Objective To study the changes of flow parameters of superior mesenteric artery (SMA) in children with abdominal type Henoch-Schonlein purpura (HSP) using color Doppler ultrasound.Methods Ten children with abdominal type HSP and 17 controls were included in present study.The blood flow parameters of SMA[including peak velocity(PV),end-diastole velocity(EDV),resistant index(RI)]were measured at acute and recovery stage separately.Statistical analysis was conducted among groups.Results PV were (41.57±8.02)cm/s,(33.38±7.44)cm/s and (35.34±9.73)cm/s in acute stage,recovery stage and control group,respectively.There was no statistically significant difference among groups(F=2.471,P=0.10).EDV were(7.63±4.28)cm/s,(4.23±2.57)cm/s and (3.77±0.87) cm/s in acute stage,recovery stage and control group,respectively.There was significantly significant differences between acute stage group and other two groups(t=0.066,P=0.025;t=0.059,P=0.003).RI were (0.85±0.17),(1.00±0.15) and (1.04±0.13) in acute stage,recovery stage and control group,respectively.Also there was significantly significant differences between acute stage group and other two groups(t=1.391,P=0.020;t=1.239,P=0.026).Conclusion For abdominal type HSP in children,the changes of PV,EDV and RI of SMA were significant,which may help us determine the stage of disease.

ObjectiveTo observe the effect of comprehensive therapy on forearm extensor myotenositis.Methods72 cases were divided into two groups: a control group of 36 cases who were given routine treatment,and an experiment group of 36 cases who were given thermotherapy,computerized medium-frequency electrotherapy,physiotherapy,and ADL instruction,etc.After two courses,a simple grading score(for forearms) was used to assess the effect.ResultsOf the control group,22 cases were cured,10 remarkably effective,4 effective;of the experiment,30 cured,4 remarkably effective,2 effective(u=2.04, P<0.05).The difference of average score for forearms before and after the treatment were(6.58±3.17) points for the control and(8.19±3.55) for the experiment(t=2.03,P<0.05).The average days of cure were(5.60±2.54) d for the experiment group,shorter than those for the control(7.00±2.27) d(t=2.05,P<0.05).ConclusionComprehensive therapy is effective on forearm extensor myotenositis.

OBJECTIVETo investigate the transduction ability of PEP-1-CAT fusion protein into human umbilical vein endothelial cell (HUVECs) and the effects on hydrogen-peroxide (H2O2)-induced oxidative stress injury in these cells.

METHODSWith the use of TA-cloning program and isocaudamer technique, the pET15b-PEP-1-CAT of prokaryotic expression plasmid was successfully constructed. The recombinant plasmid was transformed into E.coli BL21 (DE3) and the protein expression was induced by IPTG. The recombinant protein has an N-terminal His-tag which could be used to purify the target protein by affinity chromatography on a Ni2+-NTA-resin column. The fusion protein PEP-1-CAT was prepared and confirmed by specific enzyme activity in vitro. The purified PEP-1-CAT fusion protein was added on cultured HUVECs in vitro. The transduction ability of PEP-1-CAT fusion protein into cells was analyzed by Western blot and specific enzyme activity. The cells were treated with H2O2 (0.5 mmol/L) alone and in combination with PEP-1-CAT fusion protein for 4 h. Then, the cell viability, lactate dehydrogenase (LDH) and malondialdehyde (MDA) contents were measured.

RESULTSThe PEP-1-CAT fusion protein could be transduced into the cultured HUVECs in a dose- and time-dependent manner and be stable for at least 48 h. After H2O2 administration, cell viability was significantly reduced compared with control group (37.23%+/-5.68% vs. 100%, P<0.05), while LDH leakage (849.3 U/L+/-95.1 U/L) and MDA (8.23 nmol/L+/-1.58 nmol/L) content were significantly higher than that in control group (540.6 U/L+/-65.7 U/L and 2.46 nmol/L+/-1.42 nmol/L, respectively, all P<0.05). Preincubation with PEP-1-CAT proteins at various concentrations (0.25-2 micromol/L) significantly attenuated H2O2-induced cell injury.

CONCLUSIONThe PEP-1-CAT fusion protein could efficiently penetrate HUVECs and the transduced protein could attenuate cellular oxidative stress injury induced by H2O2. The PEP-1-CAT fusion protein might be a new strategy for preventing and treating oxidative stress induced diseases.

