Aged ; Diabetes Mellitus, Type 2 ; therapy ; Humans ; Walking

Objective To evaluate the contribution of gyrA and parE detection in Ureaplasma genotyping.Methods Sixty Ureaplasma isolates were selected with the Mycoplasma IST kit.The gyrA and parE were amplified by PCR.The DNA was sequenced and compared with the corresponding sequences in GenBank.Results The nucleotide sequence of gyrA had 100% identity in serovar 1,3,6,14 and 100%identity in serovar 2,4,5,7～13,too.But the sequence had 91%identity between the two groups.The nucleotide sequence of parE had 98%～99% identity in serovar 1,3,6,14.And it had 100% identity in erovar 2,5,7,8,11 and 100% identity in serovar4,12,13.But it had only 90% identity between the two groups.Ureaplasma parvum(Up),Ureaplasma urealyticum(Uu)and Up+Uu infection were found 68.3%(41/60),21.7%(13/60)and 10%(6/60) of clinical specimens,respectively.In Up isolates,serovar 3 was 48.8%(20/41).Conclusion Ureaplasma can be divided into two genotypes(Up and Uu)by gyrA analysis.And Up can be divided into four subtypes which correspond to serovar 1,3,6,14,respectively.Serovar 3 is the main isolate in our research.

OBJECTIVETo explore the better therapy in the treatment of ganglion.

METHODSNinety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups.

RESULTSAll of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01).

CONCLUSIONThe two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Ganglion Cysts ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To find out parental locus of control and mental health affecting students test anxiety. Methods The samples were from 5 middle schools including 1000 students,and their parents. They were investigated with the general life scales, Sarason's test anxiety scale, Symptom checklist 90 (SCL-90)and Parenting Locus of Control Scale(PLOC). Results The ratio of test anxiety: the mild was 28.8%, moderate was 45.0%, severe was 26.2% ,and felt anxiety was 55.8%. Ratio of test anxiety was higher in the boy students (X2=9.284, P =0.010) ,and heavier(F:16.42±6.70; M:15.18 ±6.51, t=2.716, P=0.007). Student test anxiety was significantly positively correlated with their parental mental health (r fatherl～11=0.166～0.272, Pfather l～11= 0.000;r mother 1～11=0.182～0.242, P mother 1～11= 0.000); Student test anxiety positively correlated with the effectiveness cof parents education, father's belief on the fate, eontrol of fathers for their children, locus of control from fathers (r1～5=0.075～0.143; P1-5=0.000～0.030);felt anxiety positively correlated with self-expect ,pressure from their mothers,over take care attitude from their parents,self-pressure from their parents( r1～6=0.068～0.230; P1-6=0.000～0.050) ,and negatively correlated with respect attitude of parents for their child, Marital relations,Communication time between the students and their parents,attitude of mother for learning( r1-7=-0.074～-0.140;P1～7=0.000～0.034). Felting test anxiety was served as a dependent factor, some factors enter the regression equation,and they were somatization of father, psychotic mother, the pressure from parents, mother over expectations, self-expectations ,the child control from student's father by Logistic regression analysis ( OR1～16=0.675～3.029;P1-6=0.000～0.007). Conclusion Students test anxiety is a common problem in male and female students. Mental health and locus of control from students' parents show test anxiety has somatization of father, psychotic mother, the pressure from parents, mother' expectations,self-expectations, the control of father for the child 6 risk factors.

OBJECTIVETo prepare a time-resolved fluoroimmunoassay (TRFIA) kit for clinical detection of IgM antibodies to hepatitis B core antigen (HBc).

METHODSImmunocapture method was used to develop the TRFIA kit for detection of the anti-HBc IgM antibodies, and the precision, cross-reactivity and sensitivity of the kit were tested with the clinical serum samples.

RESULTSThe intra- and inter-assay coefficients of variation of the TRFIA kit were 4.8%-7.2% and 7.5%-8.6%, respectively, and no cross-reactivity with anti-HBs, anti-HBc-IgG or anti-HBe was found. Comparison of the results of the TRFIA kit and enzyme-linked immunosorbent assay (ELISA) demonstrated greater sensitivity of the kit than ELISA in detecting the anti-HBc IgM antibodies in 584 serum samples. According to the detection results in 300 serum samples from healthy donors, the cutoff value of the TRFIA kit was 4.5 times of the fluorescence value of the negative control.

CONCLUSIONThis TRFIA kit for detecting anti-HBc IgM antibodies meets the demand for clinical application and can replace the ELISA kits.

Fluoroimmunoassay ; methods ; Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; immunology ; Humans ; Immunoglobulin M ; blood ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

OBJECTIVETo observe the therapeutic effects of beta-elemene combined DC/Dribble vaccine in treating mice with hepatic cancer, thus exploring their anti-tumor mechanisms.

