The cancer stem cells (CSCs) from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified. The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension, and mixed homogeneously into 1.2% alginate gel. Single-cell alginate gel was cultured with serum-free DMEM/F12 medium. Epirubicin (0.8 mug/mL) was added to the medium to enrich CSCs. After cultured conventionally for 7 to 10 days, most of cells suspended in alginate gel were killed by epirubicin. But few cells survived and some single-cell cloning spheres formed. Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation. Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo. The results showed that some cells positive for Oct3/4 (TRITC) and Nanog (TRITC) were found in single-cell cloning spheres, and most of positive cells were concentrated in the core of sphere. Cells from spheres could form osteosarcoma in the body of mice. It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes (Oct3/4 and Nanog), resisting anti-cancer drugs, and tumorigenicity in vivo. To sum up, it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

OBJECTIVETo compare the effect of transumbilical single-site single-port with that of transumbilical single-site double-port laparoscopic varicocelectomy in the treatment of varicocele in adolescents.

METHODSWe randomly assigned 80 varicocele patients aged 10 - 16 years to two groups of equal number to receive transumbilical single-site single-port and single-site double-port laparoscopic varicocelectomy, respectively. We compared the operation time, postoperative hospital stay, incisional pain, complications and satisfaction with the abdominal cosmetic outcomes between the two groups.

RESULTSAll the operations were successfully performed. The double-port group showed a significantly higher score on the Visual Analogue Scale than the single-port group (4.8 +/- 1.4 vs 3.6 +/- 1.1, t = -4.986, P < 0.01), but there were no significant differences between the two groups in the operation time ([29.8 +/- 4.2] vs [31.2 +/- 4.6] min, t = 1.383, P = 0.171), postoperative hospital stay ([1.95 +/- 0.7] vs [1.82 +/- 0.8] d, t = -0.784, P = 0.436), complications (0 vs 0) and scores on the satisfaction with abdominal cosmetic outcomes (4.6 +/- 0.6 vs 4.8 +/- 0.5, t = 1.253, P = 0.214). No recurrence, umbilical hernia, hydrocele and orchiatrophy were found in the two groups of patients at 6 months after operation, and no visible scar was observed on the abdominal surface.

CONCLUSIONWith strict surgical indications, single-site single-port and single-site double-port laparoscopic varicocelectomies have similar clinical effects in the treatment of varicocele, which leave no scar on the abdominal surface. Single-site double-port laparoscopy needs no special instruments and therefore is worthier of wide clinical application.

Adolescent ; Child ; Humans ; Laparoscopy ; methods ; Length of Stay ; Male ; Operative Time ; Umbilicus ; surgery ; Varicocele ; surgery

OBJECTIVETo explore the best method for prevention and treatment of thrombosis and its mechanism.

METHODSForty SD rats were randomly divided into a blank group, a model group, an electroacupuncture group and a crude herb moxibustion group. In the electroacupuncture group and the crude herb moxibustion group. "Zusanli" (ST 36), "Sanyinjiao" (SP 6), "Pishu" (BL 20) and "Geshu" (BL 17) were punctured or moxibusted for 2 weeks, then pre-thrombosis model was induced by Adrenalin Hydrochloride and ice water method in the model group, the electroacupuncture group and the crude herb moxibustion group, respectively. Molecular markers in venous blood after the model made in 18 hours were detected.

RESULTSAct: vaty of tissue-type plasminogen activator (t-PA) and content of alpha-granule membrane protein (GMP-140) decreased and content of nitrogen monoxidum (NO) increased after electroacupuncture or crude herb moxibustion. The levels of t-PA and GMP-140 in the model group were higher than those in the electroacupuncture group, the crude herb moxibustion group and the blank group (all P < 0.05), and the content of NO in the model group was lower than those in the electroacupuncture group, the crude herb moxibustion group and the blank group (all P < 0.05), while there was no significant difference in t- PA, GMP-140 and NO among the crude herb moxibustion group, electroacupuncture group and blank group (all P > 0.05).

CONCLUSIONElectroacupuncture and crude herb moxibustion can significantly change the contents of t-PA, GMP-140 and NO and there was no significant difference between the two therapies.

