ICH GCP requires that all information of clinical trial should be recorded, processed, and stored in a way that allows the accurate reporting, interpretation and verification. A trial master file (TMF) contains all paper or electronic records/documentations related to a clinical trial. As a tool of the retrospective analysis, the TMF profile should be able to reproduce the full procedure of the trial completely. As a part of TMF profiles, both the accuracy and completeness of clinical data management documentation are important in data integrity. It is helpful to learn the workflow of clinical data management in different stage of a clinical trial, to understand which documents are essential, and why the documentation of clinical data management is important for data integrity. This paper elaborates how to perform the good documentation practice of clinical data management, and suggests that both the precise and efficient document management and regular quality control may ensure the high quality of clinical data documentation management on the basis of an intensive awareness of the overall process of clinical data management.

Objective To develop the cDNA microarray for the combined detection of hepatitis virus, and to study the feasibility of applying the microarray in clinical setting. Methods For the small and simple genome of HBV and HDV, the specific primers of PCR were designed with Primer Premier 5.0 program according to the conserved region of HBV and HDV, and 10 and 4 gene fragments were obtained respectively, which could be used as the probes of gene chip. As for the complex genome of HCV, the technique of restriction display PCR (RD-PCR) was employed. Some of gene fragments were selected which were comparatively more specific and sensitive as microarray probes. In order to explore the experimental conditions of microarray in vitro detection, three types of gene chip were prepared successively including HBV and HDV simultaneous detection, HCV detection and modified HCV detection. Results The hybridized signals on the gene chip showed that the effect in detection was satisfactory. Through the prepared gene chips mentioned above, some probes with good quality were selected and the microarray was prepared for HBV, HCV and HDV simultaneous detection. The diagnostic capability of the microarray was evaluated following the washing and scanning steps. Linearity: Serial dilutions of the target DNA or cDNA showed that a strong linear relationship existed between the various concentrations of target DNA or cDNA and the fluorescence intensities obtained from microarray assay (r=0.990 2, r=0.992 1, r=0.981 9), and that the detection range for the microarray was from 104 to 1011 copies/ml. Specificity: Samples from other viruses such as YFV, JET and DV were also subjected to the test and the results were all negative. Reproducibility: The reproducibility of this assay system was evaluated by repeated measurements, and the within-run coefficient of validation of HBV, HCV and HDV were 7.1%, 7.2% and 6.6%, respectively, while the between-run coefficient of validation was 7.9%, 8.2% and 7.6%, respectively. Accuracy: By using the BLAST and the GenBank database, the identity of the obtained sequences including 16 fragments of PCR, 24 RD fragments of HCV and some positive serum samples were verified, each sequenced product was confirmed to be a genome fragment of expected size. In order to fulfill the request of clinical diagnosis, a modified protocol for microarray detection was established. This new protocol consisted of two hours of hybridization, omitting the steps of prehybridization and purification of samples, and the hybridization temperature was elevated from 42℃ to 52℃, et al. The whole protocol could be completed in less than 8 hours. 98, 42 and 5 serum samples from hepatitis B, C and D patients and 130 samples from healthy people were analyzed, respectively, by microarray assay and real-time PCR (Taqmam method). There was a significant correlation between the results of these assays (HBV, r=0.985 4 and HCV, r=0.958 2, P

Objective To prepare the microarrays for joint detection of HBV and HDV. Methods The specific primers of PCR were designed with Primer Premier 5.0 program according to the conserved regions of HBV and HDV. The PCR fragments were purified and cloned into the pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. The DNA microarray was obtained by spotting PCR products onto the surface of glass slides by robotics. Restriction display PCR (RD-PCR) was used to label the samples. Results After the sequences were aligned, we found that the products of PCR amplification were the specific gene fragments of HBV and HDV. The hybridized signals on gene chips indicated that the specificity and sensitivity of DNA microarray for joint detecting the HBV and HDV were satisfactory. Conclusion Using PCR amplification products to construct gene chips is a quick, simple and effective method for clinical diagnosis of HBV and HDV. Further application of restriction display PCR technique in labeling the sample may expedite and raise the sensitivity in multi-virus detection by microarray technology.

Objective To investigate the clinical effect of surgical treatment for hilar cholangio-carcinoma. Methods The clinical data of 89 patients with hilar cholangiocarcinoma surgically treated in our hospital were retrospectively analyzed. They were divided into 3 groups: radical resection(group A,n=23),palliative resection (group B,n=44) and external drainage operation (group C,n=22). Complications,operative mortality,survival rate and posttreatment quality of lire were compara-tively analyzed among the 3 groups. Results The rate of complications was significantly higher in group A than in group C (P<0.05). There was no marked difference in operative mortality between group A and group B (P>0.05). The 1-,2-and 3-year survival rates and scoring of quality of life were remarkably higher in group A than in other 2 groups (P<0. 001 and 0. 05). Conclusion Radical re-section of hilar cholangiocarcinoma can improve the long-term survival and significantly enhance quality of life of the patients after operation. For patients receiving unradical resection, palliative surgical man-agement can improve the long-term survival and enhance quality of life.

