BACKGROUND: Cell division cycle 2 (cdc2) plays an important role in the course of neuronal degeneration in neurodegenerative diseases such as Alzheimer's disease. The silencing of cdc2 gene with adeno-associated virus vector might protect the neurons in neurodegenerative diseases.OBJECTIVE: To pack recombinant adeno-associated virus (rAAV) encoding cdc2-siRNA. DESIGN, TIME AND SETTING: Blank control experiment was performed in the laboratory of Department of Neurology at Wuhan Tongji Hospital between October 2007 and August 2008. MATERIALS: Helper Free adeno-associated virus system (pAAV-MCS-EGFP, p-RC, p-Helper) and AAV-293 cells were purchased from Stratagene. METHODS: The pAAV-MCS-U6-cdc2-siRNA-EGFP expressing plasmid was constructed by molecular biological techniques. The cotransfection of this plasmid, together with p-RC and p-Helper into AAV-293 calls was mediated by calcium acid phosphate. The rAAV encoding cdc2-siRNA (rAAV-U6-cdc2-siRNA-EGFP) was packed and harvested. The silencing effect of this virus on cdc2 gene expression in AAV-293 cells was assessed by Western Blot, and its titer was determined by spot hybridization method. MAIN OUTCOME MEASURES: sequencing of pAAV-MCS-U6-cdc2-siRNA-EGFP plasmid inserting U6-cdc2-siRNA; 3 plasmid pAAV-MCS-U6-cdc2-siRNA-EGFP, p-RC, p-Helper cotransfecting AAV-293 cells; cdc2 expression after rAAV-U6-cdc2-siRNA-EGFP infected AAV-293 cells.RESULTS: ①The successful construction of U6-cdc2-siRNA in pAAV-MCS-EGFP plasmid was proved by DNA sequencing. ②Green fluorescent protein was expressed in AAV-293 cells, and the contransfection was working well. ③The cdc2 gene expression in AAV-293 cells was down-regulated markedly after infection of rAAV-U6-cdc2-siRNA-EGFP. ④The titer of rAAV-U6-cdc2-siRNA-EGFP was 1×10<'12> v.g/mL.CONCLUSION: The package of rAAV encoding cdc2-siRNA is successful. It can silence cdc2 gene effectively.

Objective To evaluate the implication of LEEP cervical conization on the outcome of subse-quent pregnancy. Methods The study group comprised 85 women who had a LEEP in Renmin Hoapital of Wuhan University during Jan. 2005 and Jan. 2007 ,and then had a subaequent pregnancy. 109 control women were extracted from outpatient clinic who received antenatal care in the same period with no history of cervical surgery, matching by age, health condition and perinatal stage. The pregnancy outcome of two groups were analyzed. Results Women who had a LEEP were more likely to give preterm delivery than controla (9.88% va 3.70%). But there waa no differ-ence in preterm delivery(χ2=2.97, P>0.05). So were low birth weight infants, preterm premature rupture of mem-branes (pPROM) or cesarean section. On a further study, we found that the time interval between cervical conization and subsequent pregnancy was associated with risk of preterm birth. The shorter time interval, especially shorter than 6 months,the higher risk of preterm birth. Conclusions LEEP cervical conization is not associated with an in-creased risk of preterm delivery, low birth weight infants, pPROM or cesarean section. LEEP conization is a more sol-id choice for women who want to preserve reproductive function. But it would be better for them to have pregnancy plan six months later.

The hospital spirit is not only the spiritual support and vital source for staff, but also the concentrat-ed expression of ideology and faith, value orientation, moral trait, psychology tendencies, culture stereotypes, the unified will and code of conduct for all kinds of personnel.The party organizations are the best helmsmen to culti-vate and carry forward the hospital spirit, which reveals distinctive and profound medical ethical principles and norms.In this paper, taking X hospital as an example, we describe the core function of the party committee′s lead-ership and results in the process of breeding, inheritance and innovation of hospital spirit and reflect ethically so as to provide good references for the current hospital spirit cultivation and promotion.

Objective To investigate the feasibility of C.trachomatis culture in HaCaT human keratinocytes.Methods According to the procedure for C.trachomatis culture in McCoy cells,clinical swab specimens and standard strains of C.trachomatis serotype E were inoculated into HaCaT cells.Iodine staining,a fluorescent monoclonal antibody test and PCR amplification of the endogenous plasmid of C.trachomatis were performed to detect the growth of C.trachomatis in HaCaT cells.Five passages of subculture were carried out for the standard strain of C.trachomatis serotype E in HaCaT cells,and inclusion bodies were counted after each passage.One-factor analysis of variance was conducted by using the software SPSS17 to determine if C.trachomatis was propagated in HaCaT cells.Results Iodine staining showed typical inclusion bodies of C.trachomatis in the cytoplasm of HaCaT cells.Yellow fluorescence-labeled granules were observed in the HaCaT cells under a microscope.Endogenous plasmids of C.rachomatis were successfully amplified by PCR from the infected HaCaT cells.The number of inclusions in HaCaT cells gradually increased at passage 1 through 5.Conclusions C.trachomatis is successfully cultivated in HaCaT cells in vitro,and the standard strain of C.trachomatis serotype E can propagate in HaCaT cells.

AIM:To observe the expression of calcium-sensing receptor(CaSR) of rat cardiomyocytes in anoxia-reoxygenation(A/R) injury and CaSR-induced changes of intracellular calcium;involvement of CaSR in apoptosis and relevant signaling transduction pathway.METHODS:The A/R injury was remodeled in vitro;CaSR,caspase 3 and caspase 9 were deteced by Western blotting.LSCM was used to observe changes of intracellular calcium during reperfusion with or without CaSR agonist.Morphological changes in different groups were observed by light microscopes.Apoptotic cells were measured by TUNEL assay.RESULTS:By LSCM,it was found that the intracellular calcium was significantly increased during reperfusion both in A/R and activator group.Severe injury was found by HE staining in the above two groups,the number of apoptotic cells was significantly increased according to TUNEL assay.The expression of CaSR,caspase 3 and caspase 9 was significantly increased in A/R group and activator group compared with control.CONCLUSION:CaSR is involved in intracellular calcium overload in A/R cardiomyocyte,which can accelerate apoptosis during A/R.

Objective:This thesis aims to construct the evaluation index system ethics management in Beijing medical institutions for application research.Method:The author applied the evaluation system which was agreed and adjusted by expert groups in four medical and health institutions.Result:After three rounds of expert groups have reached the agreement,the Evaluation Index System on Human Research Ethics Management in Beijing Medi-cal Institutions is established,which contains 6 first-class indicators,16 second-class indicators and 39 third-class indicators.The application on four medical and health institutions show that the evaluation index system is practical,operable and is of high degree of distinction.Conclusion:The evaluation index system is scientific,rea-sonable,practical and is of high degree of validity,credibility and distinction.

Objective:To explore the predictive value of vertebral trabecular and endplate HU values on cage subsidence after posterior lumbar interbody fusion (PLIF), hope to provide reference for surgical planning.Methods:All of 72 patients with lumbar disc herniation that underwent PLIF were retrospectively reviewed, who were divided into two groups according to the occurrence of cage subsidence at one-year follow up. Cage subsidence was defined as more than 4 mm subsidence into the vertebrae valuated by CT at one-year follow up. There were 18 patients enrolled into Subsidence group and 54 patients enrolled into N-Subsidence group. The lumbar lordosis, segmental lordosis, intervertebral height, off-bed time, hospital stay, complications, the trabecular and endplate HU values of upper instrumented vertebrae (UIV) and lower instrumented vertebrae (LIV) were compared between the two groups. ROC was used to explore the thresholds of HU values.Results:There were 14 patients presented cage subsidence into the L4, 4 patients presented cage subsidence into the L5. There was no significant difference in lumbar lordosis, segmental lordosis, intervertebral height, off-bed time, hospital stay, or complications between the two groups. Both UIV and LIV trabecular and endplate showed a lower HU value in Subsidence group than those in N-Subsidence group. The most appropriate thresholds of HU value were 146, 172, 307, 254 for trabecular of UIV, trabecular of LIV, lower endplate of UIV, and upper endplate of LIV, respectively.Conclusion:Vertebral trabecular and endplate HU values could effectively predict the cage subsidence after PLIF, patients should be completely informed the risk of cage subsidence and larger cage should be recommended if they presented HU values under the certain threshold.

