Adolescent ; Child ; Child, Preschool ; Female ; Guillain-Barre Syndrome ; blood ; Humans ; Immunologic Factors ; blood ; Interleukin-13 ; blood ; Interleukin-18 ; blood ; Male

OBJECTIVETo verify the regulatory effects of acupuncture on exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) at stable phase.

METHODSThirty cases of COPD were randomly divided into a treatment group (16 cases) and a placebo group (14 cases). Based on specified aerobic exercise, acupuncture was applied in the treatment group and placebo acupuncture was used in the placebo group. The acupoints included Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and so on. The needle did not penetrate into the skin for the placebo group. The treatment was required for 2 to 3 times per week for totally 5 weeks. The indices of exercise tolerance, including 6-min walking distance (6-MWD), exercise time, maximum oxygen uptake (VO2max) forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximum ventilatory volume (MVV), St. George respiratory questionnaire (SGRQ) were observed in two groups before and after treatment.

RESULTS(1) Exercise tolerance: the differences of 6-MWD and exercise time were statistically significant between groups, which were more superior in the treatment group (both P<0.01); the VO2max was significantly increased after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05). (2) Pulmonary ventilation function: the differences of FEV1%, FEV1/FVC and MVV% were statistically significant between groups, which were more superior in the treatment group (P<0.05, P<0.01); (3) SGRQ: the SGRQ was significantly improved after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05).

CONCLUSIONThe acupuncture could improve the exercise tolerance in patients with chronic obstructive pulmonary disease at stable phase, and shorten the onset time of aerobic exercise. Besides, acupuncture combined with aerobic exercise could effectively improve the pulmonary function.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Exercise Tolerance ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

Objective:To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by IL-8 in vitro.Method：An in vitro incubation system was used. Superoxide anion production was determined by cytochrome C reduction. β-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and Micrococcus Lysodeikticus were as the substrates, respectively.Results:In comparison with control, musk-1 at concentration 1～100 μg*ml-1 can increase superoxide anion production by 91.7%～291%, and decrease β-glucuronidase and lysozyme release by 2.2%～58.1% and 3.9%～39.8%, respectively.Conclusion:Inhibition of lysosomel enzyme release might be considered as one of mechanisms of antiinflammatory action of musk.

AIMTo explore possible signal transmission way through which ginsenoside Rg1 protect cells from MPP(+)-induced apoptosis.

METHODSThe apoptosis of SHSY5Y induced by 1-methyl-4-phenylpyridinium (MPP+) was observed by AO-EB staining. Flow cytometry was used to quantitate the reactive oxygen species (ROS). Western Blotting was used to detect the c-jun NH2-terminal kinase (JNK) activity in SHSY5Y cells. Immunocytochemistry staining was used to detect cleaved Caspase-3 positive cells.

RESULTSMPP+ was shown to induce apoptosis in SHSY5Y cells. The percentage of apoptotic SHSY5Y cells induced by MPP+ was obviously lower in those groups pretreated with 10 mumol.L-1 Rg1 or 2.5 mmol.L-1 N-acetylcysyteine (NAC). It showed more ROS in MPP+ groups than in control. JNK activity increased with time within 72 hours in 1 mmol.L-1 MPP+ group. Simultaneously, it showed decrease of ROS, less activity of JNK and lower expression of cleaved Caspase-3 in 10 mumol.L-1 Rg1 and 2.5 mmol.L-1 NAC pretreated groups compared with groups treated with MPP+ only.

CONCLUSIONRg1 protects against MPP(+)-induced apoptosis in SHSY5Y cells and the effect might be attributed to its removal of ROS, inhibition of the activity of JNK and expression of cleaved Caspase-3.

1-Methyl-4-phenylpyridinium ; antagonists & inhibitors ; pharmacology ; Apoptosis ; drug effects ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ginsenosides ; isolation & purification ; pharmacology ; Humans ; JNK Mitogen-Activated Protein Kinases ; MAP Kinase Kinase 4 ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Neuroblastoma ; pathology ; Neuroprotective Agents ; pharmacology ; Panax ; chemistry ; Reactive Oxygen Species ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo explore bone marrow-derived mesenchymal stem cells (BMSC) mediated gene directed enzyme prodrug targeting anti-tumor therapy (GDEPT).