Acupuncture Therapy ; Animals ; Disease Models, Animal ; Female ; Humans ; Male ; Moxibustion ; Nitric Oxide ; blood ; P-Selectin ; blood ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thrombosis ; blood ; pathology ; therapy ; Tissue Plasminogen Activator ; blood

Objective: To explore the cardiac function and outcomes in patients of aortic stenosis (AS) after transcatheter aortic valve replacement (TAVR) within 6 months in order to provide the guidance for clinical treatment. Methods: A total of 49 consecutive severe AS patients with surgical contradiction or STS high risk score and received successful TAVR in our hospital from 2013-12 to 2015-12 were studied. Echocardiography and blood levels of NT-proBNP were examined at pre- and 1 month, 6 months after TAVR. Left ventricular ejection fraction (LVEF), aortic valve mean gradient (MG), peak gradient (PG) and peak velocity (PV) were recorded. Based on pre-operative LVEF, the patients were divided into 2 groups: Cardiac dysfunction group, LVEF<50%,n=15 (30.6%) and Normal cardiac function group, LVEF≥50%, n=34 (69.4%). Post-operative cardiac function and blood levels of NT-proBNP were compared between 2 groups. Results: In all 49 patients, the following parameters were significantly improved within 7 days after TAVR: LVEF (56.0±14.6) % vs (52.5±13.8)%, MG (11±5) mmHg vs (58±18) mmHg, PG (21.7±9.5) mmHg vs (93.0±28.6) mmHg, PV (2.3±0.5) m/s vs (4.8±0.7) m/s, blood NT-proBNP level [1831 (1098-3363)] pg/ml vs [3842 (1763-8664)] pg/ml and aortic valve area (1.57±0.43) cm2 vs (0.58±0.23) cm2 allP<0.05. Within 6 months after TAVR, LVEF was continuously increasing especially in Cardiac dysfunction group; MG, PV and NT-proBNP level were continuously decreasing, NYHA grade was continuously improving, allP<0.05. Conclusion: TAVR was an effective treatment in AS patients with surgical contradiction or STS high risk score; it may continuously improve cardiac function, especially in patients with left heart dysfunction.

BACKGROUNDAdvances in catheter ablation procedures for the treatment of supraventricular arrhythmias have created the need to understand better the morphological and electrophysiological characteristics of the inferior nodal extension (INE) and transitional cellular band (TCB) in the atrioventricular (AV) junctional area.

METHODSFirstly, we observed the histological features of 10 rabbit AV junctional areas by serial sections under light microscopy. Then we recorded the action potentials (APs) of transitional cells (TCs) in the INE, TCBs, AV node, and ordinary right atrial myocytes from the AV junctional area of 30 rabbits using standard intracellular microeletrode techniques.

RESULTSUnder light microscopy, the INE appeared to be mostly composed of transitional cells linking upward to the AV node. Four smaller TCBs originated in the orifice of the coronary sinus, the region between the septal leaflet of the tricuspid valve and the coronary sinus, the inferior wall of the left atrium, and the superior interatrial septum, respectively, all linking to the INE or the AV node. Compared with ordinary atrial myocytes, the AP of the TCs in both the INE and the TCBs had a spontaneous phase 4 depolarization (not present in ordinary atrial myocytes), with a less negative maximum diastolic potential, a smaller amplitude, a slower maximum velocity of AP upstroke, and a longer action potential duration at 50% repolarization (APD50) and at 30% repolarization (APD30). The AP characteristics of these TCs were similar to those of the AV node, except that the velocities of the phase 4 spontaneous depolarization were slower and their action potential durations at 90% repolarization (APD90) were shorter. Moreover, APD50 and APD30 of the TCs of the TCBs were shorter than in the case of TCs of the AV node.

CONCLUSIONSThe TCs of the INE and TCBs are similar to slow response automatic cells. They provide a substrate for slow pathway conduction. In addition, repolarization heterogeneity exists in the AV junctional area.

Action Potentials ; Animals ; Atrioventricular Node ; cytology ; physiology ; Female ; Male ; Rabbits

The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8 μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in alginate gel were killed by epirubicin.But few cells survived and some single-cell cloning spheres formed.Immunofluorescent staining for Oct3/4 and Nanog was implemented to find cells with properties of self-renewal and multi-potential differentiation.Cells from cloning spheres were transplanted into BALB/c mice to detect the tumorigenicity in vivo.The results showed that some cells positive for Oct3/4(TRITC)and Nanog(TRITC)were found in single-cell cloning spheres,and most of positive cells were concentrated in the core of sphere.Cells from spheres could form osteosarcoma in the body of mice.It was concluded that cells from single-cell cloning spheres had the properties of the expression of parts of stem cell genes(Oct3/4 and Nanog),resisting anti-cancer drugs,and tumorigenicity in vivo.To sum up,it is believed that cells obtained from osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs are cancer stem cells.

Abstract Brain Breaks is a physical activity program that combines cultural classroom based physical activity with modern technology while providing children with multi level guidance. As an intervention for intermittent sedentary activities, Brain Breaks can improve students physical activity level, thereby improving their physical fitness and positively affecting their motivation to participate in physical activities and positive learning behaviors. The paper understands this intervention from the connotation, implementation basis, and application effect of Brain Breaks, and then proposes practical application suggestions and future research directions.When Brain Breaks in the cultural classroom is promoted and practiced in China in the future, attention should be paid to the means of implementation by the teachers, the selection of representative target groups, and the precise implementation plan.At the research level, the effects of motor skills, special group interventions, gender differences, environmental changes, and physiological mechanisms of the Brain Breaks are to be explored.

OBJECTIVETo investigate the effects of Apo A1 and B gene polymorphism on avascular necrosis of the femoral head (ANFH) in north Chinese Han population.

METHODSPolymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technique was used in samples of 143 cases with documented ANFH and 92 healthy control matched by age and sex individuals selected from north Chinese Han nationality. The studied loci include promoter region (-75bp) and the intron 1 (+83 bp) of Apo A1 gene, Eco RI,Xba I of Apo B gene; polymerase chain reaction was used to study 3'-VNTR of Apo B gene.

RESULTSAt -75 bp in promoter, the frequency of A/A genotype in ANFH group was significantly higher than that in control group (P < 0.01), while the frequency of G/A genotype in ANFH group was significantly lower than that in control group (P < 0.01). No difference was found in the frequency of genotype at +83bp in intron 1 of Apo A1 gene, Eco RI, Xba I and 3'-VNTR loci of Apo B gene.

CONCLUSIONApolipoprotein A1 gene A/A substitution at position -75 in promotor is associated with ANFH, the mutation may be one of the sensitive genes of ANFH, first reported inside and abroad. But no evident relationship was found between gene polymorphism of +75 bp loci of Apo A1 gene, Eco RI loci of Apo B gene, Xba I loci of Apo B gene or 3'-VNTR of Apo B gene and ANFH.

Adult ; Apolipoprotein A-I ; genetics ; Apolipoproteins B ; genetics ; Case-Control Studies ; China ; Electrophoresis ; Female ; Femur Head Necrosis ; genetics ; Gene Frequency ; Genotype ; Humans ; Male ; Polymorphism, Genetic

OBJECTIVETo optimize the process for preparing genipin with enzymolyzed geniposide by an orthogonal experiment.

METHODThe optimal enzymolysis process was selected by an orthogonal experiment, with the concentration of geniposide as the index as well as enzyme quantity, pH of enzymolysis solution, enzymolysis time and enzymolysis temperature as considerations.

RESULTThe optimal hydrolysis conditions were as follow: rough genipin samples at the concentration of 40 g x L(-1) were selected and shook on a table concentrator at a speed of 100 r x min(-1), added with beta-glucosidase-geniposide 1 : 1 (weight proportion), with pH of enzymolysis solution at 4.5, hydrolyzation temperature at 50 degrees C, the conversion rate of genipin could reach 85.8%.

CONCLUSIONThe process is so stable and feasible that it can provide theoretical basis for the preparation of genipin with enzymolyzed geniposide.

Hydrolysis ; Iridoids ; chemistry ; Technology, Pharmaceutical

AIM: To study the chemical constituents and bioactivity of the seeds of Crataegus pinnatifida. METHODS: The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatography, and preparative HPLC. Their structures were elucidated on the basis of spectroscopic methods. In addition, the cytotoxic activities of compounds 1-4 were investigated on OPM2 and RPMI-8226 cells. RESULTS: Four compounds were obtained and their structures were identified as (7S, 8S)-4-[2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-1-(hydroxymethyl)ethoxy]-3, 5-dimethoxybenzaldehyde (1), (+)-balanophonin (2), erythro-guaiacylglycerol-β-coniferyl aldehyde ether (3), buddlenol A (4). CONCLUSION Compound 1 is a novel norlignan, while compounds 1-4 exhibited marginal inhibition on the proliferation of OPM2 and RPMI-8226 cells.
Cell Line ; Cell Proliferation ; drug effects ; Crataegus ; chemistry ; Humans ; Molecular Structure ; Nerve Tissue Proteins ; chemistry ; isolation & purification ; toxicity ; Plant Extracts ; chemistry ; isolation & purification ; toxicity ; Seeds ; chemistry