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 identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Objective To investigate the prevalence,genotype and epidemiology of plasmid- mediated AmpC enzyme of Escherichia coli and Klebsiella pneumoniae.Methods A total of 67 clinical isolates of nonrepetitive cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae collected by Fuzhou General Hospital and Fujian Provincial Hospital during a period of Sept.2004 to Mar.2005 were detected by three-dimensional extract test for AmpC enzyme,and PCR for AmpC enzyme and other ?-lactamase gene amplification and DNA sequencing were carried out for genotype of ?-lactamase.Plasmid transformation experiment was used to study the transfer of cefoxitin resistance.The homology of the isolates was determined by ERIC-PCR fingerprinting.Results At two hospitals in Fuzhou,the prevalence of plasmid-mediated AmpC enzyme among cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae were 16.7% and 10.5%, 8.0% and 0,respectively.Two isolates of Klebsiella pneumoniae produced DHA-1 plasmid-mediated AmpC enzyme,and 4 isolates of Escherichia cob and one strain of Escherichia coli produced CMY-2 and CMY-22 plasmid-mediated AmpC enzyme respectively.Furthermore,5 strains of Escherichia coli with CMY AmpC enzyme were also found simuhaneously to produce TEM-144,CTX-M-27,CTX-M-14 and TEM-1 ?-lactamase respectively.Three strains of Escherichia coli and one isolate of Klebsiella pneumoniae could transfer cefoxitin resistance to acceptant bacillus.ERIC-PCR fingerprinting reveals 2 strains of Klebsiella pneumoniae came from same clone,but 5 strains of Escherichia coli came from different clones.Conclusions The clinical isolates of Klebsiella pneumoniae producing DHA-1 plasmid-mediated AmpC enzyme and Escherichia coli producing CMY-2,CMY-22 plasmid-mediated AmpC enzyme are found in Fuzhou.CMY-22 AmpC enzyme and TEM-144 ?-lactamase are the first reported in the world,GenBank accession number: DO256079,DO256080

Objective To characterize the audiological features in the infants with otitis media with effusion(OME)and to investigate the utility of variety of objective andiometry methods in diagnosis and intervention on OME.Methods Fifry six infants(40 males and 16 females)were investigated,who were referred to our clinic at the General Hospital of Chinese People's Liberation Army by the other hospitals from December 2004 to June 2007 when the infants were diagnosed or highly suspected of OME.The ages at the initial diagnosis ranged from 42 days to three years,with an average of five months.The infants,after receiving the conventional otolaryngological exams,were subjected to the tests of auditory brainstem response (ABR),otoacoustic emission(OAE),tympanometry(226 Hz and 1000 Hz)and behaviors audiometry.Results Among 56 affected infants.87 ears were diagnosed with OME,of which 31 infants were affected bilateral and 25 with monaural.For the 49 infants who received hearing screening at birth.36 infants were referred at the initial screening.For the 52 infants who received repeated screening.all subjects were referred.Six infants without receiving hearing screening came to clinic when their parents observed their kids'hearing impairment.Among the 52 cases(104 ears)who received tympanometry test,20 subjects (28 ears)showed B or C type tympanometry curve.Thirty-nine cases(78 ears)were given tympanometry test at 1000 Hz,of which 38 cases(55 ears)showed abnormal hearing.Among 56 infants(112 ears)with ABR test,49 subjects(74 ears)exhibited prolonged ABR type Ⅰ curve.All 56 infants(112 ears)received OAE test,of which 55 subjects(81 ears)were referred Four infants(8 ears)accepted the behavior test and all of them showed A-B Gap.Conclusions The combined tympanometry test at both 226 Hz and 1000 Hz,ABR latency or threshold test,infant's behavior test and OAE,used jointly,enable characterizing better OME in infants.thus helping early diagnosis of this hearing disorder.

With the rapid development of medical genetics,online Medelian inheritance in man (OMIM) manifests a more and more important role in medical genetics teaching.Using the educational form combining ‘ classroom teaching,review writing and seminar’,‘ Query and use of OMIM ’was introduced into the education of medical genetics.Reality practice revealed that this educational practice maintained advanced and timely status of knowledge and deeply activated self-studying and independent thinking ability of students.

