Objective: To study the chemical constituents from the roots of Rubus parvifolius. Methods: The chemical constituents were extracted and purified by silica gel column chromatography. NMR spectra were used for structural identification. Results: Phytochemical study on the roots of R. parvifolius led to the isolation of one ceramide (1), two anthraquinones ( 2 and 3), four triterpenoids (4-7), two flavonoids (8 and 9), one fatty acid ester (10), and two sterols (11 and 12). Conclusion: Compound 1 is isolated from the plants of family Rosaceae for the first time, and compounds 2-5 are isolated from genus Rubus for the first time. Though R. parvifolius shares the same major chemical types (triterpenoid, flavonoid, and anthraquinone) with those of R. alceaefolius, a substituent of R. parvifolius, their individual constituents are different. In addition, R. parvifolius contains ceramide (1) with high concentration, while caffeoylquinic acid reported in R. alceaefolius has not been found in R. parvifolius. Furthermore, the results from our phytochemical study are consistent with the DNA phylogenic relationship between R. parvifolius and R. alceaefolius (two separated subgenera), suggesting that the substitution of the former species with the latter one in folk medicine might not be suitable.

Objective To explore the role of tissue inhibitor of metalloproteinase-1(TIMP-1) during renal senescence by using human TIMP-1 transgenic mice.Methods Renal histological changes of wild type mice and transgenic mice at the age of 3,12,24 months were observed by periodic acid-schiff(PAS)staining of paraffin sections.The numbers of F4/80 positive cells were detected by immunofluoreseence.The protein expressions of TIMP-1,TIMP-2,matrix metalloproteinase(MMP)-9,MMP-2,intercellular adhesion molecule-1(ICAM-1),transforming growth factor?1(TGF-?1),collagenⅢand collagenⅣwere detected by Western blot.The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively.Results Focal renal fibrosis was found in two genotypes with aging.At the age of 24 months,compared with wild type,in kidneys of transgenic type,the expressions and activities of gelatinases were dowregulated (MMP-2:2.08?0.20 vs.3.39?0.43;MMP-9:4.02?0.82 vs.6.72?1.40,all P＜0.05);the expressions of collagenⅢ,collagenⅣ,ICAM-1,and TGF-?1 were upragulated(0.72+0.11 vs.0.57?0.09;0.84?0.13 vs.0.6?0.11,0.72?0.12 vs.0.53?0.07; 0.69?0.12 vs.0.45?0.09,all P＜0.05),and the numbers of F4/80 positive cells were increased (18.8?4.4 vs.12.7?3.6,P＜0.05)with the upregulated expression and activity of TIMP-1(1.10?0.18 vs.0.62?0.09;50.75?7.25 vs.20.64?3.50,P＜0.05).Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.

Objective To examine how stroke affects muscle coordination and whether muscle function will be improved after rehabilitation.Methods Ten stroke patients with mild hemiparesis and six age-and sex-matched controls were investigated at baseline.Velocity-encoded phase-contrast magnetic resonance imaging (VE-PC MRI) and surface eleetromyography (sEMG) were performed to evaluate muscle coordination of thigh muscles during knee extension and flexion and the effect of rehabilitation. Results Using VE-PC MRI,we found that the peak velocity of rectus femoris was lower in the affected limb (P

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

OBJECTIVETo investigate the acting mechanism of Buxu Huayu Qutan Decoction (BHQD) for impacting the anti-oxidative capacity in aged patients with stable angina pectoris of coronary heart disease (AP-CHD).

METHODSForty patients of AP-CHD, Chinese medicine diagnosed as Xiong-bi, were equally assigned to the treatment group and the control group. Superoxide dismutase (SOD) activity and lipid peroxide (LPO) contents in blood, and mRNA expression of manganese-containing superoxide dismutase (MnSOD) in peripheral mononuclear cells were determined before and after treatment by biochemical and molecular biologic techniques.

RESULTSNo significant change of plasma SOD and LPO was found in the control group after treatment (P >0.05), while the plasma SOD activity increased and LPO content lowered in the treatment group significantly (P<0.01). Moreover, mRNA expression of MnSOD in the treatment group after treatment was obviously higher than that in the control group (P <0.01).

CONCLUSIONSThe acting mechanism of BHQD for AP-CHD treatment might partially due to its effects in inducing gene expression of MnSOD in mononuclear cells, enhancing SOD activity, decreasing LPO content, maintaining oxidation/antioxidation equilibrium in myocardial cells, blocking the chain reaction of lipid peroxidation, intervening the production and enhancing the scavenging of oxygen free radicals.

Aged ; Angina Pectoris ; drug therapy ; metabolism ; Coronary Disease ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Lipid Peroxidation ; Lipid Peroxides ; blood ; Male ; Middle Aged ; Oxidative Stress ; Phytotherapy ; Superoxide Dismutase ; metabolism

OBJECTIVESTo explore a sensitive and specific method for detection of bcl-2/IgH gene rearrangement in diffuse large B cell lymphoma (DLBCL), and verify the credibility of the established method.

METHODSbcl-2/IgH hemi-nested PCR primers were designed using the professional primer design software. Fifty-two samples of pathologically diagnosed DLBCL and 10 fresh tonsil tissues were amplified using hemi-nested touch down-PCR to detect bcl-2/IgH gene rearrangement. The PCR products were cloned and sequenced.

RESULTSbcl-2/IgH gene rearrangement was detected in 6 of 52 DLBCL samples and 2 of 10 fresh tonsil tissues using one-way method. By using the hemi-nested PCR for the second round amplification, 5 of DLBCL were positive, but all of the fresh tonsil tissues were negative. The positive PCR products were sequenced and analyzed on the Internet, 3 of 8 cases obtained by one-way method were false positive, 5 positive cases amplified using hemi-nested PCR were all bcl-2/IgH gene rearrangement. PCR products of 3 false positive cases were homologous to BAC331191 and LLNLR-245D11 in human chromosome 19 and RP11-498P10 in chromosome 1.

CONCLUSIONThere are false positive results using common primers for detecting bcl-2/IgH gene rearrangement. The mechanism may be that highly homologous sequences to human genome exist in commonly used primers. The specificity of the diagnosis could be improved by hemi-nested PCR using the combination of primers we designed and the traditional ones.

Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, bcl-2 ; genetics ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the intervention effect of aqueous fractions from Boschniakia rossica (BRAF) on hepatic oxidative stress in mice with liver injury induced by carbon tetrachloride (CCl4).

METHODThe experimental mice were randomly assigned into the normal control group, the model group, the silymarin (positive control) group, as well as high and low dose BRAF groups. Mice were treated intragastrically with silymarin or BRAF once every day for 7 days. At the end of the experiment, CCl4 was injected intraperitoneally into the mice to establish the acute liver injury model. The pathological changes was detected with hematoxylin and eosin (HE) staining, and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), superoxide dismutase (SOD) , catalase (CAT), glutathione peroxidase (GPx), Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, and the contents of reduced glutathione (GSH) and malondialdehyde (MDA) were detected by the colorimetric method.

RESULTBRAF significantly reduced ALT, AST and ALP activities in serum, alleviated hepatic injury induced by CCl4, increased SOD, CAT, GPx and GSH levels in liver, and SOD, Na + -K + -ATPase and Ca2+ -Mg2 + -ATPase activities in liver mitochondria, and decreased the MDA content in liver and liver mitochondria.

CONCLUSIONBRAF reduces hepatic oxidative stress in mice with acute liver injury induced by CCl4, thereby showing the protective effect on mice with acute liver injury induced by CCl4.

Animals ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; enzymology ; metabolism ; pathology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Liver ; drug effects ; enzymology ; metabolism ; pathology ; Male ; Mice ; Mitochondria ; drug effects ; metabolism ; Orobanchaceae ; chemistry ; Oxidative Stress ; drug effects ; Solubility ; Water ; chemistry

OBJECTIVETo evaluate the effect of conservative surgical management on patients with subglottic cancer.

METHODSNine cases with subglottic carcinoma were treated surgically from 1984 to 1999. There were T2N0 lesions in 2 cases, T3N0-1 in 3 cases and T4N0-1 in 4 cases. All the cases underwent partial laryngectomy including partial cricoid resection. Variations of a pedicled thyroid cartilage flap were used for reconstruct the cricoid defect. The pedicle based muscle was thyrohyoid, sterno-thyroid or inferior constrictor. Unilateral neck dissection was performed on 7 cases and bilateral on two.

RESULTSThe function of phonation were preserved in all cases. Eight of nine 8/9 were decanulated. Normal deglutition were achieved for all patients. The 3 and 5 year survival rates were 8/9 and 6/9, respectively.

CONCLUSIONPedicled thyroid cartilage flap is appropriate for reconstruction of the cricoid defect in the conservative surgery of selected subglottic carcinoma.

Adult ; Aged ; Carcinoma, Squamous Cell ; surgery ; Cricoid Cartilage ; surgery ; Female ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Thyroid Cartilage ; transplantation

ObjectiveTo study the quality control methods for 99Tcm-TRODAT-1 kit and injection.MethodsThe appearance,pH,contents of the bases,the labeling yield,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 kits from three different batches were examined.The kit stability was estimated under different conditions.The transparence,pH,radiochemical purity,half-life,asepsis and bacterial endotoxins of 99Tcm-TRODAT-1 injection were tested.ResultsThe 99Tcm-TRODAT-1 kit and injection were both achromous and transparent,with pH values being 5.9 ± 0.1 and 5.5 - 7.0 respectively.The contents of stannous chloride and TRODAT-1 were stable.The labeling yield of the kit and the radiochemical purity of the injection were both ≥95％.The asepsis test demonstrated that the characters of 99Tcm-TRODAT-1 kit and injection were qualified.TRODAT-1 kit was stable at 0 -4 ℃ for 6 months or at room temperature (20 -25 ℃ )for 10 days,and the radiochemical purity of the injection was still ＞ 90％ at room temperature for 8 hours.ConclusionsThe quality control methods for 99Tcm-TRODAT-1 kit are simple and practical.The kit and injection are qualified and can be used for clinical application.

OBJECTIVETo set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.

METHODSFifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.

RESULTSIn the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively. In the exon 48/exon 19/SRY group, the amplification rates of exon 48, exon 19 and SRY in male were 92%, 90% and 94%, the amplification rates of exon 48, exon 19 in female were 94% and 92%, respectively.

CONCLUSIONThe technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene established in this study is highly sensitive, specific and reliable, and is suitable for PGD of deleted DMD with family history.

Dystrophin ; genetics ; Exons ; genetics ; Female ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Preimplantation Diagnosis ; methods ; Reproducibility of Results ; Sequence Deletion ; Sex Determination Processes