Objective To investigate the role of the clock gene promoter methylation in aging. Methods C57BL mice of 4- (young, n=9) and 20- (old, n=10) month-old were determined the promoter methylation level of clock genes (Per1/2, Bmal1/2, Cry1/2, Clock, Npas2) in the stomach, spleen, vascular, kidney and striatum with methylation-specific polymerase chain reaction (MSP). Results The incidence of promoter methylation of Cry1, Bmal2 and Npas2 in spleen increased in old mice (P<0.05), while the promoter methylation of Per1 in stomach decreased (P<0.05), and the promoter methylation of Bmal1 in vascular increased (P<0.05). Conclusion Promoter methylation of some clock genes is involved in process of aging in a tissue-specific way.

AIM　To study the electrochemical behavior of ofloxacin at Pt/GC ion implantation modified electrode. METHODS　With Pt/GC ion implantation modified electrode as working electrode, the behavior of ofloxacin was studied by voltammetry in 0.40 mol*L-1 KCl solution. RESULTS　A sensitive reductive peak of ofloxacin was obtained by linear sweep voltammetry. The peak potential was -1.35 V (vs SCE). The peak current was proportional to the concentration of ofloxacin over the range of 1.0×10-6-3.0×10-5 mol*L-1 with the detection limit of 5.0×10-7 mol*L-1. The behavior of reduction wave was studied and applied to determination of ofloxacin in tablets. CONCLUSION　The reduction process was irreversible. The element composition, atomicity form and depth of distribution at the surface of Pt/GC electrode were determined by Auger electron spectroscopy (AES), X-ray photoelectron spectroscopy (XPS) and scannig electron microscope (SEM). The catalysis behavior and reaction mechanism at Pt/GC modified electrode was also studied.

To screen and identify the possible pathogen of the firstly confirmed human case of avian influenza A in Beijing, the throat swabs and tracheal aspirates of this case were collected and the H5N1 viral nucleotide was tested with real time RT-PCR. The certification of result, screening of other pathogens in respiratory tract and sub-typing of influenza viruses were made by using re-sequencing microarray. It was found that the H5N1 viral nucleic acid was positive in the tracheal aspirate of this case by means of detection with real time RT-PCR and the specific sequence of the non-structural protein (NS) gene of H5N1 virus was obtained through the detection with re-sequencing clip. Through the comparative study with the sequence in Genbank, it was proved to be the H5N1 nucleic acid of avian influenza viruses and excluded the possibility of infections with 30 subtypes of influenza viruses and 33 other respiratory tract pathogens. It is apparent that the pathogen detection with re-sequencing clip shows the high sensitivity and specificity and it plays an important role in the pathogen screening and identification for the firstly confirmed human case of avian influenza A in Beijing.

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

Objective To study the changes of hippocampal caspase-3 and PSD-95 expression levels in the mice exposed to ketamine 30 mg/(kg·d)for three months. Methods Forty C57BL/6 mice were randomly divided into two groups,and the chronic ketamine addiction model was established by giving mice a three month course of daily intraperitoneal injections of ketamine. Immunohistochemical study and Western blot-ting were applied to observe the expression of caspase-3 and PSD-95 protein. Results There were more expression of caspase-3 and less of PSD-95 in ketamine group as detected by immuohistochemistry. Western blotting results showed caspase-3 active fragment level significantly increased com-pared to saline group,but PSD-95 protein level was decreased. Conclusion The increased level of caspase-3 protein and reduced expression of PSD-95 are observed after long-term ketamine administration. These findings may provide an evidence for the neurotoxicity in mouse hippocampus of chronic ketamine addition as a recreational drug.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.

Objective To evaluate the safety of the da Vinci Si surgical system in thyroid surgery and to accumulate operation experience .Methods The da Vinci Si surgical system consists of a surgeon's console, a patient-side robotic cart, and high-definition 3D vision system.The robot arm tips were introduced via a single axillary incision or small chest wall ports and attached to the arms of the robot .The surgeon , sitting at the con-sole, manipulated the EndoWrist instruments and endoscope view that transfer the surgeon 's movements to the arm tips.The so called EndoWrist'technology offers seven degrees of movements ( up, down, left, right, twist, et al) , thus exceeding the capacity of a surgeon's hand in open surgery .The da Vinci Si surgical system was used to perform thyroidectomy in two small pigs .The animals were intubated and kept anesthetized with halothane .Re-sults Four surgical procedures were done using the da Vinci Si system from beginning to the end , including 2 thyroidectomies and 2 thymectomies .No conversions to open or laparoscopic procedure was used .The procedure length was 61 minutes and 50 minutes, respectively.Conclusions Robotic thyroidectomy can be performed safe-ly by the experienced surgeons after short-term da Vinci Si surgical system training .The learning curve for robotic thyroidectomy is shorter .

OBJECTIVE:To improve dispensing quality of small package of TCM formula granule, reduce the rate of dispensing error and shorten dispensing time. METHODS:Key influential factors for small package of TCM formula granule dispensing in our hospital were found out from 5 respects of"man,machine,environment,method,material";two cycles of PDCA cycle management intervention were conducted. Dispensing internal error and dispensing time were compared before PDCA cycle management intervention (Jul.-Dec. 2015,group A),first cycle of PDCA cycle management intervention (Jan.-Jun. 2016, group B) and second cycle of PDCA cycle management intervention (Jul.-Dec. 2016,group C),the effects of management were evaluated. RESULTS:By improving dispensing environment,establishing dispensing standard operation procedure,formulating reward and punishment system,staff training,after 2 cycles of PDCA cycle management intervention,the rate of small package of TCM formula granule dispensing error in TCM pharmacy of our hospital decreased from 4.19％(170/4061) before intervention that of group A to 1.69％(85/5043) and 0.98％(53/5408) in group B and C after intervention (P<0.05 or P<0.01). Average dispensing time of each prescription with 7 doses were shortened from(9.08±2.56)min before intervention(group A)to(7.37± 1.98),(5.97±1.64)min in group B and C after intervention(P<0.05). CONCLUSIONS:PDCA cycle management intervention in our hospital can reduce the rate of small package of TCM formula granule dispensing error and shorten dispensing time.

Objective: To investigate the usefulness of loss of CIC expression as the prescreening detection of 1p/19q co-deletion in the diagnosis of oligodendroglial tumors and its prognostic implication. Methods: The retrospective study included 113 oligodendroglial tumors diagnosed in the Department of Pathology, Xuanwu Hospital, Capital Medical University. Expression of CIC protein was detected by immunohistochemistry, and the 1p/19q co-deletion by fluorescence in situ hybridization in all the tumors; and the correlation of the loss of protein and 1p/19q co-deletion with prognosis was assessed. Results: The rate of negative CIC protein expression was 59.3% (67/113) in 113 oligodendroglial tumors. CIC protein expression was differentially lost in various gliomas, 85.7% (42/49) in pure oligodendrogliomas and 39.1% (25/64) in mixed oligodendroglial tumors (P<0.01). The loss of CIC protein expression showed a sensitivity of 76.1% (54/71), specificity 71.1% (27/38), false positive rate of 16.9% (11/65), and a false negative rate of 38.6% (17/44). In 63 cases integrated diagnosis as oligodendroglial tumors with mutant IDH and 1p/19q co-deletion, the loss of CIC protein expression was 81.0% (51/63); the sensitivity and specificity were increased to 81.0% (51/63) and 76.9% (20/26), and the false positive rate and false negative rate decreased to 10.5% (6/57) and 37.5% (12/32), respectively. By using Kaplan-Meier analysis, the CIC negative group showed a trend towards better outcome than the CIC positive group, but there was no statistical difference (overall survival: P=0.218; progression free survival: P=0.249). Conclusions Detection of the lost CIC protein expression can predict the chromosome 1p/19q co-deletion. In oligodendroglial tumors with IDH mutant and 1p/19q co-deletion, there is no relation between prognosis and CIC protein expression.

Objective: To investigate the diagnostic and prognostic implications of ATRX mutation and p53 mutation in patients with glioma. Methods: The clinicopathologic and molecular features of Chinese adult glioma patients, including diffuse and anaplastic astroastrocytoma with IDH mutation, oligodendroglioma and anaplastic oligodendroglioma with IDH mutation and 1p/19q co-deletion and diffuse astroastrocytoma with IDH wild type were reviewed and tested for ATRX loss expression and p53 overexpression. Results: Loss of ATRX expression was seen in 85.19% (23/27) diffuse and anaplastic astroastrocytoma with IDH mutation, higher than that of oligodendroglial tumors (0/53; P<0.01). Loss of ATRX expression was strongly linked to p53 overexpression(69.57%, 16/23). The patients who lost ATRX expression combined with normal p53 expression survived longer(P=0.013). Conclusions ATRX mutation is a molecular marker for astrocytic tumors. ATRX mutation combined with p53 mutation can predict prognosis of patients with glioma.