OBJECTIVETo observe the effects of qingchang huashi recipe (QHR) on the dendritic cells (DCs) of experimental colitis rats, thus exploring its possible mechanisms for treating ulcerative colitis (UC).

METHODSThe UC rat model was induced by TNBS/anhydrous alcohol. Forty male Wistar rats were randomly divided into 4 groups, i.e., the normal group, the model group, the QHR group, and the Mesalazine group, 10 in each group. Since the 2nd day of modeling, corresponding medication was respectively administered to each treatment group by gastrogavage for 10 successive days. The number of DCs in the colonic mucosa was observed using iMmunohistochemical assay. The DCs ratio in the mesenteric lymph nodes, and the expressions of surface molecules MHC-II and CD86 were detected using flow cytometry.

RESULTSCompared with the model group, the number of DCs in the colonic mucosa significantly decreased, the expression of MHC-II in the mesenteric lymph nodes significantly decreased in the QHR group and the Mesalazine group, showing statistical difference (P < 0.01). There was no statistical difference between the two groups (P > 0.05). There was no statistical difference in the DCs ratios and the CD86 expression among the 4 groups (P > 0.05).

CONCLUSIONQHR could decrease the infiltration of DCs in the colonic mucosa, and suppress the activation of DCs in the mesenteric lymph nodes, which might be one of its mechanisms for treating UC.

Animals ; Colitis, Ulcerative ; drug therapy ; physiopathology ; Dendritic Cells ; cytology ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Intestinal Mucosa ; cytology ; Lymph Nodes ; cytology ; Lymphocyte Count ; Male ; Mesentery ; cytology ; Phytotherapy ; Rats ; Rats, Wistar

Objective To assess differential expression profiles of microRNAs(miRNAs) in HaCaT human keratinocytes before and after p53 gene silencing,and to make a functional analysis of target genes.Methods Lentivirus-mediated RNA interference (RNAi) was used to silence p53 gene in HaCaT cells.Total RNA was extracted using Trizol reagent.Then,miRNAs were isolated by polyethylene glycol (PEG) and subjected to fluorescent labeling using T4RNA ligase followed by hybridization to a mammalian miRNA chip.Microarrays were scanned by a GenePix 4000B microarray scanner and fluorescence ratios were determined with the GenePix Pro 6.0 software.The TargetScan software was used to predict target genes of differentially expressed miRNAs (＞2-fold difference in expression level),and the top 20 target genes with the highest enrichment score were selected for each miRNA and subjected to functional analysis and pathway analysis through the KEGG signaling database.Results Totally,53 differentially expressed miRNAs,including 12 down-regulated and 41 up-regulated miRNAs,were identified in HaCaT cells after p53 silencing as compared to those before p53 silencing.Of these 53 differentially expressed miRNAs,5 (hsa-miR-141-3p,hsa-miR-15a-5p,hsa-miR-27a-3p,hsa-miR-130b-3p,hsa-miR-19a-3p) showed a more than 200-fold increase in expression,and 4 (hiv1-miR-TAR-3p,hsa-miR-630,hsa-miR-1246,hsa-miR-1275) experienced a more than 4-fold decrease in expression in HaCaT cells after p53 silencing.Functional analysis and pathway analysis revealed that some target genes of these differentially expressed miRNAs were involved in the mitogen-activated protein kinase (MAPK) signaling pathway,metabolic pathways,and tumor invasion.Conclusion Nine miRNAs,including hsa-miR-141-3p,may be involved in p53-mediated molecular regulation.

Objective To explore the possible relationship between environmental contamination by Aspergillus and invasive aspergillosis.Methods From November 2005 to October 2006,samples were collected from the environment (air in corridors,air in wards,surfaces and tap water) twice a month,and from patients (nose,pharynx and sputum) at a liver transplantation department (LTD),neurologic surgery intensive care unit (NSICU) and central intensive care unit (CICU) in our hospital,and subjected to fungal culture.The Aspergillus density was determined in these environments.The isolates of Aspergillus flavus were genotyped by random amplification of polymorphic DNA (RAPD) assay to investigate the origin of infection.Results The mean aspergillus density was 12,10.75,0 and 20 cfu/m~3 at LTD,NSICU,CICU and corridors respectively.The five most prevalent species of aspergillus in these environments in decreasing order were Aspergillus flavus,Aspergillus fumigatus,Aspergillus niger,Aspergillus versicolor and Aspergillus clavatus.RAPD demonstrated that the genotypes ofA.flavus isolated from two patients were identical to those of the environmental strains in NSICU.The A.flavus genotypes from 3 patients in CICU were all different from those of the environment strains in CICU,but the genotypes were identical from two of the three patients.Conclusions Aspergillus contamination of different degree does exist at LTD,NSICU and CICU. The genotypes of A.flavus are identical from patients and environment in NSICU,suggesting that the clinical infection may originate from hospital environment.

OBJECTIVETo observe the therapeutic effects of beta-elemene combined DC/Dribble vaccine in treating mice with hepatic cancer, thus exploring their anti-tumor mechanisms.

METHODSDentritic cells were derived from Balb/c mice's spleen and their phenotypes were identified. Using hepatic cancer cell line BNL1MEA.7R.1 (abbreviated as BNL) originated from Balb/c mice as target cell, DC/Dribble vaccine was prepared via raising the antigen representing carrier autophagosomes (DRips in Blebs, DRibbles), which were rich in tumor antigen information. The mice previously immunized were divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The PBS was subcutaneously and intraperitoneally injected to mice in the control group. The beta-elemene was intraperitoneally injected at the daily dose of 50 mg/kg to mice in the beta-elemene group and the combined group for 7 successive days. DC/Dribble vaccine was injected into the lymph node of mice in the vaccine group and the combined group on the 1st day, and DC/Dribble vaccine was subcutaneously injected on the 3rd day and the 5th day. All the mice were sacrificed on the 10th day. Their spleens were obtained sterilely, and the suspension was incubated with or without Dribble. The cells were inoculated for 72 h. The contents of IFN-gamma in the supernatant were measured by ELISA. In addition, the spleen cells obtained from the combined group were incubated with different stimulations for 72 h, which were then divided into the control group, the DRibble group, the DC group, and the DC/Dribble vaccine group. The supernatant of cultured cells were collected and the contents of IFN-gamma were measured by ELISA. The liver tumor-bearing mouse model was established, and then the BNL bearing mice were randomly divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The treatment ways were the same as the immune ways. The tumor size and the survival period were observed in each group. On the 23rd day the mice were sacrificed. The tumor tissue was stripped and stained by HE staining. The pathomorphological manifestations of the tumor tissue were observed by light microscope.

RESULTSIn vitro detection of mice immunized previously by different ways showed that the secretion of IFN-gamma was significantly higher in the combined group than in the rest groups (P < 0.01). The secretion of IFN-gamma was significantly higher in the beta-elemene group and the vaccine group than in the control group (P < 0.01). The spleen cells could be stimulated to secrete a large amount of IFN-gamma in the vaccine group and the Dribble group (P < 0.01). When the beta-elemene was 10 microg/mL as the stimulating dose, the secretion of IFN-gamma obviously increased (P < 0.01). In vivo observation showed that the growth velocity of tumors in mice of the combined group was slowed down. There was statistical difference in the tumor area or the survival period of mice in the combined group, when compared with the other groups (P < 0.01). In HE staining, the surrounding connective tissues of the tumor were wrapped tightly and compactedly, with infiltration of a large amount of inflammatory cells.

CONCLUSIONSbeta-elemene combined DC/Dribble vaccine could induce specific immune cells to secrete secretory cells, thus exerting its anti-tumor effect. Its immunological effects might be associated with enhancing the DC antigen presenting function.

Animals ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Dendritic Cells ; drug effects ; immunology ; Female ; Liver Neoplasms ; drug therapy ; immunology ; Mice ; Mice, Inbred BALB C ; Sesquiterpenes ; pharmacology

Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Arsenicals ; therapeutic use ; Fluoresceins ; Liver Neoplasms, Experimental ; chemically induced ; drug therapy ; Male ; Oxides ; therapeutic use ; Rats

Cardiovascular Surgical Procedures ; methods ; Hemorrhage ; prevention & control ; Humans

The best absorption location of puerarin microemulsion-in-oil in intestine parva of rat and pharmacokinetic characteristics, and the pathway of absorption and conveying of puerarin microemulsion were studied. In situ rat perfusion method was used to investigate the intestinal absorption of puerarin. Through the changes of drug concentration in blocked and unblocked lymphs, to determine the pathway of absorption and conveying. Puerarin microemulsion-in-oil can be absorbed in any part of intestine, and the K(a), P(app) of every part is ileum > duodenum > jejunum > colon, and the K(a), P(app) of ileum is significantly larger than that of others. The absorption rate of different concentrations is not significantly different (P > 0.05). The puerarin transited by gastrointestinal tract, about 36.8% is absorbed by the lymphatic channels to enter the systemic circulation and 63.2% is absorbed by the non-lymphatic channels. The best part of intestine to absorb puerarin microemulsion is ileum, and it is passive transport. The pathway of conveying is lymphoid and non-lymphoid transit.
Absorption ; Animals ; Colon ; metabolism ; Duodenum ; metabolism ; Ileum ; metabolism ; Intestinal Absorption ; Isoflavones ; administration & dosage ; pharmacokinetics ; Jejunum ; metabolism ; Nanostructures ; Particle Size ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the effect of PTH gene polymorphisms on bone mineral density (BMD) at multiple skeletal sites in normal females.Methods PTH gene phenotype was determined by PCR-RFLP of restriction enzyme Bst BⅠin 596 females aged (46.3?13.7) years (20-80 years),and PCR products with or without enzymolytic site were considered as genotype B or genotype b respectively.BMDs of the anteropesterior spine (AP) and supine lateral spine (Lat) of lumbar vertebrae (L_1-L_4),femoral neck (FN),total hip (T-hip), Ward's triangle (Ward),Trochanter (Troch),forearm [radius+ulna ultradistal (RUUD) and total area of radius + ulna (RUT) ] were measured by DEXA (QDR4500A).Results (1) Hardy-Weinberg equilibrium was evident for PTH polymorphisms.The frequencies of genotype were BB 0.784,Bb 0.208,bb 0.008 and frequencies of alleles B,b were 0.888 and 0.112 respectively in 596 normal females.Frequencies of BB,Bb,bb genotypes were 0.781,0.210,and 0.009 respectively in 347 postmenopausal women and their frequencies of alleles B,b were 0.886,0.114.No significant difference was found between post- and premenopausal women in genotype frequen- cy.(2) Comparing their BMDs of AP,Lat,FN,T-hip,Ward,Troch,RUUD and RUT,there was no significant difference between BB and Bb genotypes of pre- and postmenopansal women groups.(3) Logistic regression analysis failed to show any statistical difference between normal and osteoporosis women with regard to PTH phenotype.Conclusion PTH gene polymorphism has little effect on BMD in normal females.

Objective To investigate and analyze the situation of urban and rural neglected children aged 3-6,in China,so as to provide basis for the analysis and comparison on relevant risk factors.Methods 1163 urban children aged 3-6 (with 49.6％ males and 4.5％ with minority ethnicity) were investigated from 25 cities of 14 provinces,autonomous regions and municipalities in the whole country.Multi-stage stratified cluster sampling method was used.Again,using the same sampling method,4096 rural children (of whom 50.6％ were males with 6.2％ as minorities) were chosen from 26 cities of 10 provinces or municipalities.Identification of children being neglected was based on “Child Neglect Evaluation Norms of Children Aged 3-6 Years in Urban/Rural China”.SPSS-Windows 13.0 was employed for data analysis.Scores,frequency/degrees,age,sex and types (physical,emotional,educational,safety,medical and social) of children under negligence on every group of the regions,were calculated.x2 test (Chi-Square) and Analysis of variance (ANOVA) were processed to determine the significance of their differences.Results The overall frequencies of negligence were 28.0％ and 53.7％ respectively among the urban and rural children aged 3-6,while the total degrees of negligence were 42.2 and 44.4 respectively.Significant difference was found between children from the urban and the rural areas (P＜0.05).Significant difference was also found between urban and rural children on every age group (P＜0.05).The frequencies of negligence among males were 32.6％ and 55.9％ respectively in urban and rural areas while among females,the figures appeared to be 23.7％ and 51.6％ respectively.The degrees of negligence were 42.7 and 44.6 among male while 41.8 and 44.3 among female children,in the urban or rural areas.Significant differences were found on male or female between urban and rural groups (P＜0.05).Frequencies of negligence in urban children aged 3-6 for the six types were from 5.1％ to 12.9％,with the frequency in rural areas as 13.1％-26.6％.Significant difference was found between urban and rural group for any other type (P＜0.05),in addition to the safety type.The degrees of negligence in urban children aged 3-6 for the different type were between 39.4 and 43.4,while in the rural areas as from 36.5 to 48.2,with significant difference for every type (P＜0.05).The degrees of negligence related to education,emotion,or physical strength were more serious on children from the urban than from the rural areas.The highest frequency of child negligence was seen in the single-parent families on both urban and rural groups (42.9％ and 60.0％ respectively),with no significant difference found (P＞0.05).The urban and rural children aged 3-6 were mainly involved in single item of negligence,with incidence rates as 16.5％ and 22.7％ and proportions as 58.9％ and 45.1％ respectively,despite the factors as age or sex.Conclusion There were large differences on the situation of negligence between the urban and rural children aged 3-6.The frequencies and degrees of negligence in every age group and different sex for children living in the rural areas were higher than those urban children.The frequency of negligence among boys was higher than girls for both urban and rural areas.The rural children had suffered more serious negligence than the urban children at any other type,in addition to the ‘ safety'.Both urban and rural children had the highest frequency of negligence in single-parent family,and were mainly suffered from single item of negligence.

OBJECTIVETo study the effects of androgen on the expression of aromatase cytopigment P450 (AROM) and nerve growth factor (NGF) in the brain and brain ultrastructure in neonatal rats with hypoxic-ischemic brain damage (HIBD) in order to investigate the mechanism underlying the protective effect of androgen against HIBD.

METHODSNinety-six seven-day-old Sprague-Dawley rats were randomly divided into three groups: sham-operation, HIBD and androgen treatment (n=32 each). HIBD was induced by the ligation of left common carotid artery and hypoxia exposure. The rats in the androgen treatment and the HIBD groups were injected intraperitoneally with testosterone propionate (25 mg/kg) and arachis oil respectively immediately after hypoxia-ischemia (HI). After 24 and 72 hrs and 7 and 10 days of HI, AROM and NGF expression in the cortex and the hippocampus was detected with the immunohistochemical method. The ultrastructural changes of neurons in the cortex and the hippocampus were observed under a transmission electron microscope.

RESULTSNerve cells of the HIBD group showed obvious injuries including cell organ decreasing, cellularoedema, nuclear swelling, chromatic agglutination, mitochondria decreasing and swelling, as well as an increase in apoptotic cells. Compared with the HIBD group, the nerve cells in the androgen treatment group had integrated nuclear membrane, well-distributed chromatin and abundant cell organs, and less cell apoptosis and increased axon regeneration. There was a positive expression of NGF and AROM in the brain cortex and the hippocampus in the HIBD group 24 hrs after HI. The expression of NGF and AROM increased significantly 72 hrs after HI, peaked 7 days after HI and then began to decrease but remained at a higher level than that in the sham-operation group 10 days after HI. The NGF and AROM expression in the cortex and the hippocampus in the androgen treatment group was significantly higher than that in the sham-operation and the HIBD groups 72 hrs, and 7 and 10 days after HI.

CONCLUSIONSAndrogen treatment can promote axon regeneration and morphous recovery of neurons and decrease neural apoptosis in neonatal rats with HIBD. The neuroprotection of androgen is produced possibly through an increase in the expression of NGF and AROM in the brain.

Androgens ; therapeutic use ; Animals ; Animals, Newborn ; Aromatase ; analysis ; Brain ; enzymology ; Female ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; pathology ; Immunohistochemistry ; Male ; Nerve Growth Factor ; analysis ; Neurons ; ultrastructure ; Rats ; Rats, Sprague-Dawley