BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.

Objective To resolve the difficulty in controlling the transfusion speed by medical personnel.Methods By using liquid drop sampling module,pulsing signals of liquid drop were obtained,and then they were enlarged,shaped and the wrong signals from the top and bottom surface of every drop were excluded before the standard triggered signals were sent out to SCM processing circuit,from which the real-time liquid drop speed and the average drop speed in every minute were obtained.Results Anti-interference and portable transfusion-drop counter could show the real-time liquid drop value and the average liquid drop value in every minute.Conclusion Anti-interference and portable transfusion-drop counter is convenient with simple structure and has been used in clinic.

Objective To investigate whether 5 different chemotherapeutic drugs and their combination of either two drugs could further promote the inhibition on the cell growth of HCC cell line (HepG2) in vitro in the hypoxic and hyponutritional culture medium (HHCM) mimicking the different scenarios of transcatheter arterial chemoembolization (TACE).Methods The cells were treated by 5 drugs for 2 h, 4 h,6 h and 24 h, which include epirubicin (EPI), cisplatin (DDP), mitomycin-C (MMC), oxaliplatin (OXA) and 5-fluorouracil (5-FU) in four concentrations of HHCM (5％, 10％, 25％ and 50％) mimicking the scenarios during TACE and the cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.The combinations of dual drugs treated for 24 h were also tested.Results The sensitive drugs with inhibition rates more than 30％ were EPI, MMC and OXA in 4 different concentrations of HHCM.The sensitivity of the drugs treated for 24 h was significantly increased compared with that for 2 h in 5％, 10％ and 25％ HHCM.The dual combinations did not increase the chemosensitivity of HepG2 cells.Conclusions EPI, MMC and OXA exhibited cytotoxic activity against HepG2 cells in various hypoxia and hyponutrition states.Prolonging the exposure time could increase the sensitivity of drug in HHCM, and the combination of dual drugs cannot enhance the cytotoxic effect.

Objective To establish a HPLC method for the assay of psoralen and isopsoralen in different effective extracts of Fructus Psoraleae. Methods HPLC was carried out on the column of Kromasil RP-C18. The mobile phase was methanol -water(65 ∶35). The flow rate was 1.0 mL/min and the UV detection wavelength was 245 nm. Results Good linearity of psoralen was showed within the range of 10.5 ng～525 ng(r= 0.999 3)and isopsoralen within the range of 9 ng～450 ng (r= 0. 999 9). The content of psoralen and isopsoralen differed in different extractions of Fructus Psoraleae. Among them,the extract C (extracted by ethyl acetate ) contained the highest contents of psoralen and isopsoralen,while the contents of psoralen and isopsoralen were very low in the extract D (extracted by n-butyl alcohol) and E (supernatant of water extract). Conclusion The method is simple,accurate and reproducible. The anti-asthma effect and the dose-effect relationship of the different effective extracts of Fructus Psoralea need further pharmacodynamics study.

OBJECTIVE:To observe the clinical efficacy of Hepatocyte growth-promoting factors(PHGF) combined with Wuji baifeng pills for liver cirrhosis. METHODS:A total of 97 patients with liver cirrhosis were randomly assigned to either control group(n=32,conventional therapy) or treatment group(n=65,PHGF plus Wuji baifeng pills in addition to routine treatment).The course of treatment was 3 months for both groups. Clinical data including cardinal symptoms and signs,hepatic function,blood clotting function,hepatic fibrosis parameters,the inner-diameter of the portal vein and splenic vein(PVD,SVD),spleen thickness(SPT) measured by the color Doppler ultrasonography were monitored before and after treatment. RESULTS:After treatment,the symptoms and signs of the treatment group had better improvement than in the control group,with a markedly higher total effective rate than in the control group(84.62% vs.59.38%,P

OBJECTIVE:To observe the efficacy of small dose of uremic clearance granule combined with colon enema on aged patients with chronic renal failure(CRF) . METHODS:60 aged CRF patients were randomly divided into treatment group(n=30) and control group(n=30) . Two groups were given essential treatment. Control group were treated with medicinal carbon additionally. Treatment group were additionally treated with uremic clearance granule(p.o.) combined with colon enema. The course of treatment was 15 days and curative effects were compared between two groups after 30 days of treatment. RESULTS:30 days later clinical symptom of patients in treatment group was obviously improved and the levels of blood urea nitrogen,serum creatinine,blood uric acid and serum potassium were lower than before treatment with significant difference. CONCLUSION:Small dose of uremic clearance granule combined with colon enema show satisfactory effect on CRF in aged patients as well as simple,practical and good compliance. This therapy is worth of spreading and putting into application.