METHODSCYP1A2 gene was cloned from human hepatocytes by RT-PCR, and the eukaryote expression vector was constructed and transferred into Raji cells and human BMSCs via liposome. The targeted anti-tumor effect of BMSC-CYP1A2 cooperated with dacarbazine (DTIC) was measured. RT-PCR and Western blot were used to detect the expression of CYP1A2. Migration assay was detected with Transwell Plates. MTT was used to evaluate the growth inhibitory effect. Cell apoptosis was determined with Annexin V-FITC/PI staining by flow cytometry(FCM).

RESULTSThe results of FCM and differentiation induction were in line with the characteristics of BMSC. The expression of CYP1A2 was confirmed by RT-PCR and Western blot. Growth inhibition of Raji-CYP1A2 cells was increased with DTIC concentration in a dose-dependent manner (IC(50) was 1.67 mmol/L). However, BMSC was less sensitive to the cytotoxic effects of DTIC (IC(50) of 9.26 mmol/L and 7.53 mmol/L for BMSC-pcDNA3.1 and BMSC-CYP1A2 cells, respectively) than Raji cells did (IC(50) of 5.62 mmol/L and 1.67 mmol/L for Raji-pcDNA3.1 and Raji-CYP1A2 cells, respectively). BMSC migrated toward Raji cells in Transwell plate. BMSC-CYP1A2 cells mediated a bystander killing effect for CYP1A2-negative Raji cells when they were co-cultured with BMSC-CYP1A2 cells.

CONCLUSIONDTIC can be catalyzed by CYP1A2 in vitro. BMSC-based enzyme prodrug system of CYP1A2 and DTIC can induce lymphoma cell apoptosis targetedly via bystander killing effect.

Apoptosis ; drug effects ; genetics ; Bone Marrow Cells ; Cells, Cultured ; Cytochrome P-450 CYP1A2 ; genetics ; Dacarbazine ; pharmacology ; Genetic Vectors ; Humans ; Lymphoma ; pathology ; Male ; Mesenchymal Stromal Cells ; Transfection

OBJECTIVEStudying the gene expression profiling of different stage hairy root of Salvia miltiorrhiza, in order to find functional genes.

METHODThe contents of second metabolites were determined by HPLC and gene expression profiling was detected by cDNA microarray. cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine's linear RNA amplification method and subsequent enzymatic reaction. The microarrays were scanned with a ScanArray Express scanner using ScanArray 2.0 software and quantified by signal intensities of individual spots from the 16-bit TIFF images using GenePix Pro 4.0. The linear normalization method was used for data analyze. Northern blot was used to test the gene expression results obtained by microarray. Different expressed genes were sequenced and analyzed by gap4 software, and then they were analyzed with BLASTX, BLASTN, GO and KEGG.

RESULTGrowth rate and second metabolites analysis indicated that the stage from 30 d to 45 d was the growth stage, while the stage from 45 d to 60 d was the second metabolites accumulation stage. Accordingly 30 d hairy root was chosen as a reference, which was hybridized with 45 d and 60 d hairy root separately. Total 203 different expressed genes were obtained. Northern blot showed that the result was identical with the microarray result. After sequenced, there were 172 genes clustered into 114 clusters (Unigenes). Among them, 62 unigenes had known functions, 34 unigenes were hypothetical protein, 9 unigenes were homologues with no similarity and 9 unigenes were unidentified protein with low similarity. Total 67 genes were classified into cellular component ontology, molecular function ontology and biological process ontology based on GO analysis. Total 26 genes, which represented 29 metabolic-related enzymes, were located in metabolic maps based on KEGG pathway classification.