Objective To analyze the demands of Chongqing municipal medical and health institutions at grass-roots level for the graduates of clinical medicine,pharmacy and medical laboratory science and biotechnology.Methods The Chongqing medical and health institutions at grass-roots level served as the objects and their professionals and administrators were randomly performed the talents demands investigation by adopting the questionnaire investigation as the principal thing assisted by the interview on the spot or informal discussion.Results In the aspect of talents demands,the demands for internist,general practitioner and clinical examination staff were greater,which reached to 36.54％,30.77％and 78.10％respectively;in the aspect of professional knowledge demands,the demands for clinical diagnosis,general practice,clinical medication,clinical laboratory and blood test were greater.Con clusion The school talents cultivation does not understand the true requirements of medical and health institutions at grass-roots level and is inconsistent with the practical demands.

Objective We sought to establish the standard operation procedures in department of cardiology.Methods According to the method of constructing standard operation procedures,the standard operation procedures for the department of cardiology was constructed through induction and consulting literature materials.Results 16 standard operation procedures for the department of cardiology were developed,including 8 SOP of common diseases to rescue,2 SOP of common operation in perioperative period,4 SOP for common instruments,2 SOP for common drugs usage.Conclusions Construction of standard operation procedures in department of cardiology can provide the basis for clinical nursing work,it can also supply methodological reference to build the standard operation procedures in other departments.

To explore the effect of magnitude and duration on the performance of Cumulative Sum (CUSUM),with simulation method used on the subject after the insertion of 11 outbreak events into baseline data with Poisson distribution.Sensitivity fluctuated from 9.1％ to 100.0％ with specificities higher than 98.6％.Sensitivity was significantly correlated with magnitude,and increased along with the increase of magnitude.However,no significant correlation was observed between sensitivity and duration.A magnitude which was at least 2.6 times higher than that of the mean daily baseline could result in the sensitivity of 100.0％.Time-lag would be improved along with the increase of magnitude.Time between onset and detection of an outbreak was no longer than one day when magnitude was more than 1.8 of the mean daily baseline.In summary,the performance of CUSUM was influenced by magnitude,but not by duration.CUSUM had the advantage of good time-lag and high sensitivity when the outbreak magnitude was more than 2.4 time over the baseline data.

Objective To validate the effect of nursing diagnosis, outcomes clacication and interventions clacication(NNN Link)on health education in hospitalized patients with coronary heart disease. Methods Ninety-six patients with coronary heart disease in our hospital during July 2016 to July 2017 were randomly divided into experimental group and control group equally. The control group was given the conventional health education, while the experimental group was given health education by NNN Link at days 1,2,3,7 and one day before discharge.The two groups were compared in terms of nursing outcome scores on admission day,the 5th day and discharge day. Result Day 5 after hospitalization and the day discharged,the scores of knowledge in the experimental group were significantly higher than those of the control group(all P＜0.05). Conclusion The linked nursing diagnosis, nursing outcome and nursing interventions can promote the knowledge of health knowledge and help to improve the quality of nursing service so as to promote research and application of standardized nursing language.

Objective To explore the molecular basis of familial hypercholesteraemia(FH)by analyzing the phenotype and genotype relationship through identify the low density liporotein receptor(LDL-r)gene mutation in a FH kindred.Methods A male patient of 15 years old was selected to examine the electrocardiogram,lipid.Color Doppler was used to examine heart and great vessels.The promoter region and the 18 exons of the LDL-r gene were screened by touch-down polymerase chain reaction(PCR)and DNA sequencing.Results The caro-tid intima-media thickness(IMT)was increased to 0.23 cm,while coronary flow velocity reserve(CFVR)was decreased to 1.57,and mode-rate mitral regurgitation was found in the proband.The genetic alteration G→A change at 1 448 of exon 10 causing premature stop codon(W462X).The same heterozygous nonsense mutation was also found in his father.The mutation had been reported in other Chinese patients.In vitro experiments showed that W462X mutation leads to low LDL binding and internalization ability.Conclusions The homozygous mutation(W462X)in exon 10 of the LDL-r gene were identified in the clinically heterozygous FH proband.The W462X mutation is the underl-ying cause of hypercholesterolaemia and clinical AS manifestations.W462X is recurrent mutation among Chinese FH patients.It might be a hot spot mutation in LDL-r in Chinese FH.J Appl Clin Pediatr,2009,24(1):18-20