OBJECTIVE: Using the method of lactate dehydrogenase (LDH) assay, to observe the activities of rat cerebral microvascular endothelial cells (CMECs) intervened by Tongluo Jiunao Injection (TLJNI), a traditional Chinese compound drug removing toxin to dredge brain collaterals, and then further study the effects of different kinds of conditioned mediums (CMECs-CM) of cerebral microvascular endothelial cells on ischemia and ischemia/reperfusion cerebral cortex cells, and to probe into the drug pharmacological mechanisms of CMECs in modulating the neurons. METHODS: Three kinds of CMECs (normal, ischemic and ischemic/reperfusional) were all treated by TLJNI previously, and then the three pairs of CMECs-CM without serum were collected respectively for LDH assay. Rat cerebral cortex neurons were also primarily cultured and then divided into similar three groups (normal, ischemic and ischemic/reperfusional). The neuron responses caused by CMECs-CM at different concentrations were observed by using LDH transudation rate assay. RESULTS: The LDH release values of ischemic and ischemic/reperfusional CMECs with TLJNI treatment were obviously reduced (P<0.01) compared with the same kinds of CMECs untreated. For ischemic neurons, both conditioned medium of ischemic CMECs (Is-CM) and conditioned medium of ischemic CMECs with drug treatment (IsT-CM) in high concentration of 100% increased the LDH transudation rate (P<0.01), while in low concentration of 10%, IsT-CM reduced the transudation rate (P<0.05). For ischemia/reperfusion neurons, all kinds of CMECs-CM reduced the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, 10% or 50% showed relatively stronger effects, and both conditioned medium of normal CMECs (N-CM) group and conditioned medium of ischemic/reperfusional CMECs (Rp-CM) group had statistical significance (P<0.05 or P<0.01). For normal neurons, all kinds of CMECs-CM increased the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, only conditioned medium of normal CMECs (N-CM) had statistical significance (P<0.05 or P<0.01). CONCLUSION: The study shows that TLJNI is capable of preventing the damage of CMECs from both ischemia and ischemia/reperfusion states. Chinese drug can restrain the brain ischemia and ischemia/reperfusion damage by the media that CMECs modulate the neurons, demonstrating the pharmacological mechanisms of TLJNI. This work also indicates that there exist some active substances against ischemia/reperfusion injury secreted from CMECs-CM with TLJNI treatment.

Objective To elucidate the interaction between osteoblast of bone marrow microenvironment and leukemia cells,and to investigate the role of osteoblast in the leukemia cells survival and apoptosis and the influence of leukemia cells on the osteoblast.Methods Leukemia cells from AML1-ETO9a-Rac1 mouse leukemia model and osteoblast cells were used.The ratio of GFP+ leukemia cells that co-cultured with or without osteoblast was detected by FACS.In addition,the apoptosis level of leukemia cells was detected by flow cytometry by PI and Annexin Ⅴ labeling.Activation level of PARP was determined by Western-blot.Real-time PCR (RT-PCR) was utilized to detect the mRNA level of TPO,N-cadherin,OPN and Ang1 in osteoblast which was separated from leukemic mice.Results The ratio of GFP+ cells in AE9a-Rac1 leukemia cells co-cultured with osteoblast cell was significantly higher than that of AE9a-Rac1 leukemia cells cultured alone.The apoptotic level of AE9a-Rac 1 leukemia cells cultured alone was significant higher than that of AE9a-Rac 1 leukemia cells in co-culture system.Western blot showed that activated level of PARP in AE9a-Rac1 leukemia cells co-cultured with osteoblast was lower than that cultured alone.RT-PCR result showed that TPO and N-cadherin mRNA levels in primary osteoblast separated from leukemic mice were higher than that from normal mice.Ang1 and OPN mRNA levels of osteoblast from leukemia mice were lower.Conclusion Osteoblast cell can support the survival and inhibit the apoptosis of leukemia cells.Leukemia cells can influence the functions of osteoblast by microenvironment associated cytokines production.

Objective To explore the effect of comprehensive intervention technique in neuroses nursing.Methods 48 inpatient neuroses were randomly divided into observation group(drug treatment and comprehensive intervention technique)and control group(drug treatment),using self-rating anxiety scale(SAS)and self-rating depression scale(SDS)to evaluate clinical symptoms before and after 2 months treatment.Results Before treatment.SAS and SDS scores between two patient groups had no statistic difference(P>0.05),but after 2 months treatment.the observation group had lower SAS and SDS scores than the control group,the difference were obvious(P<0.05).Conclusions Comprehensive intervention technique was useful in neuroses nursing which can ameliorate depression and anxiety emotions,improve illness recover.