Catalase ; metabolism ; Cells, Cultured ; Cysteamine ; analogs & derivatives ; metabolism ; Endothelial Cells ; metabolism ; Humans ; Hydrogen Peroxide ; Oxidative Stress ; physiology ; Peptides ; metabolism ; Umbilical Veins ; cytology

OBJECTIVETo construct the prokaryotic expression plasmid pET15b-PEP-1-CAT to obtain purified fusion protein of PEP-1-CAT.

METHODSUsing pfu DNA polymerase, the full-length human catalase cDNA was amplified by PCR from pZeoSV2(+)-CAT plasmid, and the PCR product was added with "A" using Taq DNA polymerase. The purified product of CAT cDNA with the base A at its 3' end was ligated with pGEM-T Easy vector and transformed into DH5alpha. The correct recombinant was identified by PCR and Sal I/Bgl II digestion and named as pGEM-T-CAT. Two oligonucleotides were synthesized and annealed to generate a double-stranded oligonucleotide encoding the PEP-1 peptide, which was directly ligated into Nde I/Xho I-digested pET15b. The recombinant plasmid was identified by double-enzyme digestion and named as pET15b-PEP-1. pET15b-PEP-1 and pGEM-T-CAT were further digested by Xho I/BamH I and Sal I/Bgl II, respectively. The purified linear fragment of pET15b-PEP-1 and CAT cDNA fragment were ligated using two pairs of isocaudarners possessing different recognition sequences but producing compatible cohesive ends. The clone with the expected insert was selected using Xho I restriction analysis followed by sequence analysis. The recombinant plasmid was transformed into E. coli BL21(DE3) which was induced by IPTG. The recombinant protein possessed an N-terminal His-tag sequence which could be used to purify the target protein by affinity chromatography on a Ni(2+)-NTA-resin column. The fusion protein PEP-1-CAT was produced and confirmed by specific enzyme activity in vitro.

RESULTSSequence analysis showed that the PEP-1 and the human CAT cDNA sequence of pET15b- PEP-1-CAT had identical sequence with designed PEP-1 peptide and human catalase cDNA sequence in GenBank (accession No. AY028632), respectively. SDS-PAGE and Western blotting confirmed successful expression and purification of PEP-1-CAT fusion protein with specific activity of 77.15 U/g.

CONCLUSIONThe prokaryotic expression plasmid pET15b-PEP-1-CAT has been constructed successfully, and the successful expression and purification of PEP-1-CAT provides a basis for prevention and therapy of various disorders related to oxidative stress.

Base Sequence ; Blotting, Western ; Catalase ; genetics ; metabolism ; Chromatography, Affinity ; Cloning, Molecular ; Cysteamine ; analogs & derivatives ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Molecular Sequence Data ; Peptides ; genetics ; metabolism ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism

OBJECTIVETo evaluate the androgenic and anti-androgenic effects of GH (growth hormone) transgenic carp in male rats.

METHODSHershberger assay was carried out in castrated male SD rats aged 4-5 weeks. Testosterone propionate (TP) (0.4 mg/kg BW) was administrated for a positive control, GH transgenic carp (3.0 g/kg BW)+TP (0.4 mg/kg BW), parental carp (3.0 g/kg BW) + TP (0.4 mg/kg BW), and flutamide (Flu) (3.0 g/kg BW) were used for negative controls, and vehicle was administered orally for a blank control. All groups were administrated for 10 consecutive days. At the end of the test, animals were anesthetized, then weights of accessory sex organ were measured. Serum testosterone (T), luteinizing hormone (LH), and Follicle-Stimulating Hormone (FSH) levels were detected.

RESULTSThe weights ratios of the accessory sex organs and body weights showed no significant differences between the solvent control and the GH transgenic carp-treated groups. Serum concentrations of FSH, LH, and T of the rats treated with GH transgenic carp + TP showed no significant changes, compared with those treated with TP only.

CONCLUSIONGH transgenic carp does not have any androgenic agonist or antagonist properties in vivo screening tests.

Animals ; Animals, Genetically Modified ; Carps ; genetics ; Follicle Stimulating Hormone ; blood ; Genitalia, Male ; drug effects ; Growth Hormone ; genetics ; metabolism ; pharmacology ; Luteinizing Hormone ; blood ; Male ; Rats ; Testosterone ; blood

OBJECTIVESTo investigate the improvement of specific immune responses induced by plasmid coexpressing hepatitis B surface antigen (HBsAg) and granulocyte-macrophage colony stimulating factor (GM-CSF).

METHODSAll Balb/c (H-2d) mice were immunized with pGM-CSF/S, pS/GM-CSF, pS or control plasmids. 4 weeks later, anti-HBs titer and the levels of IL-2, IL-4 and IFN-gamma in the supernatant of splenocytes were detected using enzyme- linked immunosorbent assay (ELISA), and HBsAg-specific cytotoxic T lymphocytes (CTL) activity was measured with a 51Cr release assay, using P815/S transfectants as target cells.

RESULTSThe anti-HBs antibody titers in the serum, the levels of IL-2 and IFN-gamma, and the CTL activity in pcDNA3.1-GM-CSF-S immuned mice were higher than those in PcDNA3.1-S immunized mice (F=4.176, P<0.01; F=31.188, P<0.01; F=31.796, P<0.01; F

CONCLUSIONIt will improve the specific immune responses induced by HBsAg DNA vaccine after it is binded to the gene of GM-CSF.

Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; genetics ; immunology ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Interferon-gamma ; biosynthesis ; Interleukin-2 ; biosynthesis ; Interleukin-4 ; biosynthesis ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; immunology

Objective To investigate the distribution of traditional Chinese medicine(TCM)constitution types in patients with perimenopause metabolic syndrome in Guangzhou based on the Stages of Reproductive Aging Workshop+10(STRAW+10),so as to provide a theoretical basis for TCM constitution regulation for patients with PMS.Methods According to the diagnostic criteria of metabolic syndrome,a total of 90 patients with PMS were included.Based on the STRAW+10 staging criteria,the PMS patients were divided into early perimenopause group(-2 phase of STRAW+10,49 cases),late perimenopause group(-1 phase of STRAW+10,24cases),and early postmenopausal group(+la phase of STRAW+10,17 cases).Traditional Chinese Medicine Constitution Classification and Distinguishing Criteria were used to identify the TCM constitution types of all the subjects.At the same time,the Self-Rating Anxiety Scale(SAS)and Self-Rating Depression Scale(SDS)were used for scoring anxiety and depression.The distribution of TCM constitution types in patients with different STRAW+10 stages was analyzed,and the SAS and SDS scores of patients with different STRAW+10 stages were compared.Results(1)The primary TCM constitution types in the early perimenopause group(-2 phase of STRAW+10)were yang deficiency constitution(14 cases,29.79％),balanced constitution(10 cases,21.28％),yin deficiency constitution(six cases,12.76％)and blood stasis constitution(six cases,12.76％).In the late perimenopause group(-1 phase of STRAW+10),the primary TCM constitution types were yang deficiency constitution(six cases,25.00％),balanced constitution(four cases,16.66％),blood stasis constitution(four cases,16.66％),qi deficiency constitution(three cases,12.50％)and phlegm-damp constitution(three cases,12.50％).In the early postmenopausal group(+la phase of STRAW+10),the primary TCM constitution types were yang deficiency constitution(seven cases,46.67％),balanced constitution(two cases,13.33％),phlegm-damp constitution(two cases,13.33％),blood stasis constitution(two cases,13.33％),qi deficiency constitution(one case,6.67％)and qi stagnation constitution(one case,6.67％).(2)The SAS score and SDS score in the early perimenopause group(-2 phase of STRAW+10)were(34.55±7.46)points and(35.55±10.61)points,respectively,which were higher than those in the late perimenopause group(-1 phase of STRAW+10)[(33.83±7.73)points and(35.46±11.35)points,respectively]and in the early postmenopausal group(+la phase of STRAW+10)[(35.65±8.67)points and(36.59±12.07)points,respectively].All of the scores were higher than the overall average level.Conclusion The TCM constitution of patients with perimenopause metabolic syndrome is predominated by yang deficiency constitution.The percentage of the balanced constitution gradually decreases with the progression of STRAW+10 staging,and the biased constitutions gradually develop from yin deficiency constitution and blood stasis constitution into qi deficiency constitution and phlegm-damp constitution,and then into phlegm-damp constitution and blood stasis constitution again.With the progression of the stage,the deficiency in the fundamental of the PMS patients becomes more deficient and the pathogens of excess in the incidental also grow.

