Objective To sequence and analyze the complete genome of two human coronavirus NL63 (HCoV-NL63) strains collected from Beijing Children Hospital .Methods Eighteen pairs of primers were designed according to the gene sequences of HCoV-NL63 reference strain ( HCoV-NL63_Amsterdam 1) and used to amplify the target fragments covering the complete genome of HCoV-NL63 strains.Rapid ampli-fication of cDNA ends ( RACE) and RT-PCR assays were used to amplify the full length genome of HCoV-NL63 strains.Phylogenetic analysis was conducted by using Mega 5.0 software.Results The complete ge-nome sequences of the two HCoV-NL63 strains were 27 538 bp in length, showing a homology of 99.1%in nucleotide sequences .There were 15 consecutive bases deleted from 1a region.The systematic phylogenetic analysis demonstrated that four genotypes of NL 63 virus including A , B, C and D have been identified , and two domestic strains were belonged to the new genotype D .Conclusion The complete genome sequences of two domestic HCoV-NL63 isolates were identified for the first time .This study provided evidence for further investigation on molecular epidemiology of HCoV-NL63 in China .

Objective To investigate the value of fluid resuscitation strategy in septic shock patients by pulse indicator continuous cardiac out ( PiCCO ) .Methods 42 septic shock patients were divided into the PiCCO group(n=26) and the control group(n=16).All patients measured heart rate(HR),mean artery pressure(MAP), central venous pressure(CVP);CI,GEDVI,SVRI,EVLWI,CVP as indicator of fluid resuscitation after 0h,6h,24h of the diagnosis were measured respectively in PiCCO group;CVP as guiding volume resuscitation was measured in the control group .The effect of fluid resuscitation was compared between two groups .To analyse the relationship between CVP,GEDVI and CI in PiCCO group .according to CVP increase 2mmHg ,GEDVI whether elevated 10%.Results After 6h EGDT treatment bundle HR ,MAP,APACHEII score and clearance rate of lactic in PiCCO group improved more than those in control group [(101.3 ±7.8) and (119.4 ±7.2),t=-7.520,P<0.05;(71.8 ±7.6) and (51.5 ±8.9),t=7.873,P<0.05;(17.0 ±3.4) and (22.7 ±4.1),t=-4.978,P>0.05;(53.6 ±11.5) and (-16.5 ±5.2),t =9.283,P <0.05].There were no differences in 28-Day mortality between two groups (t =-2.162,P>0.05),but ICU hospitalization time decreased in PiCCO group [(13.8 ±2.6) and (23.3 ±2.2),t=-5.075,P<0.05].Changes in GEDVI was positively correlated with Changes in CI (r=0.799,P<0.05),while changes in CVP was poorly correlated with CI (r=-0.446,P>0.05).Conclusion Hemodynamic monitoring by PiCCO directed fluid resuscitation strategy can elevate reversal rate .Compared with pressure index CVP ,GEDVI is a sensitive indicator of cardiac preload .Correlation between CVP and GEDVI can reflect cardiac function ,Especially for septic shock patients with cardiac depression .

In this study, authors observed and compared the clearances of different molecule weight substances between different dialysis membranes and different hemopurification methods. The results showed that the clearances of BUN by hemodialysis (HD) with cuprophan membrane, hemodiafiltration (HDF) with polysulfone and high flux dialysis (HFD) with polysulfone were all satisfactory. TACurea were all below 50 mg. Hemodialysis with cuprophan could clean out some middle molecule substances (MMS), but not ?2-microglobulin (?2m). Hemodiafiltration with polysulfone had the best clearances of MMS and ?2m. Hemofiltration (HF) with polysulfone had fair clearances of MMS and ?2m, but the clearances of BUN and Cr by HF were not so good. The clearances of MMS and ?2m by high flux dialysis were lower than those of HDF and HF. The clearances of MMS and ?2m by continuous ambulatory peritoneal dialysis (CAPD) at a single time were lower than those of HF, HDF and HFD, but the general clearances of MMS and ?2m by CAPD per week were two to four times higher than those by hemodialysis with cuprophan. CAPD is a choice method for those hospitals where HF or HDF can not be performed.

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.
Animals ; Cations ; Cell Line ; DNA ; Genetic Vectors ; Kidney ; cytology ; Liposomes ; Magnetics ; Nanoparticles ; Swine ; Transfection

OBJECTIVEWe detected and analyzed the clinical values of total protein (TP), albumin (ALB), globulin (GLB), and ALB/GLB ratio (A/G) of whole unstimulated saliva of healthy people to determine the time of day when saliva composition is relatively stable. We compared the protein concentration and A/G of whole unstimulated saliva of patients with chronic periodontitis with those of healthy volunteers to provide references for diagnostic methods and clinical applications of saliva.

