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 explore the current state-trait anxiety among nurses in children's hospital and supply some scientific references for the decision-making to healthy guidance and occupation behavior for the practioners. Methods Investigations were carried out in 410 female nurses in Guangzhou children's hospital by usage of the state-trate anxiety inventory(STAI).The results were analyzed statistically. Results The STAI of nurses in the children's hospital was higher than that of the norreal control (P ＜ 0.01 ).But no difference existed in the aspect of special anxiety (P ＞ 0.05) between them.The special anxiety of female cadre and technical staff was higher than the nurses in the children's hospital (P ＜ 0.01). Conclusion The high level of state-trait anxiety hinted us that we should pay attention to the anxiety status of nurses in the children's hospitals.

Objective To study the mechanism by which a high-intensity pulsed electromagnetic field (HIPEMF) (0.1 Hz,4 T,8 pulses) facilitates the proliferation of neural stem cells by detecting the expression of β-catenin genes and protein.Methods Neural stem cells (NSCs) were isolated from the sub-ventricular zone (SVZ) of neonatal rats and cultured in supplemented,serum-free medium for two weeks.The NSCs were then divided into an experimental group exposed to a HIPEMF for 8 pulses and a control group given sham stimulation.The gene and prorein expression of β-catenin in the NSCs were assayed by RT-PCR and Western blotting on the 1st,3rd,5th and 7th day after the stimulation.Results The NSCs' cloned spheres were round and translucent,and showed red fluorescence after staining with anti-nestin (cy3).The RT-PCR results showed β-catenin genes were highly expressed in the exposed group (significantly more than in the controls).The Western blotting showed that expression of β-catenin protein was also higher in the experimental group,especially at the 7th day after stimulation,a difference which was also statistically significant Conclusion HIPEMF at 0.1 Hz,4 T,in 8 pulse trains can promote NSC proliferation,perhaps through the Wnt/β-catenin signaling pathways.

BACKGROUND: Prostate cancer stem cell is an important reason for the invasion and recurrence of prostatic carcinoma. However, separation efficiency of prostate cancer stem cells is very low.OBJECTIVE: To explore the efficient method for isolating and identifying the prostate cancer stem cells from human prostatic carcinoma cell lines PC-3 and LNCap. METHODS: Prostate cancer cell lines PC-3 and LNCap were cultured in serum free medium (SFM) and serum supplemented medium (SSM), respectively. The percentage of prostate cancer stem cells from different medium was detected by flow cytometry through markers CD133 and CD44, and the properties of prostate cancer stem cells were preliminarily identified using inducing differentiation experiments.RESULTS AND CONCLUSION: PC-3 and LNCap formed sphere cells in SFM, which can be induced into adherent cells after culture in SSM. Higher percentage of CD44+/CD133+cells was obtained from LNCap cells (1.71%; 0.73%) than PC-3 cells (0.59%; 0.32%) in both SFM and SSM. The number of CD44+/CD133+ LNCap cells was more than PC-3 using both methods (P < 0.05), but the efficiency of SFM and SSM had no statistical significance (P > 0.05). However, the culture cycle was longer and number of obtained cells was less by SFM culture, directly influencing functional determination of prostate cancer stem cells. Compared with suspension culture method with SFM, SSM is more convenient and effective in isolating prostate cancer stem cells from LNCap cells.

Objective To Investigate the influence of high-intensity pulsed electromagnetic field (HIPEMF) on neural differentiation of neonatal rats neural stem cells in vitro.Methods Neural stem cells (NSCs) were isolated from the subventricular zone of 3-day-old neonatal rats and cultured with serum-free condition medium for 14 days.All the NSCs were then randomly classified into an experimental group which received the stimulation of HIPEMF (0.1 Hz,4 T,8 pulses) and a control group which received no special intervention.Differentiation of the culture was induced by addition of 10％ fetal bovine serum on the first day after intervention,the morphological changes of cells were observed under the microscope at different time points.The NSCs adhered to the wall and differentiated for seven days,the immunofluorescence was employed to observed and calculate the ratio of differentiated cells with astrocyte marker GFAP or neuronal markers TUJ1.RT-PCR and western blotting were used to measure the expression levels of the differentiated cells based on gene and protein levels,respectively.Results Immunofluorescence staining showed the number of TUJI positive cells in the experimental group(33.4％ ± 5.1％)was significantly more than the control group (26.5％ ± 7.0％),while the number of GFAP positive cells was decreased(23.9％ ± 5.0％) as compared with the control group(36.2％ ± 2.2％).RT-PCR showed that the TUJ1 mRNA expression levels in the experimental group was (1.682 ± 0.086) times of the control group.Western blot showed that the expression of TUJ1 (0.729 ±0.061) in the experimental group was higher than in the control group (0.590 ± 0.157),while the expression of GFAP in the experimental group (0.566 ± 0.056) was less than in the control group(1.034 ± 0.051).Conclusions HIPEMF facilitates differentiation of neural stem cells to neurons,at the cost of reducing astrocytic differentiation.

AIM: To investigate the association between HBV infection and HLA-DPB1 gene in population of Guangzhou Chinese. METHODS: 58 unrelated patients (test positive of HbsAg,HBeAg,HbcAb) and 75 unrelated healthy control individuals were typed by sequencing based typing (SBT) method in their HLA-DPB1 gene. RESULTS: The phenotype frequencies of HLA-DPB1 alleles of patients and control have no significant difference. CONCLUSION: These results indicate that there is no association between HLA-DPB1 gene and HBV infection.

Objective:To explore the role of matrix metalloproteinase 12 (MMP12) in airway macrophages and pulmonary neu-rogenic substance P ( SP ) in the pathogenesis of asthma by analyzing their relationship in different categories of asthmatic patients.Methods:Twenty patients of asthma remission phase ( remission asthma group ) , twenty ones of mild acute exacerbation asthma (mild asthma group) and twenty healthy adults (normal control group) were included,respectively.After lung function was measured,the numbers of macrophage in induced sputum were counted.The expression levels of MMP12 mRNA and protein in sputum macrophages were detected by quantitative reverse transcription polymerase chain reaction and Western blot.The concentration of sputum SP was assayed by enzyme immunometric assay.Results: ( 1 ) Compared with the subjects in normal control group, forced expiratory volume in 1 second%predicted ( FEV1 ) and forced expiratory flow rates at 50% of the forced vital capacity % predicted ( FEF50 ) were much lower and the numbers of sputum macrophages were much higher in the patients in different asthmatic groups.Compared with the patients in remission asthma group,FEV1 and FEF50 were much lower in the ones in mild asthma group.(2) MMP12 expressions in the macrophages and the concentrations of SP in sputum were significantly increased in the patients in different asthmatic groups compared with those in normal control group;Furthermore,MMP12 and SP in mild asthma group were much higher than in remission asthma.(3) In all patients from different asthmatic groups,mRNA expressions of MMP12 in the macrophages were positively correlated with the levels of sputum SP or the numbers of sputum macrophages,whereas negative correlations between mRNA expressions of MMP 12 and FEV 1 or FEF50 were observed.Conclusion: The regulatory imbalance of macrophages′MMP12 and pulmonary neurogic SP may participate in the pathogenesis of asthma and become the potential targets for asthma therapy.

Objective In order to improve cirrhotic liver management,each aspect of the liver's complex blood flow must be understood.This study investigates the protective effect of portal vein occlusion,with hepatic artery preservation,on cirrhotic liver after ischemia and reperfusion.Methods Carbon tetrachlorideand induced cirrhotic rats and normal rats were randomly assigned into 4 groups:normal sham operation (N-SO),cirrotic sham operation (C-SO),portal triad clamping (PTC),and portal vein clamping without hepatic artery inflow control (PVC).During the occlusion,the total 3-minute blood loss from the liver surface cut was weighed.At 1,6,and 24 hours post reperfusion,the serum alapine amino transferas (ALT),the adenosine triphosphate (ATP) of liver tissue,the malonolialdehgde (MDA) of liver tissue,and the morphological changes were evaluated.Result The amount of hemorrhage between the groups ranked as follows:PTC ＜ PVC ＜ N-SO ＜ C-SO (P＜0.05).At 1,6,and 24 hours post reperfusion.the ALT and MDA levels of the groups ranked as follows:PTC ＞ PVC ＞ C-SO ＞ N-SO (P＜0.05).Additionally,each group's ATP level ranked as follows:PTC ＜ PVC ＜ C-SO ＜ N-SO (P＜0.05).With histopathological examination,the hepatic injuries of the PTC and PVC group were more severe than those of the C-SO group,especially in the PTC group.Conclusion Therefore,the technique of portal vein clamping and hepatic artery inflow control can reduce the ischemic reperfusion injury of the cirrhotic rats' liver.

Humans ; Receptor, ErbB-2 ; Staining and Labeling

OBJECTIVETo evaluate the diagnostic value of HGAL and LMO2 expression and compare with CD10 and bcl-6 in follicular lymphoma (FL).

METHODS63 cases of FL were collected from Guangdong General Hospital. The expression of HGAL, LMO2, CD10 and bcl-6 was assessed by immunohistochemistry.

RESULTSThe expression rates of HGAL, LMO2, CD10 and bcl-6 were 98.4% (62/63), 82.5% (52/63), 82.5% (52/63) and 87.3% (55/63), respectively. The expression rate of HGAL was higher than those of LMO2, CD10 and bcl-6, but the differences were not significant (P>0.05). There was no significant difference in HGAL, LMO2 and bcl-6 expression among FL1, FL2 and FL3 cases. The CD10 expression rate of FL1-3A cases was significantly higher than that of FL3B cases(P<0.01).

CONCLUSIONSHGAL and LMO2, especially HGAL, can be used in FL particularly high grade FL as useful germinal center marker.

Adaptor Proteins, Signal Transducing ; metabolism ; Biomarkers, Tumor ; metabolism ; Germinal Center ; Humans ; Immunohistochemistry ; LIM Domain Proteins ; metabolism ; Lymphoma, Follicular ; metabolism ; Neoplasm Proteins ; metabolism ; Neprilysin ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; metabolism