The candidate donor cells for repairing the central nervous system included olfactory ensheathing cells,oligodendrocyte progenitor cells and Schwann cells. Among them,oligodendrocyte progenitor cells are difficult to collect in a large amount; Schwann cells are difficult to traverse glial scar,so olfactory ensheathing cells were the most appropriate groups. Olfactory ensheathing cells in vitro were flexible and of plasticity,thus were capable of adapting to the transplantation microenviroment and benefit for the neural regeneration. Olfactory ensheathing cells could improve the function after injury of spinal dorsal roots,which were probably related to the component of grafts. The proper preparation and mixed olfactory ensheathing cells could contribute to recovery of function. Although the low immunogen of fetal brain,the administration of immunosuppressant would be necessary. In spite of reconstruction of damaged pathway of nervous system,olfactory ensheathing cells were able to promoting sprout of fibers,release neurotransmitters at the non-synaptic sites and improve microenvironment of damaged sites as well,which compensated for the dysfunction in central nervous system injury. Insight into cell biological property and behavior after transplantation would help understand and exert theoretical influence on the repair of spinal cord injury.

OBJECTIVE:To determine the contents of Cu,Zn and Mn in Ejiao,a kind of Chinese traditional medicine.METHOD:The contents of Cu,Zn and Mn were determined by derivative flame atomic absorption spectrometry (DFAAS) after digested by HNO3-HClO4.RESULTS:The contents of Cu,Zn and Mn were 10.48,12.38 and 18.09μg.g-1 respectively.CONCLUSIONS:The DFAAS possesses higher sensitivities,lower detection limits and better precision.

MicroRNAs ( miRNAs) , which are endogenous small noncoding RNAs , mainly regulate the expression of many target genes at the post-transcriptional and ( or ) translational levels.Aberrant expressions of several miRNAs were found in a large variety of neoplasms , including hepatocellular carcinoma ( HCC).Previous studies have shown the existence of a large amount of stable miRNAs in human serum, which have laid the foundation for studying the role of serum miRNAs in the diagnosis and prognosis of HCC.

Objective To investigate the feasibility of preparing HCV-1 diagnostic microarray probes using the technique of cDNA fragments library construction. Methods The full-length cDNAs of HCV of subtypes 1a and 1b were digested with restriction endonuclease Sau3A I, and the resulted fragments were cloned into the pMD18-T vector. Positive clones were isolated and identified by sequencing. Results A total of 57 different fragments were obtained, and sequence analysis showed that all the fragments ranging from 200 to 1000bp were specific gene fragments of HCV genotype 1, which can be efficiently used as probes in microarray prepapration. Conclusion The method of preparing microarray probes by construction of cDNA fragments library was effective, quick and simple.

Objective To investigate the applieation of restriction display (RD) technique in the preparation of HCV probes of clinical genotyping microarray. Methods Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV cDNAs of three distinct subtypes, i.e.1a, 1b and 2a. The resultant restrictive fragments were then ligated with universal adapters. PCR primers were designed to match the universal adapters but with one "nesting" base overhanging at the 3′- end. The PCR reactions were performed by ten pairs of different primer combinations. The differential genes were separated through polyacrylamide gel electrophoresis and silver staining. The second-round PCR was performed using the isolated bands as PCR templates. The purified PCR products were then cloned into T-vectors. The recombinant plasmids were extracted from positive recombinant clones and the target gene fragments were sequenced. Results The target HCV gene fragments ranging from 200 to 900bp were isolated and sequenced, which were correlated precisely with the RFLP (Restriction Fragment Length Polymorphism) prediction. A total of 66 different fragments were obtained, averaging about 22 for each subtypes. These fragments could be further used as probes in HCV microarray preparations. Conclusion RD technique is of great value in obtaining a large number of equal sized gene probes, which provide a swift protocol in generating DNA probes for the preparation of microarrays.

Objective To investigate the efficacy of dexmedetomidine as an adjunct to combined intravenous-inhalational anesthesia for removal of foreign body in the trachea in children.Methods Sixty ASA Ⅰ or Ⅱ children,aged 1-4 yr,weighing 8-23 kg,were randomly divided into 2 groups(n =30 each):combined intravenous-inhalational anesthesia group(group Ⅰ)and dexmedetomidine as an adjunct to combined intravenous-inhalational anesthesia group(group Ⅱ).In group Ⅱ,8％ sevoflurane was inhaled by mask to induce sleep after entering the operating room,and the concentration was reduced to 4％ after the children sere asleep.Anesthesia was maintained with iv infusion of dexmedetomidine 0.5 μg/kg,and then iv injection of propofo1 2 mg/kg,followed by iv infusion of propofol at 6 rag· kg-1· h-1 and remifentanil at 0-15 μg· kg-1 ·min-t.In group Ⅰ,dexmedetomidine was not used and the other procedures were the same as those in group Ⅱ.Sevoflurane inhalation was stopped 2 min later and the rigid bronchoscope was inserted.HR and SpO2 were monitored and recorded before insertion and at 1 and 5 min after insertion.Complications such as respiratory depression,laryngeal edema,and bradycardia were reconded.The amount of propofol and remifentanil consumed,satisfactory level of bronchoscopy,and emergence time were recorded after operation.Results Compared with group Ⅰ,HR was significantly decreased at 1 and 5 min after insertion,SpO2 was significantly increased at 1 min after insertion,the amount of propofol and remifentanil consumed was significantly reduced,the operation time was significantly shortened,the emergence time was significanlly prolonged,the satisfactory level of bronchoscopy was significandy increased,and the incidence of respiratory depression and laryngeal edema was significantly decreased in grmp Ⅱ(P ＜ 0.05).Conclnslon The efficacy of dexmedetomidine as an adjunct to combined intravenous-inhalational anesthesia is better than that of combined intravenous-inhalational anesthesia for removal of foreign body in the trachea in children,with fewer complications.