METHODSThe whole saliva of 37 healthy subjects were collected at 8:00, 9:30, 11:30, 13:00, 16:30, and 21:00. Meanwhile, the whole saliva of 24 patients with periodontitis was collected in the morning. Bicinchoninic acid method was used to detect the TP content. Saliva ALB was detected by GF-D800 semi-automatic biochemical analyzer, and the GLB and A/G were calculated. Finally, the results were statistically analyzed using SPSS 19.0.

RESULTSSalivary protein compositions were stable in the morning on an empty stomach. Healthy people: TP, (1 354.35±389.52) µg.mL-1; ALB, (139.55±27.19) µg.mL-1; GLB, (1 211.80±360.73) µg.mL-1; A/G, 0.126 3±0.041 7. Subjects with chronic periodontitis: TP, (2 611.56±231.62) µg.mL-1; ALB, (296.27±17.34) µg.mL-1; GLB, (2 315.69±221.67) µg.mL-1; A/G, 0.156 2±0.017 3. The contents of TP, ALB, and GLB in healthy individuals at different periods within a day showed significant differences (P<0.05), which were mainly reflected in the levels before and after meals. No significant difference was detected in A/G. The concentrations of TP, ALB, and GLB were significantly increased in patients with chronic periodontitis compared with those in healthy volunteers. However, no significant difference existed in A/G.

CONCLUSIONSalivary protein compositions are more stable in the morning than in other periods. Thus, mornings can be set as the time of specimen collection in future research. The concentrations of TP, ALB, and GLB in patients with chronic periodontitis are higher than those in healthy people.

Objective To explore the value of endoscopic ultrasound (EUS) in differential diagnosis of ectopic pancreas and stromal tumor in stomach.Methods The clinical data and endoscopic ultrasound findings of 40 patients with ectopic pancrea and 141 patients with stromal tumor were retrospectively analyzed,including the location,size,growth pattern,layer of origin,internal echo pattern and so on.All patients were diagnosed pathologically.Results Ectopic pancreas were predominantly located in the antrum (92.5％) and stromal tumor were mainly located in the gastric body (45.4％),the antrum (23.4％) and the fundus (22.0％),there was a significant difference in lesion' s location between two diseases (P ＜0.001).There was no statistical difference in the ratio of longest/shortest diameter of the lesion size(P =0.057).But there was statistical difference in lesion size between ectopic pancrease and stromal tumor [(19.98 ± 12.80)mm vs (11.25 ± 3.61)mm].Mural growth pattern was most common in ectopic pancrease (82.5％) and mural growth pattern (37.6％),extraluminal growth pattern (24.8％) and intraluminal growth pattern (22.7％) in stromal tumor (P ＜0.001).The third layer (75.0％) was predominant in ectopic pancrease while the fourth (54.6％) and the second layers (34.0％) in stromal tumor (P ＜0.001).With regard to internal echo pattern,hypoechoic (32.5％),intermediate (27.5％) and heterogeneous (27.5％) was predominant in ectopic pancrease and homogeneous hypoechoic lesion (72.3％) in the stromal tumor (P ＜ 0.001).Conclusions There were statistical differences between ectopic pancrease and stromal tumor in the location,size,layer of origin,growth pattern and internal echo pattern under EUS.EUS plays an important role in identification of ectopic pancrease and stromal tumor.

Objective:To investigate the changes of the vertical height and width of the alveolar bone six months after the alveolar ridge preservation in periodontal compromised molar sites of severe alveolar bone defects with clinical direct measurement,parallel periapical radiographs,and cone-beam computed tomography (CBCT),and to analyze the effect of the three different methods of measurement.Methods:In this study,20 subjects requiring tooth extraction on account of periodontal disease with a total of 23 ex-tracted molars were enrolled.Extractions were performed atraumatically and patients were received alveo-lar ridge preservation procedure with Bio-Ossand Bio-Gide.Clinical direct measurements were taken after tooth extraction and during the implant surgery 6 months later,CBCT scans and parallel periapical radiographs were taken immediately after ridge preservation and 6 months later.The changes of alveolar ridge width and vertical height after six months were measured and analyzed through the above-mentioned three methods and the similarities and differences of the measured effect were compared.Results:There were no significant difference of alveolar vertical height in the center of the extraction sites,the center of distal aspect,and distobuccal aspect between the clinical direct measurements and the CBCT measure-ments (P>0.05),alveolar vertical height in other points and alveolar width measurements were statical-ly significant (P<0.05).After 6 months,1 0 sites of 1 0 subjects were received a flap and re-entered to perform dental implants surgery.The vertical height in the center of alveolar increased significantly and the changes of alveolar vertical height of clinical direct and CBCT measurement were (6.1 5 ±1 .73)mm and (6.59 ±2.53)mm,respectively.The measurements of the width of the alveolar bone were (8.45 ± 1 .1 8)mm and (8.52 ±1 .27)mm,respectively.The measurements of the two methods were not statisti-
cally significant (P>0.05).The change of the alveolar height in the center of the extraction socket after six months measured by parallel periapical was (5.84 ±4.28)mm,which was closed to the clinical di-rect measurement and the CBCT measurement.Conclusion:Clinical direct measurement and CBCT measurement were largely consistent in the evaluation of the alveolar bone height and width after the alveolar ridge preservation using deproteinized boving bone mineral (DBBM,Bio-Oss)and bioabsor-bable collagen membrane (Bio-Gide)in periodontal compromised molar sites of severe bone defects.

