OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

OBJECTIVETo establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

METHODSA Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.

RESULTSClustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).

CONCLUSIONThe establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

Alarmins ; Brain ; Cluster Analysis ; Diarrhea ; classification ; diagnosis ; Hot Temperature ; Humans ; Irritable Bowel Syndrome ; classification ; diagnosis ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies ; Surveys and Questionnaires ; Yang Deficiency

Objective To evaluate the significance of CT assessment for extraarticular anatomy in treatment of displaced intraarticular calcaneal fractures.Methods(1)Measurement of normal calcaneum 40 pieces of adult calcaneum specimen were measured,items of measurement included height of culmination of posterior facet and tuberosity,width of posterior edge of sustentaculum and tuberosity.(2)CT measurement of calcaneum.Transverse(axial)and coronal CT scanning were obtained from 20 feet with displaced intraarticular calcaneal and 20 normal feet as control.Following items were measured in CT scanning:the height of culmination of posterior facet and tuberosity,the coronal talocalcaneal angle,in coronal scanning,the width of posterior edge of sustentaculum and tuberosity,the axial calcaneocuboid angle,in axial scanning.Results(1)Measurement of height of calcaneum height of culmination of posterior facet and tuberosity of calcaneal specimen were(43.07?2.85)mm and(44.69?3.67)mm respectively,and these two items from CT scanning of normal feet were(42.84?1.66)mm,(43.40? 3.01)nun,and from CT scanning of feet with calcaneal fractures were(34.76?3.24)mm,(40.41? 3.69)mm.There was a statistically significant different between these two items for normal calcaneal specimen and for CT scanning of feet with ealcaneal fractures(P

Adult ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cerebral Ventricle Neoplasms ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Fourth Ventricle ; Ganglioglioma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Nerve Tissue Proteins ; metabolism ; Oligodendrocyte Transcription Factor 2 ; Rosette Formation ; Synaptophysin ; metabolism ; Young Adult

Objective To explore the changes of tumor necrosis factor-?(TNF-?) in serum and cerebrospinal fluid(CSF) of children with acute lymphoblastic leukemia (ALL) and acute myelogenous leukemia(AML) and its clinical significance.Methods TNF-? in serum and CSF were measured by radioimmunoassay and CSF samples were obtained from 31 cases of childhood acute leukemia before treatment, on complete remission(CR), and continuous CR.Results Serum TNF-? was in ALL and AML before treatment [(24.35?4.84) pmol/L and(28.65?5.12) pmol/ L],which were significantly higher than those of healthy controls[(11.2 8? 1.69) pmol/L, P

Objective To explore the correlation between Alzheimer's disease (AD), vascular dementia (VD) and carotid artery atherosclerosis. Methods This case-control study was conducted among 64 AD and 42 VD in-patients and out-patient admitted in Guangzhou Brain Hospital between July 2005 and July 2007, involving also 69 age- and gender-matched control subjects. Color Doppler ultrasound was performed to examine the artery shape, intima-media thickness (IMT), shape and size of the arteriosclerotic plaques, and diameter of the common carotid artery at 2 cm from the bifurcation and of the distal internal carotid artery 1.0-1.5 cm from the enlargement of the initial segment. Results Compared with those in the control group, the IMT, vascular diameter and arteriosclerotic plaques in the carotid artery were significantly increased in the dementia patients (P<0.05), but no obvious differences were found between the AD and VD patients. The degree of carotid artery atherosclerosis, represented by the number of plaques, was inversely correlated to the Mini-Mental State Examination scores (r=-0.389, P=0.000). Conclusion Carotid artery atherosclerosis is closely associated with the development of AD and VD, and the degree of atherosclerosis is positively correlated to the severity of dementia.

OBJECTIVETo study the effect of internal fixation with absorbable pins on treatment of displaced radial head fractures.