Background Congenital cataract is the major cause of blindness in children with plentiful clinical manifestations and closely linked with genetics. Objective Present study was to investigate the classification of congenital cataract in Yunnan province.Methods The manifestation characteristics of 184 eyes of 116 patients with congenital cataract in Yunnan province were analyzed.The relationship of performance of various types of congenital cataract between phenotype and hereditary feature were explored. Results All of the 116 congenital cataract eyes from 116 patients were divided into ten types based on the clinical appearance,including the complete cataract,nuclear cataract,posterior polar cataract,anterior polar cataract,coralliform cataract,coronary cataract,pulverulent cataracts,lamellar cataract and blue cataract.Bilateral nuclear cataract and unilateral entire cataract is most prevalent in these patients.Blue cataract and coralliform cataract belong to the less types.Forty-four in 116 cases were found to have the hereditary history and determined as autosomal dominant inheritance.Sporadic cases were determined in 72 cases.Conclusions The analysis of phenotype of the hereditary congenital cataract offers some clues to the classification of congenital cataract.

AIM: To study the expression of hepatitis B virus core gene in Pichia pastoris and to obtain high-level expressed recombinant HBcAg with good immunoreactivity and high specificity. METHODS: HBV core gene was amplified by PCR from plasmid pHBV1 which contained HBV whole DNA sequence. The PCR product was cloned into pGEM-T vector by TA cloning strategy. After confirmed by DNA sequence analysis, the gene of interest was inserted into the yeast expression vector pPIC9. The recombinant plasmid pPIC9-cAg was constructed and transformed into GS115 by electroporation. The recombinant yeast GS115 was induced by 0.5% methanol. The expressed product was analysed by SDS-PAGE,Western blot and ELISA. RESULTS: The restriction analysis and DNA sequence analysis proved that HBV core gene had already been cloned to yeast expression plasmid pPIC9. The expressed HBcAg existed in SDS-PAGE. Good immunoreactivity and high specificity of the recombinant HBcAg have been proved by ELISA and Western blot. The titre of the recombinant HBcAg in the cell lysate was 1∶ 12 800 . CONCLUSION: The recombinant plasmid pPIC9-cAg was successfully constructed. The recombinant HBcAg with good immunoreactivity and high specificity was successfully expressed in Pichia pastoris expression system and can be applied to further developing HBcAb immunoassay. [

AIM:MicroRNAs ( miRNAs) were recognized to play significant roles in cardiac hypertrophy .But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy .This study investigates the potential roles of microRNA-1 (miR-1) and microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy .METHODS:An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC).In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte .RESULTS:miR-1 and-16 expression were markedly de-creased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats .Overexpression of miR-1 and -16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes .Expression of cyclins D1, D2 and E1, CDK6 and phosphorylated pRb was increased in hypertrophic myocardium and hypertrophic cardiomyocytes , but could be reversed by enforced expression of miR-1 and -16.CDK6 was validated to be modulated post-transcriptionally by miR-1, and cyclins D1, D2 and E1 were further validated to be modulated post-transcriptionally by miR-16.CONCLUSION: Attenuations of miR-1 and -16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2, E1 and CDK6, and activating cyclin/Rb pathway.

AIM:To investigate the effect of miR-214 on cardiomyocyte hypertrophy and the expression of the potential target genes . METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular car-diomyocytes (NMVCs).Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Wes-tern blotting, respectively.RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-in-duced hypertrophic cardiomyocytes .Dual luciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly in-creased in the hypertrophic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hy-pertrophy-related genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .