To explore the molecular mechanism of human papillomavirus subtype 16(HPV-16)E7 oncogene-induced DNA re-replication in response to DNA damage. Flow cytometry was performed to examine the cell cycle changes in RPE1 E7 cells stably expressing HPV-16 E7 and its control cell RPE1 Vector after DNA damage.Immunoblotting assay was used to evaluate the early mitotic inhibitor 1(Emi1)expression in RPE1 E7 and RPE1 Vector cells with or without DNA damage.The changes of the proportion of polyploidy was detected by flow cytometry in DNA-damaged RPE1 E7 cells interfered by Emi1 small interfering RNA. Compared with the control cells,the proportion of polyploids in RPE1 E7 cells was significantly increased in response to DNA damage(=6.397,=0.0031).Emi1 protein expression was significantly increased in DNA damaged RPE1 E7 cells(=8.241,=0.0012).The polyploid ratio of RPE1 E7 cells was significantly reduced after Emi1 was interfered by two independent small interfering RNAs(=2.916,=0.0434;=3.452,=0.0260). In response to DNA damage,Emi1 promoted DNA re-replication caused by HPV-16 E7.
DNA Damage ; DNA Replication ; Human papillomavirus 16 ; Mitosis ; Oncogene Proteins, Viral

Objective To find some risk factors correlated with test anxiety of middle school students,and to find out influencing pathway for test anxiety. Methods 647 middle school students were investigated with Sarason' Test Anxiety Scale (TAS), Achievement Motivation Scale (AMS), Coping Style Scale for School Students ( CSS), Eysenck Personality Questionnaire( EPQ), Egma Minnen av Bardndosnauppforrestran(EMBU) and Family Environment Scale-Chinese Version(FES-CV). Statistics were done with version of SPSS14.0,and data were analyzed by Pearson correlation, multiple linenear stepwise regression and path analysis. Results The rates of test anxiety respectively was mild 25.97% ,moderate 45.65% ,severe 28.38%; there were no significant different between the male and female students anxiety ( 16.71 ± 6.44,17.01 ± 7.02, t = 1. 469, P = 0.334). Test anxiety positively correlated with Achievement motivation, reach motivation of competition, endurance, escape, expos, deny the fantasy,family conflicts,parental punished severely,excessive interference,objective deny,overprotective of father.( r 1-16 :0. 214,0. 135,0. 254,0. 216,0. 308,0.472,0. 492,0. 168,0. 249,0. 537,0. 282,0. 102,0. 238,0. 185,0. 233,0.301,0.273; P 1-16 = 0. 000 ～ 0. 030) , and negatively correlated with Problem-solving, rationalizing interpretation, family cohesion, informative, entertaining, emotional expression, organization, parental warmth and understanding ( r1-9: -0. 121, -0. 134, -0. 178, -0. 215, -0. 221, -0. 101, -0. 298, -0. 136, -0. 168; P 1-9 =0.000 ～0.007). Enter test anxiety regression equation is the reached motivation of competition,emotional expose,organization, psychosis, Neuroticism, parent's warm and understanding , mother's refuse and deny ( t 1-7: 2.496,2.521, -2.687, -2. 150,3.503,2.237,2.259; P1-7 =0.001 ～0.038). Conclusion Test anxiety is commonly find in middle school students. Test anxiety is affected by some paths that are personality,achievement motivation,emotional coping style,family environment and parental education methods.

According to the principle of evidence-based medicine (EBM), Chinese medical literatures based descriptive statistical analysis of common Chinese medical syndrome types were performed. By data extraction, standardization, and frequency calculation of disease names and syndrome types from 286 literatures in line with the inclusion criteria, the frequencies of diseases and syndromes were obtained to analyze common syndrome types in clinical practice, to analyze the distribution features of disease related syndromes and syndrome related diseases, to analyze the distribution of basic Chinese medical syndrome types in clinical common diseases as a whole, thus providing reference for clinical and basic researches.
Evidence-Based Medicine ; Humans ; Medicine, Chinese Traditional

OBJECTIVETo evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide.

METHODSNinety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis.

RESULTSThe levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced.

CONCLUSIONIn the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Animals ; Azoospermia ; chemically induced ; metabolism ; Cyclophosphamide ; toxicity ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Epididymis ; metabolism ; Immunohistochemistry ; Insulin-Like Growth Factor I ; biosynthesis ; Male ; Oligospermia ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism

OBJECTIVETo observe the difference in the clinical efficacy on refractory insomnia between the balance needling therapy and the conventional acupuncture and verify the effectiveness and practicality of the balance needling therapy.

METHODSSixty cases were randomized into a balance needling therapy group, a conventional acupuncture group and a placebo group, 20 cases in each one. In the balance needling therapy group, Shi-mian (point for insomnia) was selected with quick needling manipulation. After the arrival of qi, the technique of lifting and thrusting was applied till the radiating numbness presented in the middle or index finnger, thus the needle was removed. In the conventional acupuncture group, Sishencong (EX-HN 1), Shenmen (HT 7) and Sanyinjiao (SP 6) were punctured with the conventional technique. The treatment was given once every day, 7 treatments made one session, 2 sessions were required totally. In the placebo group, starch capsules were prescribed for oral administration before sleep every night, continuously for 14 days. The efficacy and scores of Pittsburgh sleep quality index (PSQI) beforea and after treatment were observed in three groups.

RESULTSPSQI scores were reduced after treatment in every group (both P < 0.01), the scores in the balance needling therapy group and conventional acupuncture group were reduced apparently as compared with those in the placebo group (both P < 0.01). The total effective rate were 90.0% (18/20), 85.0% (17/20) and 15.0% (3/20) in the balance needling therapy group, conventional acupuncture group and placebo group separately. The efficacy in the balance needling therapy group and the conventional acupuncture group was higher obviously than that in the placebo group (both P < 0.01), and the balance needling therapy group had the same efficacy as conventional acupuncture group (P > 0.05).

CONCLUSIONThe balance needling therapy is safe and effective in the treatment of refractory insomnia and achieves the same efficacy as the conventional acupuncture, which has more simple operation and less pain for the patients and much more considerable practicality in clinic.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Sleep ; Sleep Initiation and Maintenance Disorders ; physiopathology ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effect of Jinguo Weikang Capsule [see text] on the gene expression of H-ras, epidermal growth factor receptor (EGFR), P53 and C-myc of the gastric mucosa in rats with gastric precancerous lesions, and to investigate the action mechanism of JWC on gastric precancerous lesions.

METHODSA rat model with paratypical proliferation of the gastric epithelium mucosa was established by using 60Co irradiation. Rats were divided into the normal group, model group, high-, medium-, low-dose JWC treatment groups, and the vitacoenzyme control group, and were treated for 30 days. The expression of H-ras, EGFR, P53 and C-myc genes of the gastric mucosa was detected by using immunohistochemical methods.

RESULTSThe expression and over-expression rates of H-ras, EGFR, P53 and C-myc gene in the high-and medium-dose JWC treatment groups were significantly lower (P<0.05) as compared with those of the model group.

CONCLUSIONJWC can inhibit the expression of the H-ras, EGFR, P53 and C-myc genes expression of the gastric mucosa in rats, which may be one of mechanisms involved in suppressing or reversing gastric carcinogenesis.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Gastric Mucosa ; drug effects ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; drug effects ; Immunohistochemistry ; Oncogene Proteins ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Epidermal Growth Factor ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; ras Proteins ; metabolism

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.

In order to study function of NAC transcription in development, hormone regulation and the stress response of Salvia miltiorrhiza, the NAC transcription was cloned and analyzed. By retrieving cDNA database of S. miltiorrhiza hairy root one NAC unigene was found, then a full length of cDNA was cloned by designing specific primers and PCR amplifying. Using ORF finder it was found that the cDNA containing a NAC-AB conserved domain in N-terminal, so the cDNA was a NAC transcription factor, named as SmNAC1 (kF006346). Bioinformatics analysis showed that SmNAC1 had an open reading frame (ORF) of 591 bp encoding 196 amino acids. The calculated protein had isoelectric point (pI) of 4.36 with molecular weight about 21.66 kDa. The transcription level of SmNAC1 after dealing with yeast extract (YE) and silver ion (Ag+) in S. miltiorrhiza hairy root was markedly stimulated up regulating. It was 1.4 fold compared with the control after induction 2 h, and maintained 2.0 fold on 4-12 h after induction. SmNAC1 may participate in regulation of stress response of YE + Ag+.
Cloning, Molecular ; Computational Biology ; Phylogeny ; Plant Proteins ; genetics ; physiology ; Plant Roots ; chemistry ; Salvia miltiorrhiza ; chemistry ; genetics ; Trans-Activators ; genetics ; physiology

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 observe the efficacy of intravenous scopolamine in the prevention of postoperative nausea and vomiting (PONV) after cesarean section (CS).

METHODSA total of 260 pregnant women with American Society of Anesthesiologists (ASA) Physical Status Classification class I-II who underwent elective CS under combined spinal-epidural anesthesia (CSEA) were randomly divided into four groups (n = 65): at the end of surgery, 0.3 mg/5 ml scopolamine (scopolamine group), 4 mg/5 ml ondansetron (ondansetron group), 0.3 mg scopolamine plus 4 mg ondansetron per 5 ml (combination group), or 0.9% normal saline 5 ml (control group) were intravenously infused, respectively. The episodes of PONV and adverse effects were observed within 24 hours after operation.

RESULTSThe incidences of PONV within 24 hours after surgery were 87.7%, 89.2%, and 92.3%, respectively, in scopolamine group, ondansetron group, and combination group, which were all significantly higher than that in control group (73.8%) (all P < 0.05). However, the incidences of PONV showed no significant difference among these three groups (P > 0.05). No significant difference in the incidence of adverse effects was observed among the four groups (P > 0.05).

CONCLUSIONIntravenous scopolamine (0.3 mg), with a comparable efficacy as ondansetron 4 mg, can effectively decrease the incidence of PONV after CS.

Administration, Intravenous ; Adult ; Cesarean Section ; Female ; Humans ; Middle Aged ; Ondansetron ; administration & dosage ; therapeutic use ; Postoperative Nausea and Vomiting ; prevention & control ; Scopolamine Hydrobromide ; administration & dosage ; therapeutic use ; Treatment Outcome