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

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

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.

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 study the role of extracellular matrix (ECM) in neural differentiation of mouse embryonic stem cells (ESCs).

METHODSMouse ESCs were incubated in the ESC conditioned medium, and the formation of embryonic bodies (EBs) were induced in bacteriological dishes using high-concentration all-trans retinoic acid (RA). The EBs were seeded on different matrixes (gelatin, fibronectin, and laminin/poly-L-ornithine) to test their impact on neural differentiation of the ESCs using immunofluorescence assay. The effect of laminin/poly-L-ornithine on the growth of neurites was evaluated with fluorescence microscopy.

RESULTSHigh-concentration RA activated and accelerated the differentiation of ESCs toward nestin-positive neural progenitor cells. Fibronectin supplement in the matrix dose-dependently promoted ESC differentiation into neural progenitor cells, while laminin/poly-L-ornithine increased the growth of the neurites and induced the maturation of the differentiated neural cells.

CONCLUSIONECM plays an important role in neural differentiation of mouse ESCs, and application of FN produces the most conspicuous effect during the differentiation of the ESCs into the neural progenitor cells;laminin/poly-L-ornithine is the most effective during their differentiation into neural cells.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Culture Media ; Embryonic Stem Cells ; cytology ; drug effects ; Extracellular Matrix ; physiology ; Fibronectins ; pharmacology ; Laminin ; pharmacology ; Mice ; Neurons ; cytology ; drug effects ; Peptides ; pharmacology ; Tretinoin ; pharmacology

Chorismate mutase catalyzes the conversion of chorismate to prephenate that is the first committed step in the biosynthesis of the aromatic amino acids phenylalanine and tyrosine. A chorismate mutase gene, designated SmCM1, was isolated from Salvia miltiorrhiza by using RT-PCR. The full length of SmCM1 cDNA consists of 948 nucleotides and has an open reading frame of 765 bp. The deduced amino acid sequence of SmCM1 has 255 amino acid residues which forms a 36.0 kD polypeptide with calculated pI of 6.41 as expected. The putative polypeptide contains a CM_2 super family function domain. Blast W results showed that SmCM1 had 70% of the similarity with Petunia x hybrid CM, 72% of the similarity with Arabidopsis thaliana CM, and 64% of similarity with Populus trichocarpa CM. The transcription level of SmCM1 in root, stem and leaf was analysed by realtime quantitative PCR. The results showed the expression level of the SmCM1 in leaf was highest, and lowest in root. Yeast extract and silver ion joint induction could markedly stimulate the increase of mRNA expression of SmCM1 and its upstream 3-deoxy-7- phosphoheptulonate synthase (DAHPS) and chorismate synthase (CS). It was 7.9, 5.5 and 9.8 times of control on 8 h after induction, respectively.
Amino Acid Sequence ; Chorismate Mutase ; chemistry ; genetics ; metabolism ; Cloning, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Tertiary ; Salvia miltiorrhiza ; chemistry ; classification ; enzymology ; genetics

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

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

Objective: To investigate the epidemical features, etiological and clinical characteristics of HFMD in Suzhou city, from 2011 to 2015, providing the scientific supports for HFMD prevention and control. Methods: In each district of Suzhou city, at least five specimens of mild cases were collected per month, while all of the severe cases were sampled. The RNA from each sample was examined using a commercially available real-time PCR kit. Statistical analysis was performed using SPSS version 11.5 software. Results: We retrospectively analyzed HFMD epidemiological data in Suzhou from 2011 to 2015, a total of 4 552 outpatients in Suzhou city were diagnosed with HFMD, including 2 818 positive specimen, the total positive rate was 61.90%, and there was a significant difference in the positive rates between the adjacent years (χ²=186.09, P<0.0001). From 2011 to 2015 in Suzhou, HFMD mainly affected children aged 1 to 5 years old, 66.17% of them were 1to 3 years old. Enterovirus 71(EV71) and coxsackievirus A16(CVA16) were the predominant viral genotypes in Suzhou from 2011 to 2012, and 2014. In 2013, other EVs were dominant, the severe cases mainly correlated with EV71 subtypes, and the proportion of patients with severe disease was significantly decreased in 2013. Conclusions EV71 and CVA16 are still the important pathogens of HFMD, other EVs also occupy a certain proportion, it is the time to select one or two advantage pathogens from the other EVs into the HFMD monitoring.

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.