0.98); immature cells would display in the WBC histogram when in higher proportion. Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to checked with microscopy for DC before reported.

Astrocytoma ; pathology ; Brain Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Ganglioneuroma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; pathology ; Oligodendroglioma ; pathology ; Synaptophysin ; metabolism ; Vimentin ; metabolism ; Young Adult

Objective To investigate the effect of great omentum combined with medical obturation glue on preventing thoracic cavity anastomotic leakage.Methods From August 2008 to September 2012,560 patients with esophageal gastric cardial carcinoma were enrolled and divided into two groups:the regular group (n =280) and the experimental group (n =280).In the regular group,anastomosis was reinforced with interrupted mattress sutures after esophageal gastric anastomosis was stapled.In the experimental group,anastomosis was covered with great omentum and medical obturation glue was sprayed to conglutinate after reinforced with interrupted mattress sutures.After that,gastric corpus was fixed upon the thoracic aorta and posterior chest wall.The clinical effects of the two groups were compared.Results Intrathoracic anastomotic leakage occurred in 8 cases (2.86％(8/280)) of the regular group,including 7 cases with symptomatic leakage and 1 case with asymptomatic loculate leakage.Seven patients were cured with conservative treatment and 1 patient with severe infection left hospital without cure.Average length of hospital stay was (55.6 ± 30.5) days postoperatively.Anastomotic stenosis occurred in 11 patients (3.93％,11/280).In the experimental group,one patient (0.36％,1/280) with asymptomatic loculate leakage was hospitalized for 20 days,and finally cured and discharged.8 cases with anastomotic stenosis occurred in the experimental group (2.86％,8/280).There was statistic difference in the rate of intrathoracic anastomotic leakage between the two groups (P =0.044),but there was no statistic difference in anastomotic stenosis between the two groups (P =0.484).Conclusion The technique of great omentum combined with medical obturation glue for preventing thoracic cavity anastomotic leakage,which is easy to perform,can obviously decrease the occurrence and attenuate the symptom of intrathoracic anastomotic leakage,and anastomotic stenosis increases unobviously.It also can shorten the length of hospital stay and is worthy of clinical promotion.

0.99). DC: reproducibility was good for neutrophils, lymphocytes monocytes, eosinophils and basophils. Comparison of the results by instrument with manual for normal samples in morphology, the correlation was better for neutrophils, lymphocytes and eosinophils (r:0.968～0.983) ,good for monocytes(r=0.917), not good for basophils(r=0.659);The WBC scattergram would change and alarm flags would display when there are neutrophilic stab granulocytes, abnormal or atypical lymphocytes and immature cells in higher proportion.Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to check with microscopy for DC before reported when it were doubtted.

Objective To evaluate the performance of the ABX Micro C-reactive protein(CRP)in determination of CRP.Methods The analytic characteristics including precision,carry-over,linearity, stability,interference and comparability were examined.Results The coefficient of variation(CV)was less than 5.1%,10% and d.3% for within-run,between-run and between-day,respectively.Carryover was less than 1.2%.Whole blood samples held at either room temperature or 4℃ were stable for 48 hours with relative deviation less than 6.0% relatively.Linear range was 1.0-70.0 mg/L using undiluted samples.The comparison between the ABX Micro CRP and Behring Nephelometer Ⅱ was well correlated Both serum:Y=0.996 7X-0.398 5,r~2=0.965 9;serum for BN Ⅱ,whole-blood samples for the ABX Micro CRP:Y=0.908 8X-0.138 2,r~2=0.959 4;both serum and whole-blood samples for the ABX Micro CRP: Y=1.001 7X-0.898 2,r~2=0.952 7.No obvious interference was observed by hyperhemoglobinemia and hyperlipidemia.Conclusion The determination of CRP test with ABX Micro is accurate and reliable.

Objective To construct an eukaryotic expression vector with human adipose most abundant gene transcript 1 (APM1) gene,and to investigate the transfection and expression of pCDEF-APM1 eukaryotic expression plasmid in HEK293 cells.Methods pCDEF-APM1 eukaryotic expression plasmid was constructed by DNA recombinant method.Expression vector pCDEF-APM1 was transfected into HEK293 cells with Effectene reagent.The level of human adiponectin protein in the supernatant of cell culture media was detected with double antibody sandwich ELISA.Results The sequence of DNA fragment from constructed pCDEF-APM1 plasmid was identical to that published in GenBank.There was raised human adiponectin protein level in culture supernatant of HEK293 cells tnmsfected with pCDEF-APM1.Conclusion The pCDEF-APM1,an eukaryotic expression plasmid for APM1 gene is successfully constructed.High protein expression of adiponectin can be obtained in HEK293 cells transfected with pCDEF-APM1 eukaryotic expression plasmid.

Dengzhan Shengmai capsule, as a compound Chinese patent medicine, consists of four herbs: Herba Erigerontis, Ginseng, Ophiopogon, and Schisandrae Chinensis Fructus, and contains significant components of flavonoids, lignans, saponins, and organic acids. It is widely used clinically to treat cerebrovascular diseases such as chronic cerebral hypoperfusion and dementia with remarkable efficacy. This study proposes a research strategy for multi-component traditional Chinese medicine metabolites based on prediction databases and unfolds the analysis using Dengzhan Shengmai capsule as an example. Using the UPLC-Q-TOF/MS method, the analytical method was established and detected biological samples such as urine, feces, and bile of rats before and after administration based on the prediction of theoretical metabolites of Dengzhan Shengmai capsule. The possible secondary fragment ion information of metabolites was identified by comparing the detected results with prediction databases. The metabolites were identified based on the archetypal component mass spectrometric cleavage law and multistage mass spectrometric data. 51 metabolites, mainly flavonoid, organic acid, and lignan constituents, were finally identified from rat biosamples based on 306 theoretical metabolites of Dengzhan Shengmai capsule. This study provides a new strategy for the identification of metabolites in vivo and the analysis of metabolic pathways of TCM. The study complied with the procedures established by the Animal Experiment Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and passed the animal experiment ethics examine (No. 00003645).

This study was purposed to investigate the efficacy of different whole flow lysis reagents for lysis of red blood cells in flow cytometric analysis. The expression of immunocytes was detected by flow cytometry after lysis of red blood cells using commercial reagents (Optilyse C, FACS Lysing Solution) and self-made red blood cell lysis reagents (RBC Lysis Buffer), the detection results were analyzed comparatively. The results showed that there was no significant difference in the percentage of CD3e(+), CD3e(+)CD4(+), CD3e(+)CD8a(+), CD3e(-)CD19(+), CD3e(-)NK1.1(+) and Gr-1(+) cells between 3 different lysis reagent groups. However OptiLyse C solution was suitable to Gr-1(+) cell detection, but did not suit to Foxp3(+) Treg detection. The self-made RBC Lysis Buffer and FACS Lysing Solution were suited to Foxp3(+) Treg detection. It is concluded that the use of self-made RBC Lysis Buffer for flow cytometry can get the lysis efficiency of commercially available lysis solutions when samples are prepared in accordance with standardized procedure. The self-made RBC Lysis Buffer not only can satisfy experimental requirements, but also can reduce the experimental costs.
Animals ; Erythrocyte Count ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; instrumentation ; methods ; Immune System ; immunology ; Indicators and Reagents ; analysis ; Mice ; Mice, Inbred C57BL

Objective To investigate the effects of different Leptospira strains on phagocytosis of peritoneal macrophages of guinea pigs,and explore the role of innate immune in the pathogenesis of leptospirosis. Methods Peritoneal macrophages of guinea pigs were infected in vitro by three different Leptospira strains,the virulent Leptospira interrogans serovar Lai type strain Lai,the avirulent L.interrogans serovar Lai type strain IPAV,and the nonpathogenic L.biflexa serovar Patoc type strain PatocⅠ,respectively,and heat inactivated Staphylococcus epidermidis was added 0.5,1.5,3 and 6 h after infection and incubated for 30 min.The effect of Leptospira on the phagocytosis of macrophage was evaluated by the inactivated Staphylococcus epidermidis phagocytosis rate and phagocytosis index.Phagocytosis and ultrastructure of peritoneal macrophages were observed by transmission electron microscopy 3 h after infection,and changes of cytoskeleton of the macrophages were observed by laser scanning confocal microscopy. Results The phagocytic rates and phagocytic indexes of strain Lai,strain IPAV and strain PatocⅠinfection groups were significantly lower than those of control group 3 h and 6 h after infection(P

OBJECTIVETo express the L1 protein of human papillomavirus type 16 (HPV16) in insect cell suspension culture system.

METHODSOptimized the conditions of suspension culture, recombinant virus amplification and protein expression. Determined the virus tilter by plague analysis and detected the target protein by SDS-PAGE and Western blot; The formation of VLPs by HPV16 L1 protein was observed with TEM.

RESULTSThe Sf9 cells could grow better in suspension culture with seeding density of 5 x 10-5 cell/mL and the maximum expression quantity was obtained by infection of cells with rBacV/HPV16L1 (MOI =10) and harvesting after 72-84 h. HPV16L1 protein could assemble into VLPs in Sf9 cells observed with TEM.

CONCLUSIONThe conditions of cell culture, virus amplification and protein expression were optimized. HPV16 L1 protein could assemble into VLPs in Sf9 cells, which would provide a foundation for further study of the vaccine and diagnosis kits.

Animals ; Capsid Proteins ; biosynthesis ; Cell Proliferation ; Oncogene Proteins, Viral ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Spodoptera ; Suspensions ; Virion ; ultrastructure