OBJECTIVE: To study the relationship between the radiotherapy resistance and autophagy. To provide a theoretiacal basis for drugs that regulate autophagy to improve radiotherapy sensitivity. METHOD: Flow cytometry (FCM) was performed to analyze the distribution of the cell cycle of CNE2 and CNE2/DDP cells under the action of X radiation. The expression of autopagy-specific gene Beclin1 and microtubule-associated protein light chain 3β (MAPLC3β) in CNE2 and CNE2/DDP cells was determined by real time PCR and Immumofluorescence staining. RESULT: CNE2/DDP and their parental CNE2 cells produced the G2-M phase arrest under the action of X radiation. With the radiation dose increasing,The cells which in the G2-M phase were more and more (P<0. 05). The G2-M phase arrest in CNE2/DDP cells was more obvious than in CNE2 cells (P<0. 05). The expression of Beclin1 and MAPLC3β in CNE2 and CNE2/DDP cells increased under the action of X radiation. What's more, the raise was more and more obvious with the increase of the irradiation dose(P<0. 05). The expression levels of Beclin1 and MAPLC3β in CNE2/DDP was lower than that in CNE2 cells (P<0. 05). CONCLUSION Autophagic cell death may be the one manner of death in nasopharyngeal carcinoma CNE2 and CNE2/DDP cells under the action of X radiation. The radiation resistance of CNE2/DDP cells may be related to the low expression of autophagy-related genes.
Apoptosis Regulatory Proteins ; genetics ; Autophagy ; Beclin-1 ; Carcinoma ; Cell Cycle ; Cell Line, Tumor ; radiation effects ; Dose-Response Relationship, Radiation ; Humans ; Membrane Proteins ; genetics ; Microtubule-Associated Proteins ; genetics ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; Radiation Tolerance ; X-Rays

Objective To analyze the current situation and problems of the international service trade of private medical institutions of Traditional Chinese Medicine (TCM) in Beijing,and provide the basis for the relevant policies of the international servicetrade of TCM.Methods Combining the stratified samplingandtheconveniencesampling,103 private medical institutions of TCM in Beijing were investigated by questionnaire,and the survey data were analyzed.Results 66.99％ of the visiting institutions carried out international trade in services of TCM,and 1 institution had set up branches abroad.2.90％ institutions of which have carried out international trade of TCM were preparing for overseas branches,and 8.70％ planned to establish overseas branches within 5 years.Conclusions The private medical institutions of TCM developerapidly,but change frequently.Private medical institutions lack compound talents and the enthusiasm of the trade in service.The construction of laws and regulations on trade in services of TCM by the government need improvement.The trade barrier of foreign countries on TCM is increasingly higher.

OBJECTIVE:To provide reference for promoting the rational use of essential drugs in primary medical institutions. METHODS:Twenty-three rural primary medical institutions(8 township health centers,15 village clinics)were randomly selected from 2 poverty-stricken counties in Chongzuo city of Guangxi Zhuang Autonomous Region. The utilization of essential drugs was investigated and analyzed statistically through exporting hospital information system data and on-site interviews. RESULTS:The utilization rate of essential drugs in sample township health centers was 72.4％,and the amount of essential drugs accounted for 78.3％. The utilization rate of variety from National List of Essential Drug was 36.9％,which in the Autonomous Regions Essential Drug List Supplement was 47.2％(utilization rate of chemical drugs was 53.9％,and that of Chinese patent drugs was 39.7％). The utilization rate of essential drugs in sample village clinics was 64.4％;utilization rate of variety from National List of Essential Drug was 18.3％, which in the Autonomous Regions Essential Drug List Supplement was 10.5％. The amount of 5 major categories accounted for 47.9％ of total amount of chemical agents,among which that of antimicrobial agents took up the highest proportion,being 35.9％. Among top 10 drugs in the list of amount,there were 8 national essential drugs and 4 antimicrobial agents. There were 2.90 kinds of drugs in each outpatient prescription averagely in sample township health centers;the average proportion of antibiotic prescription was 43.0％;the cost of each prescription was 33.12 yuan in average. CONCLUSIONS:The utilization rate of essential drugs in primary medical institutions is lower than the requirements of policy. There are great difference in utilization rate between chemical agents and Chinese patent drugs of township hospital in Autonomous Regions Essential Drug List Supplement. The cost proportion of antimicrobial agents is high. The rationality of prescription drug use still needs to be improved. It is suggested to guarantee essential drugs supply,adjust Autonomous Regions Essential Drug List Supplement in time,control the price of main categories and strengthen antimicrobial agent use monitoring so as to further promote rational drug use.