Aim To investigate the effect of dihydromyricetin on canine carotid artery and the underlying mechanism.Methods The in vitro isometric tension measurement technique was employed to investigate the effect of dihydromyricetin on canine carotid artery rings.Laser scanning confocal microscope technique was used to measure the dynamic change of intracellular calcium concentration in single VSMC.Results Dihydromyricetin(1～300 ?mol?L-1)caused a concentration-dependent contraction of both endothelium-intact and endothelium-denuded rings.This constrictive effect was attenuated in Ca2+-free solution(P

This study is to determine the content of three alkaloids and establish the HPLC fingerprint of "Jianlian" Nelumbinis Plumula. The HPLC method of content determination was as follows: Thermo C18 (4. 6 mm x 250 mm, 5 μm) was conducted with acetonitrile-sodium dodecyl sulfonate solution-acetic acid (56: 43: 1) at a flow rate of 1.0 mL x min(-1). The monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. The method of HPLC fingerprint was as follows: Agilent ZORBAX SB-Aq C18 (4.6 mm x 250 mm, 5 μm) was conducted with gradient elution of methanol and water at a flow rate of 0.8 mL x min(-1), the monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. Similarities evaluation and hierarchical clustering analysis were applied to demonstrate the variability of 12 batches of "Jianlian" Nelumbinis Plumula samples. The results demonstrated that 11 batches showed good similarity on chemical constituents. The method could well display the chemical information of "Jianlian" Nelumbinis Plumula. It was simple, reliable and could be used for the chemical quality control of "Jianlian" Nelumbinis Plumula.
Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Nelumbo ; chemistry ; Quality Control ; Seeds ; chemistry

OBJECTIVE: This study aimed at examining the expression of Beclin1 and LC3 in the skin of rat after contusive injury and at observing the process of autophagy in response to injury or trauma. Their significance in diagnosis of trauma and application in estimation of the post injury time interval were also investigated. METHODS: Immunohistochemistry and imaging analysis were used to examine the expression level of Beclinl and LC3 at the injury site of the rat skin at different post-traumatic times. RESULTS: Expression of Beclin1 and LC3 in normal rat skin or immediately after injury maintained at a very low level, but their level started to elevate 6 hours after injury and showed increasing strong expression at post injury time of day 3, day 5 and day 7. CONCLUSION Over-expression of Beclin1 and LC3 as well as increased autophagy, a response to injury, can be observed in the rat skin at the injury site. The expression level of Beclin1 and LC3 starts to increase 6 hours and shows a linear increase up to 7 days after injury. Beclin1 and LC3 may serve as a marker for estimation of the post injury time interval.
Animals ; Apoptosis Regulatory Proteins/metabolism* ; Autophagy ; Beclin-1 ; Contusions/pathology* ; Forensic Medicine ; Immunohistochemistry ; Male ; Microtubule-Associated Proteins/metabolism* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Skin/pathology* ; Staining and Labeling ; Time Factors ; Wounds and Injuries/pathology*

OBJECTIVETo study the risk factors and abusers' characteristics on childhood sexual abuses among female students to provide reference data for preventive measures on child sexual abuse.

METHODSAnnonymous questionnaire survey of 701 adult female students in a college about their personal experience on childhood sexual abuses, their home conditions and abusers' characteristics.

RESULTSA total number of 701 female students were surveyed. 115 (22.11%) experienced non-physical contact sexual abuses before the age of 18, including 26 (3.71%) who suffered from attempted and 8 (1.14%) were forced with genital or anal sexual intercourse. 117 (16.69%) said they had suffered from sexual abuse before the age of 16. Most abusers were males (99.4%) and only a small percentage of them (3.2%) used violence. Most non-physical contact sexual abuses were from strangers (78.7%), while 71.3% of physical contact abuses were from acquaintances including 12.5% of them were teachers, 17.5% were neighbors and 21.3% were relatives. The risk factors of child sexual abuses were: frequent use of violence, bad hobbies and poor health conditions of parents, tight family relationships, and living in remarried families. Having well-educated fathers was a preventive factor.

CONCLUSIONStudies on childhood sexual abuses among females revealed that the incidents were not rare, and family environment played an important role.

Adult ; Child ; Child Abuse, Sexual ; statistics & numerical data ; China ; Family Relations ; Female ; Humans ; Risk Factors ; Students ; statistics & numerical data ; Surveys and Questionnaires ; Universities

OBJECTIVETo probe into the relationship between hypertension and hemodynamic score of cerebral vascular accumulative function by though studying the history of hypertension and level of blood pressure.