OBJECTIVETo compare the efficacy of telbivudine monotherapy and telbivudine combination therapy with adefovir in patients with nucleos(t)ide-naive chronic hepatitis B, high-level hepatitis B virus (HBV) load and hepatitis B e antigen (HBeAg)-positivity, and to explore the relationship between treatment regimen adherence and treatment outcomes.

METODSA retrospective study was performed with 123 HBeAg-positive, high-level viral load (HBV DNA≥10(7) copies/ml), nucleos(t)ide-naive chronic hepatitis B patients. Fifty-three of the patients received combination therapy with telbivudine and adefovir dipivoxil,while 70 patients received the telbivudine monotherapy. All patients were tested for rates of conversion to HBV DNA-negative status,alanine aminotransferase (ALT) normalization, HBeAg seroconversion, drug resistance, and side effects at treatment weeks 12, 24, and 48. Treatment regimen adherence was assessed through self-reporting, and interviews were used to explore the relationships to treatment outcomes. The chisquare test, t test and Fisher's exact test were used for statistical analyses.

RESULTSThe rates of HBV DNA negative conversion in the combination group at treatment weeks 12, 24 and 48 were 62.3% (33/53), 88.7% (47/53) and 94.3% (50/53) and were significantly different from those in the monotherapy group at weeks 12 and 24.The rates of ALT normalization were significantly different between the two groups at treatment week 12 (94.3% vs. 77.1%). The rate of HBeAg seroconversion in the combination group at treatment week 48 was 39.6%, and significantly different than that of the monotherapy group. The rates of drug-resistance in the combination and monotherapy groups at treatment week 48 were 3.8% and 11.4%, and the proportion of non-adherence to the treatment regimen was 53.3%, which significantly affected treatment outcome. No side effects occurred in either treatment group.

CONCLUSIONTelbivudine combination treatment with adefovir was more effective than telbivudine monotherapy and elicited a low drug resistance rate in nucleos(t)idenaive chronic hepatitis B patients with high-level HBV load and HBeAg-positivity. Adherence to the therapy regimen was a key factor influencing treatment outcomes.

Adenine ; analogs & derivatives ; Alanine Transaminase ; Drug Therapy, Combination ; Hepatitis B e Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Organophosphonates ; Retrospective Studies ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Viral Load

Through the questionnaire survey on the current situation of human research ethical management in Beijing medical and health institutions, this paper analyzed the problems existing in ethical management of Beijing medical and health institutions and put forward corresponding countermeasures and suggestions. The results showed that overall status of human research ethical management in Beijing medical and health institutions was satisfying. But the ethical management levels of different types of medical and health institutions were quite different, and medical and health institutions, health family planning administration departments and academic teams should all make corresponding contributions.

With the population aging, the prevalence of chronic kidney disease(CKD)is increasing.Frailty is a complex syndrome in the elderly.Elderly CKD patients have higher risks of frailty and cognitive impairment than the general elderly population.In recent years, the relationship between frailty and cognition has gradually attracted the attention of researchers at home and abroad.Cognitive frailty is regarded as a subtype of frailty and has become one of the research hotspots in the field of gerontology.However, there are few studies on the relationship between CKD and cognitive frailty in the elderly.This article reviews research progress on the topic, including the epidemiology, evaluation methods and possible pathogenesis of cognitive frailty in elderly CKD patients.