METHODSDentritic cells were derived from Balb/c mice's spleen and their phenotypes were identified. Using hepatic cancer cell line BNL1MEA.7R.1 (abbreviated as BNL) originated from Balb/c mice as target cell, DC/Dribble vaccine was prepared via raising the antigen representing carrier autophagosomes (DRips in Blebs, DRibbles), which were rich in tumor antigen information. The mice previously immunized were divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The PBS was subcutaneously and intraperitoneally injected to mice in the control group. The beta-elemene was intraperitoneally injected at the daily dose of 50 mg/kg to mice in the beta-elemene group and the combined group for 7 successive days. DC/Dribble vaccine was injected into the lymph node of mice in the vaccine group and the combined group on the 1st day, and DC/Dribble vaccine was subcutaneously injected on the 3rd day and the 5th day. All the mice were sacrificed on the 10th day. Their spleens were obtained sterilely, and the suspension was incubated with or without Dribble. The cells were inoculated for 72 h. The contents of IFN-gamma in the supernatant were measured by ELISA. In addition, the spleen cells obtained from the combined group were incubated with different stimulations for 72 h, which were then divided into the control group, the DRibble group, the DC group, and the DC/Dribble vaccine group. The supernatant of cultured cells were collected and the contents of IFN-gamma were measured by ELISA. The liver tumor-bearing mouse model was established, and then the BNL bearing mice were randomly divided into 4 groups, i.e., the control group, the beta-elemene group, the vaccine group, and the combined group. The treatment ways were the same as the immune ways. The tumor size and the survival period were observed in each group. On the 23rd day the mice were sacrificed. The tumor tissue was stripped and stained by HE staining. The pathomorphological manifestations of the tumor tissue were observed by light microscope.

RESULTSIn vitro detection of mice immunized previously by different ways showed that the secretion of IFN-gamma was significantly higher in the combined group than in the rest groups (P < 0.01). The secretion of IFN-gamma was significantly higher in the beta-elemene group and the vaccine group than in the control group (P < 0.01). The spleen cells could be stimulated to secrete a large amount of IFN-gamma in the vaccine group and the Dribble group (P < 0.01). When the beta-elemene was 10 microg/mL as the stimulating dose, the secretion of IFN-gamma obviously increased (P < 0.01). In vivo observation showed that the growth velocity of tumors in mice of the combined group was slowed down. There was statistical difference in the tumor area or the survival period of mice in the combined group, when compared with the other groups (P < 0.01). In HE staining, the surrounding connective tissues of the tumor were wrapped tightly and compactedly, with infiltration of a large amount of inflammatory cells.

CONCLUSIONSbeta-elemene combined DC/Dribble vaccine could induce specific immune cells to secrete secretory cells, thus exerting its anti-tumor effect. Its immunological effects might be associated with enhancing the DC antigen presenting function.

Animals ; Cancer Vaccines ; immunology ; Cell Line, Tumor ; Dendritic Cells ; drug effects ; immunology ; Female ; Liver Neoplasms ; drug therapy ; immunology ; Mice ; Mice, Inbred BALB C ; Sesquiterpenes ; pharmacology

OBJECTIVETo study the effects of Bushen Zhuyun Decoction (BZD) on the reproductive capacity of mice.

METHODSTotally 60 female Kunming mice were randomly divided into the experimental group and the control group. BZD was perfused by gastric tube to mice in the experimental group, twice daily. Meanwhile, mice in the control group were administered with normal saline by gastrogavage. Mice were sacrificed on the 10th day and the 20th day of medication respectively, 15 mice each time in each group. The serum levels of estradiol and progesterone were detected by radioimmunoassay. The morphological changes of uterus and ovary were observed using HE staining. The expressions of leukaemia inhibitory factor (LIF), epidermal growth factor (EGF), and calcitionin (CT) were determined using immunohistochemical assay.

RESULTSCompared with the control group at the same time point, the weights of uterus and ovary increased, the serum levels of estradiol and progestogen increased, the expressions of LIF, CT, and EGF increased in the experimental group on the 10th day and the 20th day of medication (P <0.05, P <0.01). Better results were shown on the 20th day of medication (P <0.05). Histological results showed increased ovarian follicle numbers, increased endometrial gland numbers, endometrial hyperplasia (stratiform arranged), and increased stromal cells in the experimental group, especially on the 20th day of medication.

CONCLUSIONBZD could improve the reproductive capacity by advancing the development of generative organs, promoting the secretion of estradiol and progestogen and the follicular growth, and increasing the receptivity of endometrium.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Endometrium ; metabolism ; Estradiol ; metabolism ; Female ; Mice ; Mice, Inbred Strains ; Organ Size ; Ovary ; drug effects ; Progesterone ; metabolism ; Uterus ; drug effects

OBJECTIVETo study whether N, N'-di-(m-methylphenyi)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide (ZGDHu-1) inhibits proliferation and induces apoptosis in human lung carcinoma cell line EBC-1 cells and its molecular mechanism.

METHODSDifferent concentrations of ZGDHu-1 and different times of culture were used to treat EBC-1 cells in vitro. The inhibition of proliferation was measured by BrdU-ELISA. Cell apoptosis was detected by Annexin V/PI staining and cellular DNA fragmentation ELISA. Phosphorylated p38MAPK and STAT3 were examined by flow cytometry. The protein expressions of bcl-2, bax, p53, Fas, and caspase-3 were detected by Western blot analysis.

RESULTSZGDHu-1 inhibited EBC-1 cell proliferation within a certain range of treating times and does, with a 24 h IC(50) of (295 ± 25) ng/ml, 48 h of (112 ± 8) ng/ml and 72 h of (23 ± 2) ng/ml. The EBC-1 cell apoptosis was confirmed by Annexin V/PI labeling and cellular DNA fragmentation ELISA in a dose-related manner. When EBC-1 cells were treated with 50, 200, and 500 ng/ml ZGDHu-1 for 48 h, the expression rates of phosphor-p38MAPK protein were 67.4%, 88.2%, 91.1%, respectively, and that of the control was 10.6%. That of STAT3 protein were 56.5%, 43.6% and 34.6%, respectively, and that of the control was 89.1%. The expression of bax, p53 and Fas protein was significantly increased, that of bcl-2 was not changed, and that of caspase-3 was significantly decreased by the ZGDHu-1 treatment.

CONCLUSIONZGDHu-1 can inhibit proliferation and induce apoptosis in EBC-1 cells. The mitochondrial pathway mediated by Fas may be one of its mechanisms. The apoptosis of EBC-1 cells may associate with up-regulation of phosphor-p38MAPK and down-regulation of phosphor-STAT3 in the cells.

Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Fragmentation ; Heterocyclic Compounds, 1-Ring ; administration & dosage ; pharmacology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

BACKGROUND: With the development of 3D printing technology, organ and tissue construction can be achieved by constructing a three-dimensional scaffold that is conducive to cell growth. OBJECTIVE: To solve the scaffold over-accumulation during 3D printing.METHODS: Fluent, a finite element analysis software developed by ANSYS Company in the United States, was used to analyze the extrusion process of print heads and to obtain suitable viscosity and extrusion pressure of materials for the 3D printing of cellulose gel composites. We then compared simulation results with experimental results. RESULTS AND CONCLUSION: The error between simulation results and experimental results was less than 5%. The simulated values at a kinetic viscosity of 45 and a pressure of 0.10-0.12 MPa solved the phenomenon of over-accumulation of cellulose gel composites during the 3D printing process, ensuring enough space for the 3D printed scaffold.

OBJECTIVETo investigate the effects of smoking on sperm apoptosis and semen quality of healthy adult males in the main urban area of Chongqing.

METHODSAccording to the smoking habit, we divided 235 healthy adult males into a non-smoking group (n = 89) and a smoking group (n = 146). Then we detected the routine semen parameters by the computer-assisted semen analysis system and obtained the parameters of sperm apoptosis (the ratios of AN-/PI-, AN+/PI-, AN+/PI+ and AN-/PI+ sperm) by flow cytometry combined with Annexin V-FITC/PI fluorescence staining.

RESULTSThe rate of early apoptotic sperm (AN+/PI-) was higher in the smoking than in the non-smoking group ([8.1 +/- 5.1]% vs [6.8 +/- 3.8]%; P = 0.039), but there were no significant differences between the two groups in the rate of late apoptotic sperm (AN+/PI+) ([5.6 +/- 5.2]% vs [5.5 +/- 5.1]%; P = 0.87), as well as in such routine semen indexes as semen volume, sperm density, sperm motility, sperm vitality and normal sperm morphology (P = 0.30, 0.82, 0.37, 0.81 and 0.84, respectively).

CONCLUSIONThe rate of early apoptotic sperm is higher in smokers than in non-smokers, suggesting that smoking may induce early damage to sperm cells. Compared with routine semen parameters, sperm apoptosis is a more sensitive biomarker to reflect smoking-induced damage to sperm.

Apoptosis ; China ; Humans ; Male ; Semen ; Semen Analysis ; Smoking ; Sperm Count ; Sperm Motility ; Spermatozoa ; cytology