Chloroplast genome sequences have comprehensive application prospects in DNA barcoding and chloroplast engineering in traditional Chinese medicine. The complete chloroplast genome of Magnolia officinalis sequenced by high-throughput pyrosequencing and a sequencing procedure was established. Fourteen contigs were obtained after de nove assembly. The sequencing percent of coverage was 99.99%. The chloroplast genome is 160 183 bp in size, and has a typical quadripartite structure with the large (LSC, 88 210 bp) and small copy (SSC, 18 843 bp) regions separated by two copies of an inverted repeat (IRs, 26 565 bp each). chloroplast genes were successfully annotated, of which 17 genes located in each IR region. The chloroplast genome features in Magnolia officinalis are nearly identical to those from other Magnoliid chloroplast genomes. Phylogenetic analyses were performed based on 81 shared coding-genes for a total of 9 Magnolia samples of 5 closely related species. Results showed that distinguishing among species was generally straightforward at the species and population level. This study confirmed the effectiveness of our chloroplast genome sequencing procedure. The chloroplast genome can provide distinguishing differences to help identify Magnolia officinalis and its closely related plants.
Base Sequence ; Chloroplasts ; genetics ; DNA, Chloroplast ; genetics ; Genes, Chloroplast ; Genes, Plant ; Genome, Chloroplast ; Genome, Plant ; High-Throughput Nucleotide Sequencing ; Magnolia ; classification ; genetics ; Phylogeny ; Sequence Analysis, DNA

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

Administration, Sublingual ; Bacterial Vaccines ; administration & dosage ; immunology ; Child ; Child, Preschool ; Double-Blind Method ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin A, Secretory ; analysis ; Male ; Pseudomonas Vaccines ; Recurrence ; Respiratory Tract Infections ; immunology ; prevention & control ; Treatment Outcome

Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Arteriovenous Malformations ; complications ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Multiple Sclerosis ; Spinal Cord Diseases ; complications ; metabolism ; pathology

Objective To understand the status and influential factors of those neglect of left-behind children in rural area,and to provide bases for the development of intervention measures.Methods 2917 students were selected as the study subjects from Changfeng county of Anhui province with cluster sampling method and were evaluated by a Parents-Child Conflict Tactics Scales and questionnaire on influential factors.Results 1694 left-behind children,accounted for 58.1％ of the total students,were surveyed in this investigation.The prevalence rates of neglect,among total children,left-behind children,non-left-behind children were 67.4％,70.2％,63.5％,respectively.The prevalence of neglect among left-behind children was higher than that among non-left-behind children (x2=14.322,P＜0.000).There were no significant associations with the neglect rate of left-behind children regarding gender or age differences.Result from multivariate logistic regression analysis indicated that the neglect among the left-behind children were associated with family dysfunction(OR values of moderate and serious family dysfunctions compared to good family function were 1.628 and 2.341,respectively)and the rate of keeping in touch with parents(OR values of sometimes and seldom keeping in touch compared to regular in touch were 1.299 and 1.844,respectively).The starting age of being left-behind(OR values of starting age that being left-behind from 6 to 10 and ≤5 years relative to starting age of left-behind ≥11 years were 0.703 and 0.630,respectively)appeared to be the protection factor to the neglect of those left-behind children.Conclusion Our findings indicated that the status of neglect among the left-behind children was serious.Prevention programs on the issue should target on a number of factors,including the characteristics of the chldren them-selves,as well as on the family of the children.

Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P＜0.01)and ROS level increased (P＜0.01).CoQ10 could improve PC12 cell viability (P＜0.01)and reduce the level of ROS (P＜0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P＜0.05),active Caspase-3 (P＜0.05)and Bax (P＜0.01)expressions,increase the expression of Bcl-2 (P＜0.01),and prevent nuclear translocation of AIF (P＜0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.

OBJECTIVETo investigate the optimal glycated haemoglobin (HbA1c) cut off points and evaluate the impact of HbA1c on diabetes and pre-diabetes in middle-aged and elderly population.

METHODSSubjects were recruited from Shanghai Changfeng Study. A total of 1973 community-based participants (age ⋝45) without known diabetes underwent oral glucose tolerance test (OGTT) by using a 75-g oral glucose load and HbA1c was measured by using high performance liquid chromatography (HPLC). Subjects were classified as normal glucose tolerance (NGT), pre-diabetes(impaired glucose regulation, IGR) and new diagnosed diabetes (NDD) per 1999 WHO criteria. Two tests are compared with receiver operating characteristic curve (ROC).

RESULTSAmong 1973 subjects, 271 (13.7%) were diagnosed as NDD and 474 (24.0%) as IGR by using OGTT. HbA1c was 5.7%±0.7% in this population. Use of 6.5% as the HbA1C cutoff point has sensitivity of 38.7% and specificity of 98.5%. We recommend 6.0% as a better cutoff value for diagnosis of diabetes in this population (AUC 0.829, 95% CI 0.798-0.860, P<0.001) with its sensitivity and specificity as 66.1% and 86.8%. For IGR, the results showed low sensitivity (44.9%) and specificity (66.7%) with an AUC of 0.571 for HbA1c when 5.8% was used as the cutoff point. Participants detected with HbA1c⋝6.0% were associated with nearly the same metabolic characteristics, including body mass index (BMI), blood pressure, lipid profile and urine albumin-creatinine ratio (uACR) compared with diabetic subjects detected by OGTT.

CONCLUSIONThe optimum HbA1c cutoff point for diabetes in our study population was lower than ADA criteria, and HbA1c may not be used to identify IGR.

Aged ; China ; Diabetes Mellitus ; diagnosis ; metabolism ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Male ; Middle Aged ; Prediabetic State ; diagnosis ; metabolism