Objective To investigate the neuroprotective effect of ginkgolide B (BN52021) o n the histopathology of brain tissue after traumatic brain injury in rats. Methods Forty-eight healthy SD rats, weighing 250 g, were evenly randomized into 4 groups: sham control group, model group, low dose BN52021 group and high dose BN52021 group. Rats in the latter 3 groups were made into fluid percussion brain injury models. After operation, the rats in the low and high dose BN52021 groups were treated with BN52021 (low dose: 1 mg/kg, ip, high dose: 5 mg/kg, ip, once daily for 7 days). On the 7th day after treatment, cerebral tissues were harvested from each group, and the histopathological changes of brain tissue were observed by Fast blue, electron microscope and immunohistochemical method. Results Compared with sham control group, model group had significantly decreased neurons (P<0. 05), increased OX-42 immunoreactive microglial cells and astrocytes (P<0. 05), and cells positive for caspase-3 (P<0. 05). Electron microscope found chromatin aggregation, nuclear fragmentation, rounder and larger mitochondria, void formation and disappeared cristae of mitochondria, endoplasmic reticulum hypertelorism, increased lysosomes, and nuclear membrane folding. Compared with model group, the low and high dose BN52021 groups had significantly decreased proportions of microglial cells and astrocytes (P<0. 05), significantly decreased caspase-3 positive cells (P<0. 05), and improved ultrastructure, with the improvement in the high dose group being more notable than that in the low dose group. Conclusion BN52021 has protective effect on the morphology of brain tissue in rats with traumatic brain injury.

Objective To evaluate the performance of KRAS gene mutation detection in 2014 external quality assessment ( EQA ) program and discuss the problems in clinical laboratories .Methods The sample panel of 2014 EQA program contained 5 artificial formalin-fixed, paraffin-embedded ( FFPE) samples.The participating laboratories were asked to report their results before the deadline .The scores of EQA and the rate of overall coincidence , false positive and false negative were calculated .Results The EQA program for KRAS testing was set twice a year .In 2014, 58 and 57 valid lab results were submitted respectively.About 79.31%(46/58)and 94.73%(54/57) of the laboratories were correct for all samples. The coincidence rate of positive samples were 93.53% ( 217/232 ) and 96.49% ( 165/171 ) . The coincidence rate for negative ones were 100%(58/58) and 98.25% (112/114).The false-negative ratio was 1.29%( 3/232 ) and 0%.The false-positive ratio was 4.14% ( 12/290 ) and 3.15% ( 9/285 ) . Conclusions The results of 2014 EQA for KRAS gene mutation testing suggested that the performance of laboratories had been improved significantly , however , the false-negative and false-positive results had always been the major problems affecting the accuracy of KRAS mutations testing .Laboratories needed to standardize the testing process and manufacturers should optimize the reagents and the way of interpretation , to guarantee the performance of KRAS gene mutation detection .

Objective To investigate the infection of Chlamydia (CT) Untie urea mycoplasma (UU) and person type mycoplasma (MH) in urology outpatients clinic and sexually transmitted diseases clinic and drug resist-ance. Methods A British UNIPATH Clean View Chlamydia rapid immunoassay kits and Tianyang Zhongshan City electronic biosensor Limited production of mycoplasma culture and identification of susceptibility kit were used for the detection of secretion from 3,280 cases of non-gonococcal urethritis (GNU). Results Of 3,280 cases of GNU, only 241 cases were detected with positive CT, accounting for 7.35%, 1163 cases with simply UU positive, ac-counting for 35.46%, only 14 cases with MH positive, accounting for 0.43% ,. Overall detection rate of UU and CT was 42.77% and 11.59% respectively. 122 cases were detected with positive UU + MH, accounting for 3.72%. Female infection rate was higher than that of males (P < 0.01 ) ; UU, MH, MH + UU were more sensitive to azithremycin, Josamycin, doxycycline, roxithromycin, Minocycline. Conclusion The infection rate of urogenital mycoplasma and ehlamydia is higher and drug resistance rate is different. Azithromycin and Josamycin are the best to treat mycoplasma infection, followed by doxycycline, Pyronine Doxycycline, Minocycline.

Circulating tumor DNA (ctDNA) has shown great potential in targeted therapy efficacy prediction, monitoring, high-risk population screening, differential diagnosis, minimal residual disease (MRD) monitoring, and prognosis prediction. The detection of specific gene mutations in ctDNA had been included in clinical practice guidelines for certain tumors to predict the drug efficacy and monitor resistance. A small number of approved companion diagnostic reagents have been used in clinical setting. However, the clinical validity of most ctDNA-related biomarkers it still in the research stage. Besides, the establishment and validation of laboratory-developed tests (LDT) are also problems that need to be solved urgently. Therefore, for the moment, the clinical application of ctDNA analysis is like two sides of a coin, with both opportunities and challenges.

Objective To construct the gall-stone patients' nursing workload measurement project by Delphi method,so as to measure single disease workload scientifically and reasonably.Methods The gall-stone patients' nursing workload measurement project were designed on the basis of consulting literature and experts discussion,then two rounds of consultations were made with experts by using the Delphi method.Results The valid recovery rate of questionnaires of two rounds consultations were all 100％ (22/22).The expert authority coefficient was 0.886 1.Nursing workload measurement project,directly related to the gall bladder calculi 54 nursing projects,including technical 46,unskilled 7;indirectly related to the gall bladder calculi 36 nursing projects,including technical 17,unskilled 19.Conclusions The measurement project of gall-stone patient determined by Delphi method has high reliability,which is significant to the scientific measuring of nursing single disease workload.