METHODSThe database of blood pressure and cerebral vascular hemodynamic indexes (CVHI) were from baseline data and measurement in a cohort. 18,512 participants who met the included criteria were enlisted in the study. The function scores were estimated using uniform methods according to CVHI. Participants were grouped by age, years of having hypertension and level of blood pressure. Differences of the function score between different groups were compared.

RESULTSThe function score in hypertension group was significantly lower than that of normal tension group, which decreased by 13.3 points. Within 5 years of hypertension history, the function score decreased as course of hypertension prolonged. The same trend was found between the score and level of both systolic and diastolic blood pressure. When systolic blood pressure elevated to 140 mmHg or diastolic blood pressure elevated to 95 mmHg, the function score was below 75 points.

CONCLUSIONThere were significant relationship between decrease of the function scores and medical history, course of hypertension as well as level of blood pressure.

Adult ; Age of Onset ; Aged ; Blood Pressure ; physiology ; Cerebrovascular Circulation ; physiology ; China ; epidemiology ; Cohort Studies ; Female ; Hemodynamics ; physiology ; Humans ; Hypertension ; epidemiology ; physiopathology ; Male ; Middle Aged

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) and determine its physical and chemical characters, recombinant NDPK-A producing E. coli was cultured in 80L fermentor under high cell density culture (HCDC) conditions. The harvested cells were treated with high pressure to break the cell up, tangential-flow microfiltration to remove the bacteria debris and ultrafiltration to concentrate the filtered solution containing target protein. The crude NDPK-A was purified by ion exchange chromatography with DEAE Sepharose Fast Flow, affinity chromatography with Cibarcron Blue 3GA Sepharose CL-4B and gel filtration with Sephadex G-100. The purity of rhNDPK-A was analyzed with SDS-PAGE and RP-HPLC. The Enzymatic activity was determined with RP-HPLC. The molecular weight (MW) was measured with matrix assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS). The N-terminal residue was sequenced with Edman method. The apparent molecular weight of rhNDPK-A in solution was determined with multiangle laser light-scattering method (MALS). It was found that the purity of rhNDPK-A was 97.3% with SDS-PAGE method and 99.2% with RP-HPLC method. The specific enzymatic activity was (900 +/- 100) u/mg. The molecular weight was 17017, which was 132 less than the calculated value according to the amino acid sequence of NDPK-A. The sequencing result of rhNDPK-A revealed that its N-terminal residue was Ala, which was the second residue on N-terminal of native NDPK-A. The calculated MW of N-terminal deleted rhNDPK-A was 17017, exactly equal to the experimental value. The result of apparent MW determination revealed that rhNDPK-A formed homohexamer in solution with a MW of 102kD. These results suggested that rhNDPK-A possessed character identical to its native counterpart of assembling into hexamer. Confirming the identity of rhNDPK-A to its native counterpart provided a good foundation for drug development and mechanism study of NDPK-A.
Amino Acid Sequence ; Humans ; Molecular Sequence Data ; Molecular Weight ; NM23 Nucleoside Diphosphate Kinases ; chemistry ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; isolation & purification ; Scattering, Radiation ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo evaluate the angiogenic effect of the Xiongshao capsule (XSC) in human umbilical vein endothelial cells (HUVEC), and to investigate the possible molecular mechanisms mediating its biological effect.

METHODSSerum pharmacology was applied in this study, in which different doses of XSC were administrated to rats orally and then XSC-containing serum (XSC-S) was collected for the following in vitro experiments. The viability of HUVEC was determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cell density was observed via phase-contrast microscopy. Fluorescence-activated cell sorting analysis with propidium iodide staining was performed to determine cell cycle phase. Cell migration was determined by wound-healing method. Capillary tube formation by HUVEC was examined using ECMatrix gel-based assay. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression levels were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbant assay (ELISA) analyses.

RESULTSXSC-S dose-dependently stimulated proliferation of HUVEC by promoting the cell cycle G1 to S progression. In addition, XSC-S treatment dramatically increased the migration and capillary tube formation of HUVEC in a dose-dependent manner. Moreover, XSC-S enhanced the expression of VEGF and bFGF at both mRNA and protein levels.

CONCLUSIONXSC can promote several features of angiogenesis in endothelial cells through up-regulating the expression of bFGF and VEGF, suggesting that XSC may be a potential novel therapeutic agent for the treatment of ischemic heart diseases.

Animals ; Capsules ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Collagen ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Laminin ; pharmacology ; Male ; Neovascularization, Physiologic ; drug effects ; genetics ; Proteoglycans ; pharmacology ; Rats ; Rats, Sprague-Dawley ; S Phase ; drug effects ; Up-Regulation ; drug effects ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

To assay the influence of dendritic cells(DCs)on the function of anti-infective immunity to Penicillium marneffei.DCs were generated from peripheral blood mononuclear cells(PBMC)and pulsed with Penicillium marneffei yeasts.DCs morphology was observed by the inverted microscope and cell surface markers of DCs were analyzed by flow cytometry.The concentrations of IL-12p70 were detected by ELISA. Mixed lymphocyte reaction was performed to assay the proliferation of T cells.The mRNA of CCR7 and CXCR4 were detected by the Real-time PCR quantifications.The acquired DCs exhibited irregular appearance and numerous long dendrites under light microscope.DCs and Penicillium marneffei yeasts were co-cultured for 24 h,numerous yeasts were observed inside the cells;an enhanced expression of the cell sur-face markers CD86、CD83、HLA-DR and CD40 were demonstrated;the expression of CCR7 and CXCR4 mRNA were also increased;the improved proliferation of T cells were observed in the mixed lymphocyte reaction.Yeasts-pulsed DCs secreted more IL-12p70 than that of non-pulsed,but less than that of LPS-pulsed DCs.DCs can engulf the Penicillium marneffei yeasts.When pulsed with Penicillium marneffei yeasts,DCs improved their expression of the co-stimulatory molecules and chemokine receptor CCR7、CXCR4,enhanced their capacity to process antigen.DCs play an important role in host defense against Penicillium marneffei infection.But the low level of the IL-12p70 production may lead to deficiency in the cell-mediated immunity against Penicillium marneffei.

Objective:To find out the difference between children with attention-deficit/hyperactivity disorder and normal control,and the difference between boys and girls with ADHD in resting-state.Methods:There were 38 children with ADHD in this study including 31 boys and 7 girls,meanwhile,42 normal controls whose age and gen der were matched with children with ADHD were involved too,including 29 boys and 13 girls.Total scores of ADHD rating scale (ADHD-RS) were used to measure the severity of symptom for both ADHD and normal control.Magnetic resonance imaging was used to collect the resting-state functional magnetic resonance imaging data,and the fractional amplitude of low frequency fluctuation (fALFF) was measured for each child.Analysis of covariance (ANCOVA) was used to find out the difference between the group and the sex.Results:Children with ADHD showed a higher fALFF in the right medial superior frontal gyrus and right supplementary motor area,and a lower fALFF in the left putamen,bilateral cerebellum and bilateral precuneus than controls (all corrected P ＜0.05).ADHD boys showed a lower fALFF in the left cerebellum and a higher fALFF in the bilateral medial superior frontal gyrus than boys without ADHD (all corrected P ＜0.05).ADHD girls showed a higher fALFF in the bilateral cerebellum and in the thalamus than girls without ADHD,and a lower fALFF in the insula than girls without ADHD (all corrected P ＜ 0.05).There was a negative relationship between the fALFF of the left cerebellum of ADHD girls and the scores of inattention symptoms.Conclusion:The fALFF difference exist between ADHD and normal controls,and between boys and girls with ADHD.

OBJECTIVEA comparative proteomic approach was used to identify and analyze proteins relevant to metastasis of hepatocellular carcinoma (HCC).

METHODSProteins extracted from 12 liver tumor tissue specimens (6 with metastases and 6 without) were separated by two-dimensional gel electrophoresis (2-DE). Comparative analyses of 2-DE protein patterns between the two groups were done using computerized image analysis. Selected proteins exhibiting statistically significant alternations were identified by mass spectrometry. Immunohistochemistry, Western blotting and RT-PCR were performed to examine the expressions of the candidate proteins.

RESULTS16 proteins including HSP27, S100A11, CK18 were identified using mass spectrometry, which were related to cell mobility, signal transduction, and energy metabolism respectively. Of these, HSP27 was found to be uniquely over-expressed in 2-DE maps of all metastatic HCCs when compared to the non-metastatic HCC tissues. Immunohistochemistry and Western blotting of HCC tissues confirmed this difference while RT-PCR did not.

CONCLUSIONThere are different proteins working together that affect the metastasis of HCCs. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets to the metastatic phenotype of HCC. The role of HSP27 in HCC metastasis warrants further investigation.

Carcinoma, Hepatocellular ; chemistry ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Gene Expression Regulation, Neoplastic ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins ; analysis ; Humans ; Liver Neoplasms ; chemistry ; pathology ; Mass Spectrometry ; Neoplasm Proteins ; analysis ; Proteome ; analysis ; S100 Proteins ; analysis