OBJECTIVETo investigate the penetrating ability of fusion protein PEP-1-EGFP with human umbilical vein endothelial cells.

METHODSTwo prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E. coli BL21 (DE3) to express EGFP and fusion protein PEP-1-EGFP, respectively. The expressed EGFP and PEP-1-EGFP were purified with Ni(2+) -resin affinity chromatography, and their capabilities of transduction into human umbilical vein endothelial cells were evaluated. The time- and dose-dependent transduction of the fusion protein PEP-1-EGFP and its stability in the human umbilical vein endothelial cells were observed. The toxicity of the fusion protein PEP-1-EGFP was detected by MTT method.

RESULTSEGFP failed to be transduced into human umbilical vein endothelial cells, whereas PEP-1-EGFP fusion protein was transduced into cells shortly in 5 minutes. Its transduction was time- and dose-dependent and the fluorescence in the cells were detected even 27 hours later. No cytotoxicity of the fusion protein PEP-1-EGFP to human umbilical vein endothelial cells was detected even when the dose reached up to 200 micromol/L.

CONCLUSIONPEP-1-EGFP fusion protein can efficiently transduce the target protein into human umbilical vein endothelial cells, which provides a basis for future researches on the transduction of antioxidant enzymes mediated by the cell-penetrating peptide, PEP-1, in ischemia-reperfusion injury therapy.

Cells, Cultured ; Cysteamine ; analogs & derivatives ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Green Fluorescent Proteins ; metabolism ; Humans ; Peptides ; metabolism ; Protein Transport ; Recombinant Fusion Proteins ; metabolism ; toxicity ; Umbilical Veins ; cytology

OBJECTIVETo investigate the frequency of CYP2C19 polymorphisms involved in clopidogrel metabolism in Fujian Han population.

METHODSFrequencies of CYP2C19* 2, CYP2C19*3 and CYP2C19*17 in 1001 unrelated Fujian Han volunteers were determined with polymerase chain reaction-restriction fragment length polymorphism and direct sequencing method.

RESULTSThe frequencies of CYP2C19*2, *3 and *17 were 32.4%, 5.8% and 0.4%, respectively. According to genotyping results, intermediate metabolizers (CYP2C19 *1/*2 or *1/*3) and poor metabolizers (CYP2C19 *2/*2 and *2/*3) respectively accounted for 47.95% and 13.99% of all subjects. Above frequencies were similar to those of Japan, Korea, Singapore, Malaysia, Thailand and Chinese Dai, Mongolian,Li and Hui ethnics (P>0.05), but were significantly different from those of Chinese Kazakh and Uygur ethnics, and people from Iran, Russia, Italy, Poland, Norway, Canada native Indians, Bolivia, Egypt or Tanzania (P<0.05).

CONCLUSIONEthnic/regional diversity exist with regard to the prevalence of CYP2C19 polymorphisms. No significant difference were found between Fujian Han Chinese and Dai, Mongolian, Li and Hui from China or other populations from East and Southeast Asia, but higher frequencies of intermediate metabolizers and poor metabolizers compared with populations of Kazakh and Uygur in China, and people from Europe, South America and Africa.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aryl Hydrocarbon Hydroxylases ; genetics ; China ; Cytochrome P-450 CYP2C19 ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Ticlopidine ; analogs & derivatives ; metabolism ; Young Adult

OBJECTIVETo study the effect of maxillary sinus floor elevation by the Frialit-2 Bone Condenser for implantation.

METHODS11 patients underwent sinus floor lift by The Frialit-2 Bone Condenser and were inserted 14 implants. The time of following up was 10 - 21 months.

RESULTSThere were no implant loose or lost, no clinical complaint of maxillary sinus area, and X-ray exam showed well osseointegration.

CONCLUSIONSThe Frialit-2 bone condenser can be used for lifting sinus floor, and the sinus elevation without lateral access allows the insertion of implants with no additional surgical stress for the patients.

Alveolar Process ; surgery ; Bone Transplantation ; Dental Implantation ; methods ; Dental Implants, Single-Tooth ; Female ; Follow-Up Studies ; Humans ; Male ; Maxillary Sinus ; surgery