METHODSFrom May 1999 to May 2004, 16 patients with displaced radial head fractures (Mason types II and III) were treated with internal fixation by absorbable pins. The duration of follow-up averaged 22.6 months (12-58 months). The outcome was assessed on the basis of elbow motion, radiographic findings and the functional rating score delineated by Broberg and Morrey.

RESULTSAll fractures healed within 10 months without avascular necrosis of radial head. The mean elbow flexion loss was 15 degrees (0 degrees-35 degrees), and pronation and supination decreased by 10 degrees (0 degrees-30 degrees) on average compared with those of the contralateral elbow. Five patients had an excellent result, 6 a good result, and 3 a fair result according to the criteria of Borberg and Morrey.

CONCLUSIONSInternal fixation with absorbable pins is an effective method in treating displaced radial head fractures. It can maintain the biomechanical stability of forearm, improve the elbow function and avoid second operation.

Adult ; Biomechanical Phenomena ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Prostheses and Implants ; Prosthesis Design ; Radius ; surgery ; Radius Fractures ; physiopathology ; surgery

OBJECTIVETo study the epidemiological characters and risk factors of renal calculi in Shenzhen for future intervention.

METHODSRisk factors of renal calculi were analyzed by factor analysis and linear structural relation model, followed by fitting and evaluating the model.

RESULTSThe prevalence rates of renal calculi were 8.00% and 5.12% in males and females. The results of LISEREL showed that the total effect of age and sex affecting the development of renal calculi was 0.4614, with directly effect 0.3600 and indirect 0.1014. The direct effects of blood uric acid, blood calcium and body mass index, blood cholesterol, blood sugar were 0.3400 and 0.3200 respectively. The indirect effect of education, exercise and dietary habit affected the development of renal calculi through index of biochemistry and obesity, were -0.0416 and 0.1882.

CONCLUSIONSex, age, obesity and high blood cholesterol, high blood sugar, high blood uric acid, high blood calcium were the direct influencing factors to renal calculi. At the same time, education, exercise and dietary habit were also associated with the disease.

Age Factors ; Blood Glucose ; metabolism ; Body Constitution ; physiology ; Calcium ; blood ; China ; epidemiology ; Cholesterol ; blood ; Exercise ; physiology ; Feeding Behavior ; physiology ; Female ; Humans ; Kidney Calculi ; epidemiology ; Linear Models ; Male ; Prevalence ; Risk Factors ; Sex Factors ; Social Class ; Socioeconomic Factors ; Uric Acid ; blood

OBJECTIVEThe aim of this study was to investigate whether the induction of caspase-8 by γ-interferon (IFNγ) renders neuroblastoma (NB) cells sensitive to tumor necrosis factor related apoptosis inducing ligand(TRAIL).

METHODSCaspase-8 mRNA expression was determined by RT-PCR. The effects of IFNγ, TRAIL, IFNγ +TRAIL and caspase-8 inhibitor+ TRAIL on the growth and apoptosis of NB cells were detected with the methods of reduction rate of Alamar Blue assay and flow cytometry. The relative caspase-8 activity was measured with colorimetric assay.

RESULTSCaspase-8 expression was detectable in CHP212 cells which were sensitive to TRAIL, with an increased expression after treatment with IFNγ. Caspase-8 was undetectable in SH-SY5Y(SY5Y) cells which were resistant to TRAIL, but an increased expression of caspase-8 mRNA was found after treatment with IFNγ. Moreover, TRAIL combined with IFNγ induced apoptosis in SY5Y cells. The relative caspase-8 activity of CHP212 cells increased with the prolonged TRAIL action time. The relative caspase-8 activity of SY5Y cells in the IFNγ+TRAIL group was significantly higher than those of the control, IFNγ, TRAIL and inhibitor groups.

CONCLUSIONSNB cells expressing caspase-8 are sensitive to TRAIL. TRAIL induces apoptosis in NB cells with an increase of relative caspase-8 activity.

Apoptosis ; drug effects ; Caspase 8 ; physiology ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Interferon-gamma ; pharmacology ; Neuroblastoma ; drug therapy ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology