BACKGROUND: Differentiation of embryonic stem (ES) cells into cardiomyocytes in vitro has been studied in great detail in the world. However, much of what is currently known about cardiomyocyte differentiation from ES cells has been learned from studies on mouse in China, few studies are on human ES cells.OB JECTIVE: To investigate the differentiation effcacy of human ES cells into functional cardiomycytes with the human H14 ES cell line.DESIGN: Suspending method was used to form pseudo-embryo proper of human ES cells so as to observe ratio of pseudo-embryo proper with rhythmic contraction and expression of specific gene of myocardium in various differentiated phases.SETTING: Molecular Biology Laboratory of Stanley Ho Center for Emerging Infectious Diseases, School of Public Health, the Chinese University of Hong Kong.MATERIALS: Human ES cell line H14 was obtained from WiCell Research Institute (Wisconsin, USA) with a license agreement.METHODS: The experiments were carried out in the Molecular Biology Laboratory of Stanley Ho Center for Emerging Infectious Diseases, School of Public Health, the Chinese University of Hong Kong from August to December of 2006.The H14 ES cell colony was used to form embryoid bodies (EBs) by using suspending method. Four days later,pseudo-embryo proper was cultured in gelatin-coating 6-well culture plate (5-10 embryo proper/well) and spontaneously differentiated into moving pseudo-embryo proper. Rhythmic contraction was observed under microscope and RT-PCR was used to detect expression of special genes of myocardium.MAIN OUTCOME MEASURES: Ratio of pseudo-embryo proper with rhythmic contraction and expression of specific gene of myocardium in various differentiated phasesRESULTS: Spontaneously contracting cells appeared as cluster and were identified in approximately 2% of EBs at differentiation day 8 and increased to as many as 10% of the EBs by day 16. The beating rate of contracting cells arranged at 70-100 beats per minute. RT-PCR analyses demonstrated that cells isolated from spontaneous beating areas within the EB expressed the cardiac transcription factors GATA-4 and Nkx2.5, cardiac progenitor gene Isl-1 and cardiomyocyte marker gene α-MHC.CONCLUSION: This is the first time to report human ES cells can effectively differentiate into functional cardiomyocytes in China.

Objective To investigate the expression and clinical significance of miR-125b in patients before and after treatment of acute myeloid leukemia (AML) with different types.Methods The levels of miR-125b were measured by using real-time fluorescence quantification PCR (qRT-PCR) in 65 AML patients (all AML samples from the Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University,and the control samples isolated from cord blood which were obtained after normal full-term delivery of babies) before and after treatment,and then the relationship between the levels of miR-125b and patients＇ sex,age,peripheral blood cells,type clinically,relapse and therapeutic effect were analyzed.Results Of newly diagnosed AML patients and the control samples,miR-125b positive rate was 100％.The miR-125b expression levels in the control group was 1.50,and its expression in AML was 11.06,and the difference was significant (P =0.036 6).In complete remission (CR) group of acute promyelocytic leukemia (M3),the expression of miR-125b after induction therapy was significantly reduced,and CR rate of miR-125b decreased group was 91.3％,while that of the increased group was 50.0％,and the difference was significant (P =0.042 6).The miR-125 b expression level was decreased from 29.7 ± 4.9 to 19.2 ± 6.0 after chemotherapy,and the difference was significant (P ＜ 0.036 6).In non-M3 AML,CR rate of miR-125 b decreased group was 86.7 ％,while that of the increase group was 42.1％,and the difference was significant (P =0.021 5).There was no correlation between miR-125b expression and patients＇ gender,age and peripheral blood cells.Conclusions The differences in expression of miR-125b is very important in disease occurrence and progress.Using qRT-PCR to dynamically detect the expression of miR-125b dynamically in AML patients before and after therapy may predict outcome more precisely and has the potentials as an effective biomarker in determining prognosis,monitoring the risk of recurrence,and guiding the treatment.

Objective To investigate the proliferation,apoptosis and cell cycle and possible mechanisms of different breast cell lines by difluoromethylorithine (DMFO).Methods The growth of breast cancer MDA-435 (ODC GG) cell lines and SK-br3 (ODC AA) cell lines treated with DFMO were observed.The apoptosis and cell cycle were detected by flow cytometry.PCR was applied to detect the changes of A and G alleles of ODC G316A in MCF-7 cells treated with DFMO.Results The growth inhibition rates of MDA-435 and SK-br3 cells treated with 10 mmol/L and 20 mmol/L DFMO after 48 h were 24.1％ and 33.6 ％,46.3 ％ and 53.5 ％,respectively,and there was statistical significance (t =2.134,P =0.021,t =2.213,P =0.019).The growth inhibition rates of MDA-435 and SK-br3 treated with 10 mmol/L and 20 mmol/L DFMO after 72 h were 28.9 ％ and 35.7 ％,54.3 ％ and 65.4 ％,respectively,and there was statistical significance (t =2.434,P =0.015,t =2.489,P =0.013).The apoptosis rates of MDA-435 (ODC GG) and SK-br3 (ODC AA) cells both dealt with 20 mmol/L of DFMO after 24 h,48 h and 72 h were (7.58± 2.06) ％ and (13.88±3.45) ％ (t =2.047,P =0.041),(43.28±14.28) ％ and (59.96±16.42) ％ (t =3.680,P =0.000),(77.87±30.25) ％ and (93.08±32.15) ％ (t =3.293,P =0.000 1),respectively.The proportions of S stage cells MDA-435 (ODC GG) and SK-br3 (ODC AA) cells under the same condition after 24 h,48 h and 72 h were (13.25±2.38) ％ and (12.89±2.21) ％ (P ＞ 0.05),(21.43±3.12) ％ and (12.24±3.55) ％ (t =2.638,P =0.012),(16.32±3.23) ％ and (15.24±3.01) ％ (P ＞ 0.05),respectively.After the treatment by DFMO,the expression of ODC G316A allele A in breast cancer cell line MCF-7 (ODC AG) was reduced (t =3.708,P =0.000),and the expression of G had no significant changes.Conclusion The proliferation inhibition and apoptosis in breast cancer cells treated by DFMO is different in breast cancer cells with different genetic type of ODC G316A.DFMO can inhibit the activity of ODC,and the mechanism may be that DFMO could selectively bind to ODC G316A allele A.

Gastric Lavage ; methods ; Humans ; Mannitol ; administration & dosage ; Norepinephrine ; administration & dosage ; Organophosphate Poisoning ; Pesticides ; poisoning ; Poisoning ; therapy

0.05).Compared with the CSS,GMFM and WV before treatment,there were statistically difference after 6 and 12 weeks treatment in two groups(Pa

OBJECTIVETo investigate the effects of transcutaneous electrical nerve stimulation (TENS) on motor function in children with spastic cerebral palsy (CP).

METHODSAfter signing the informed consent, 78 children with CP, aged 45.6 +/- 8.5 months (36 to 58 months), were randomly divided into a TENS group (n = 40) and a control group (n = 38). All the subjects received standardized functional exercise program. In TENS group, 2 TENS devices were used and the surface electrodes were applied on the spastic musculotendinous and antagonist muscles in the affected lower extremity. TENS lasted for 20 min per session, 5 days weekly for 6 weeks. After 6 weeks, the functional exercise program was applied by the caregivers. Demographic data were recorded, including age, gender, number of the hemiplegic and diplegic CP, level of gross motor function classification system (GMFCS). Clinical assessments included the composite spasticity scale (CSS), D and E dimensions of the Gross Motor Function Measure (GMFM), and walking velocity was determined before treatment and at 6, 12 and 24 weeks after treatment.

RESULTSNo statistically significant differences were found in age, gender, number of the hemiplegic and diplegic CP, level of GMFCS, as well as clinical assessments (CSS, GMFM and walking velocity) before treatment between the 2 groups (P > 0.05). All the children showed a reduction of spasticity (CSS) after 6, 12 and 24 weeks of treatment (P < 0.05). When compared with the results obtained before treatment, the improvement of standing and walking (GMFM), walking velocity was statistically significant after 6, 12 and 24 weeks of treatment (P < 0.05). Furthermore, the differences of CSS, GMFM and walking velocity between the two groups at 6, 12 and 24 weeks examination were also statistically significant (after 24 weeks of treatment: t value was 8.96, 3.14 and 2.35, P < 0.05, respectively).

CONCLUSIONWhen compared with the control group, 6 weeks of TENS treatment on the affected lower extremity was more effective in terms of reducing spasticity and improving functional performance in standing, walking, and walking velocity in ambulant children with the spastic CP.

Cerebral Palsy ; therapy ; Child ; Electric Stimulation Therapy ; methods ; Female ; Gait Disorders, Neurologic ; therapy ; Hemiplegia ; therapy ; Humans ; Male ; Muscle Spasticity ; therapy ; Transcutaneous Electric Nerve Stimulation ; methods ; Treatment Outcome

OBJECTIVETo investigate genotype of wild-type measles viruses circulated in Beijing in 2003.

METHODSThroat swabs specimens were collected from patients seen during an outbreak of measles and from clinically suspected sporadic measles patients in 2003. Vero/SLAM cell lines recommended by WHO were used to isolate measles virus. Four hundreds and fifty nucleotides of COOH-terminal of nucleoprotein (N) genes were amplified by using PR-PCR. The amplified products were sequenced and the sequences were compared with references viruses from GeneBank.

RESULTSEight strains of measles viruses were isolated from throat swabs of patients who came from seven districts and counties of Beijing. Sequence analysis of the 450 nucleotides of COOH-terminal of nucleoprotein (N) genes indicated that these 8 strains belonged to H1a genotype. The average genetic distances of these 8 strains to H1a genotype, Chin9322, H1b genotype, Chin9475 and H1c genotype, Chin9427, were 0.004 - 0.011, 0.026 - 0.031 and 0.015 - 0.022, respectively. The average genetic distances of these 8 strains to H1a genotype, Anhui 01 - 1/Anhui 02 - 2, were 0.000 - 0.009 (0 - 5 nucleotide variation).

CONCLUSIONSMajor genotypes of wild-type measles viruses circulated in Beijing in 2003 were H1a genotype. The genotypes H1c, H1b and H2 may have disappeared in Beijing.

Child ; China ; epidemiology ; Disease Outbreaks ; Genotype ; Humans ; Immunoglobulin M ; blood ; Measles ; epidemiology ; immunology ; virology ; Measles virus ; classification ; genetics ; immunology ; isolation & purification ; Nucleoproteins ; genetics ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; genetics

BACKGROUNDAttention deficit hyperactivity disorder (ADHD) is one of the most common mental disorders during childhood, characterized by the core symptoms of hyperactivity, impulsivity and inattention and puts great burden on children themselves, their families and the society. Osmotic release oral system methylphenidate (OROS-MPH) is a once-daily controlled-release formulation developed to overcome some of the limitations associated with immediate-release methylphenidate (IR-MPH). It has been marketed in China since 2005 but still lacks data from large-sample clinical trials on efficacy and safety profiles. The aim of this study was to evaluate the effectiveness and safety of OROS-MPH in children aged 6 to 16 years with ADHD under naturalistic clinical setting.

METHODSThis 6-week, multi-center, prospective, open-label study enrolled 1447 ADHD children to once-daily OROS-MPH (18 mg, 36 mg or 54 mg) treatment. The effectiveness measures were parent-rated Inattention and Overactivity With Aggression (IOWA) Conners I/O and O/D subscales, physician-rated CGI-I and parent-rated global efficacy assessment scale. Blood pressure, pulse rate measurement, adverse events (AEs) and concomitant medications and treatment review were conducted by the investigator and were served as safety measures.

RESULTSA total of 1447 children with ADHD (mean age (9.52 ± 2.36) years) were enrolled in this trial. Totally 96.8% children received an OROS-MPH modal dose of 18 mg, 3.1% with 36 mg and 0.1% with 54 mg at the endpoint of study. The parent IOWA Conners I/O score at the end of week 2 showed statistically significant (P < 0.001) improvement with OROS-MPH (mean: 6.95 ± 2.71) versus the score at baseline (10.45 ± 2.72). The change in the parent IOWA Conners O/D subscale, CGI-I and parent-rated global efficacy assessment scale also supported the superior efficacy for OROS-MPH treatment. Fewer than half of 1447 patients (511(35.3%)) reported AEs, and the majority of the events reported were mild (68.2%). No serious adverse events were reported during the study.

CONCLUSIONThis open-label, naturalistic study provides further evidence of effectiveness and safety of OROS-MPH in school-aged children under routine practice.

Adolescent ; Attention Deficit Disorder with Hyperactivity ; drug therapy ; Child ; Delayed-Action Preparations ; Female ; Humans ; Male ; Methylphenidate ; administration & dosage ; adverse effects ; therapeutic use ; Prospective Studies ; Treatment Outcome

Objective:To investigate the mechanism of decomposed Zuoguiwan（ZGW） recipes in treating ovariectomized osteoporosis rats. Method:Forty Sprague-Dawley female rats were equally and randomly divided into Sham-operated group, ovariectomized model group, positive group, and low and high-dose ZGW groups. After 12 weeks of administration by gavage, the bone mineral density (BMD) of rats' distal femur was measured by micro-CT, the morphology of bone tissue were observed by hematoxylin-eosin staining (HE), β-cross-linked c-telopeptide of type Ι collagen (β-CTX) and bone-specific alkaline phosphatase (BALP) in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA and protein expressions of β₂AR, OPG and RANKL were evaluated by Western blot analysis and real-time quantitative polymerase chain reaction （PCR）. Result:Compared with Sham-operated group, BMD of rats in ovariectomized model group was decreased (P<0.01), morphology of bone tissue was destroyed, serum BALP was reduced, while β-CTX was boosted (P<0.01)，mRNA and protein expressions of OPG in tibia were reduced, while RANKL were increased, and mRNA and protein expressions of β₂AR in the hypothalamus were decreased (P<0.05, P<0.01). Compared with ovariectomized model group, BMDs of rats in low and high-dose ZGW groups were increased (P<0.01), morphology of bone tissue was repaired, serum BALP and mRNA and protein expressions of OPG in tibia were up-regulated (P<0.05, P<0.01), whereas serum β-CTX and mRNA and protein expressions of β₂AR in the hypothalamus and RANKL in tibia were down-regulated (P<0.05, P<0.01). Conclusion:Yang-nourishing components in decomposed Zuoguiwan recipes can improve BMD of ovariectomized rats by regulating OPG/RANKL pathway mediated by β₂AR. "Seeking Yin in Yang" is a crucial mechanism of Zuoguiwan in treating ovariectomized osteoporosis in rats.