CONCLUSIONSeveral important functional genes related to second metabolite synthesis were cloned such as P450 and copalyl diphosphate synthase genes. cDNA microarray was a useful tool for functional genomics of traditional Chinese medicine.

Alkyl and Aryl Transferases ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism

OBJECTIVETo characterize the HA1 regions of hemagglutinin gene of influenza viruses (H3N2) isolated from children in Beijing from 1998 - 2004.

METHODSThe HA1 regions of hemagglutinin gene were amplified by RT-PCR from the viruses isolated and identified as A3 (H3N2) from clinical samples collected from infants and children during the peak seasons of influenza between 1998 and 2004. PCR products were sequenced or cloned into T-A vector and were analyzed after being sequenced.

RESULTSThe HA1 regions of hemagglutinin genes amplified from those isolates were 987 bp in length, encoding a protein of 329 amino acids in length. The identities of nucleotides and amino acids among these H3N2 isolates in Beijing and vaccines strains from 1998 - 2004 were 95.5% - 100.0% and 93.0% - 100.0%, respectively. The homology of the HA1 regions were related to the date of virus isolation, meaning the homology was higher among those strains isolated in nearer dates than others. Seven potential N-linked glycosylation sites in the HA1 regions located at amino acid positions 8, 22, 38, 63, 126, 165 and 285 were conserved in all the viruses analyzed. Two sites at 122 and 133 were inserted in those virus isolated after 1997, and another site at 144 appeared in those isolated after 1999. More amino acid substitutions located in the five putative antigenic sites or receptor binding sites were found more in the isolates than the isolates from previous year. Phylogenetic analysis showed new branches appeared continuously during 1998 - 2004. The strains isolated during winter in 2004 belonged to different branches, suggesting the appearance of new variants.

CONCLUSIONAmino acid substitutions continuously occurred in the HA1 regions of hemagglutinin genes in influenza virus (H3N2) isolated from children in Beijing from 1998 - 2004, which might have resulted in antigenic drift and led to the appearance of new variants.

Amino Acid Substitution ; China ; DNA, Viral ; analysis ; Gene Amplification ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; Influenza, Human ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo investigate the growth status of children under 7 years in Wuzhong City, Ningxia Hui Autonomous Region, China and its influential factors, and to provide a basis for related intervention measures.

METHODSChildren under 7 years were selected from two county-level districts in Wuzhong by stratified cluster sampling, and their growth status were evaluated by the Z score method.

RESULTSThe prevalence rates of growth retardation, underweight, and wasting were 12.58%, 5.71%, and 5.55% respectively. The height-for-age Z score, weight-for-age Z score, and weight-for-height Z scores were -0.26±2.50, 0.29±4.54. and 0.65±3.02 respectively. There were significant differences in the prevalence rate of wasting among children of different ethnic groups (P<0.05); also, there were significant differences in the prevalence rates of growth retardation and underweight among children from different regions and with different age (P<0.05). The main influential factors for growth retardation were region (OR=0.369, P<0.001), ethnic groups (OR=1.694, P=0.027), and age (OR=1.143, P=0.002). The main influential factors for underweight were region (OR=0.453, P=0.001) and age (OR=1.204，P=0.002). The main influential factor for wasting was nation (OR=1.735, P=0.024).

CONCLUSIONSIn Wuzhong, children under 7 years have poor growth status, which are related to ethnic groups, region, and age.

Body Height ; Body Weight ; Child ; Child Development ; Child, Preschool ; China ; epidemiology ; Diet ; Female ; Growth Disorders ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Logistic Models ; Male

OBJECTIVETo explore the impact of 5-fluorouracil (5-FU) on glucose metabolism and pancreatic pathology.

METHODSTwenty Wistar rats were divided into 5-FU group(n=10, chemotherapy was administered intraperitoneally to animals at a dose of 20 mg/kg daily for continuous 5 days) and control group (n=10, sodium chloride was administered intraperitoneally to animals with the same dose at the same time ). Glucose tolerance was evaluated 2 and 7 days following 5-FU treatment by serial measurement of blood glucose before and after an oral glucose load. Plasma insulin concentration was determined by radioimmunoassay. Pancreatic pathology was examined with morphological method and the ultrastructural changes of β cells were observed by transmission electron microscope.

RESULTSFasting blood glucose level was significantly higher in the 5-FU group than that in the control group [(7.6±0.9) mmol/L vs. (4.6±0.6) mmol/L at day 2; (8.9±1.0) mmol/L vs. (4.7±0.6) mmol/L at day 7, P<0.01]. Insulin releasing test indicated that the early phase insulin response to glucose load was significantly diminished in animals treated with 5-FU at day 2. Insulin level was significantly lower in the 5-FU group than that in the control group at 30 min (P<0.01). The peak secretion time of plasma insulin in 5-FU group was at 60 min, similar to the control group; and plasma insulin level decreased more slowly. Plasma insulin level was higher in 5-FU groups than in control groups on 120 min and 180 min. At day 7, Insulin level was lower in the 5-FU group than that in the control group on 60 min, and the peak secretion time of plasma insulin was delayed to 120 min. Plasma insulin level was significantly increased in 5-FU group than that in control group on 180 min(P<0.01). No gross histopathological damage to the pancreas was observed at day 2 and 7 following administration of 5-FU. The structural changes of mitochondria were mainly the quantities of secretory granule diminished at day 7 under transmission electron microscope. Dilated rough endoplasmic reticula, swollen mitochondria, and the presence of adipose drops in lysosomes were found in few cells.

CONCLUSIONS5-FU-induced hyperglycemia appears to be mediated in part by a relatively deficient insulin secretion to glucose stimulation. A relative deficiency in insulin secretion following 5-FU treatment appears to be related to β cells function impairs with islet cell ultrastructural changes induced by 5-FU.

Animals ; Blood Glucose ; drug effects ; metabolism ; Female ; Fluorouracil ; pharmacology ; Insulin ; blood ; Male ; Pancreas ; drug effects ; pathology ; Rats ; Rats, Wistar

OBJECTIVETo establish a rapid, specific and effective technique for identifying subtyping A(1), A(3) and B of influenza virus isolates and clinical specimens as well as to analyze the sequences of nucleotides and deduced amino acids of HA1 regions from isolates of influenza virus A(3) isolated from 1996 to 2002.

METHODSSix inner and outer sets of oligonucleotide primers were designed to detect, type and subtype human influenza A and B. The first two corresponding sets differentiate type A and B of matrix (M) gene while, the second two corresponding sets identify the H(1) and H(3) subtypes of type A virus HA gene. To type and subtype influenza viruses in clinical isolates, a mixture of inner primer sets specific for H(1), H(3) and B were used in a single PCR reaction tube. To detect influenza viruses in clinical specimens, a mixture of the outer primer sets were used in a single primary PCR tube, and the inner ones in a single second PCR reaction tube. Amplified products were visualized in 1.2% agrose gel containing ethidium bromide. HA1 regions of hemagglutinin of 5 field strains (H3N2) isolated from 1996 to 2002 in Beijing were amplified by RT-PCR and sequenced directly.

RESULTSThere was 100% correlation between multiplex RT-PCR and culture to type and subtype influenza viruses from clinical isolates. For typing and subtyping, 76.9%, 57.1% and 86.5% were positive for A(1), A(3) and B by multiplex nested-PCR compared within virus isolation on culture, respectively. The sequence data of HA1 of A(3) strains showed that there was a high homology of nucleotide and amino acid, and the closer the date of isolating was, the higher homology showed.

CONCLUSIONSMultiplex RT-PCR and nested-PCR for influenza viruses could provide a useful alternative to existing methods of influenza detected and identified from clinical isolate and specimens. There were certain, continuous mutations and increasing glycosylated sites which might cause the antigen drift in the A(3) strains during 1996-2002 in Beijing area.

Amino Acid Sequence ; Child ; China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza B virus ; classification ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity