Blood RSV-IgM was detected by enzyme linked immunosorbent assay (ELISA). The sensitivity for ascertaining respiratory RSV infection was compared between the two assays.

RESULTSThe RSV-RNA positive rate ascertained by PCR fluorogenic quantitative assay (38.7%) was significantly higher than blood RSV-IgM positive rate (21.1%) (p<0.01). The RSV-RNA positive rate (43.6%) in children at ages of less than 6 months was significantly higher than that in children at ages of 1 to three years (32.1%) (p<0.01). The RSV-RNA positive rate in children with bronchiolitis (58.5%) was the highest, followed by bronchopneumonia (38.2%) and acute bronchitis (20.0%).

CONCLUSIONSThe sensitivity of PCR fluorogenic quantitative assay for ascertaining respiratory RSV infection is higher. RSV is a major pathogen of lower respiratory tract infections in infants and young children. A higher rate of RSV infection is associated with a younger age. RSV infection is the most common in children with bronchiolitis.

Antibodies, Viral ; blood ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescence ; Humans ; Immunoglobulin M ; blood ; Infant ; Male ; Polymerase Chain Reaction ; methods ; RNA, Viral ; analysis ; Respiratory Syncytial Viruses ; genetics ; immunology ; isolation & purification ; Respiratory Tract Infections ; virology ; Sensitivity and Specificity ; Sputum ; virology