Objective To explore the expression change of Toll Like Receptor 4 (TLR4) in lung ischemia-reperfusion injury in rabbits and the influence of clemastine fumarate on it. Methods Fifty rabbits were divided into five groups randomly (n=10): Sham (N+S), reperfusion for 2 hours (N+I1/R2), reperfusion for 4 hours (N+I1/R4), clemastine fumarate + reperfusion for 2 hours (F+I1/R2) and clemastine fumarate+reperfusion for 4 hours (F+I1/R4). The ischemia time in each group was 1 hour and normal saline was given respectively in groups of N+S , N+I1/R2 and N+I1/R4. Western blotting , RT-PCR and immunofluorescence were used to detect the expression of TLR4 in lung tissue , and the changes of ultrastructure in ischemia-reperfusion lung tissue were observed by electron microscope. Result The expression of TLR4 was elevated obviously in ischemia-reperfusion lung tissue (P<0.05), and there was positive correlation between the increased TLR4 level and reperfusion time (P<0.05), the swelled and thick-ridge mitochondria were observed in type II alveolar epithelial cells after LIRI (P<0.05); but clemastine fumarate inhibited the expression of TLR4 in lung tissue significantly caused by LIRI (P<0.05). And the mitochondria injury was reduced in the groups of clemastine fumarate. Conclusion TLR4 expression is elevated in lung tissue after LIRI; clemastine fumarate inhibits the expression of TLR4 caused by LIRI and protects the lung tissue from LIRI in rabbits.

E2 gene of BVDV Changchun 184 strain was cloned and inserted into the shuttle expression plasmid vector pMV261,the recombinant shuttle plasmid pMV261-E2 was constructed.Then pMV261-E2 was transformed into BCG successfully and obtained recombinant BCG which was resistive to kanamyein.The recombinant BCG were identifieated by PCR.E2 gene expression in recombinant BCG was induced in 45℃,then the SDS-PAGE and western blotting was used to analyze the expression product.The results indicated the BVDV E2 gene was expressd in BCG successfully.

Objective To assess the relationship between the Chlamydia pneumonia(CPn)infection and coronary heart disease(CHD).Methods Blood samples from 200 CHD patients and 100 heathy controls were col-lected,and Cpn IgM and Cpn IgG were tested by ELISA.Results The Cpn IgM were found in 113 cases (56.5%) and in 24 controls(24.0%).The Cpn IgG positive cases were in 145(72.5%) patients and in 43 controls (43.0%).The positive rate of Cpn IgM and Cpn IgG in CHD group Was higher than that in control group(P<0.05).Conclusion:CPn positive rate is higher in CHD group than that in control group.Cpn is closely related to the pathogenesis of CHD.

BACKGROUND: There are different methods to isolate and culture human nucleus pulposus cells, and the differences in digestive enzymes components and digestion time quite are significant. So how to rapidly and efficiently harvest human nucleus pulposus cells has become a research hotspot. OBJECTIVE: To optimize the digestive enzymes components and digestion methods for the preparation of human nucleus pulposus cells. METHODS: Nucleus pulposus tissue specimens were selected from three adult discs in the Department of Orthopedics, China-Japan Union Hospital of Jilin University. The acute traumatic disc tissues that outstanding to the spinal canal were taken under aseptic conditions, and then the peripheral white annulus and jel y-like nucleus pulposus in the center could be seen. According to different mixed enzyme concentration ratio, the samples were divided into two groups. The enzyme Ⅰ group was treated with 0.2% Ⅱ col agenase; and the mixed enzymeⅡ group was digested with 0.25% trypsin for 30 minutes, and then treated with 0.2% Ⅱ col agenase. According to digestion time, each group was divided into three subgroups: 2 hours group, 4 hours group, and overnight group. Final y, suspended cel volume was decided as 2 mL to count cells. Dulbecco’s modified Eagle’s medium containing fetal bovine serum was used for cel culture in vitro. Trypan blue staining was performed to count total cel number and ratio of living cells. Methylthiazolyldiphenyl-tetrazolium bromide assay was used to detect the growth curve of nucleus pulposus cells. RESULTS AND CONCLUSION: Based on the two digestion enzyme concentration, the number of digested cells in the enzyme Ⅰ group was larger than that in the enzyme Ⅱ group after digested for 2 and 4 hours, but the difference was not significant (P > 0.05). Overnight, cellsurvival rate was decreased in the enzyme Ⅰ group after digested for 2 and 4 hours when compared with the enzyme Ⅱ group, and the difference was significant (P < 0.05). After digested for 4 hours, tissue blocks disappeared, and the number of cells reached maximum. The results indicate that enzyme Ⅰgroup composite with Ⅱ col agenase is benefit for the separation of nucleus pulposus cells, and the digestion time is appropriate to 4 hours. This condition has the advantages of simple operation, high efficiency and low cost, and it considered that digestion of nucleus pulposus tissues with 0.2% Ⅱ col agenase for 4 hours is the best condition to obtain nucleus pulposus cells.

BACKGROUND:Advancement in bioengineering based upon tissue engineering techniques may offer the possibility of repairing degenerative intervertebral disc.
OBJECTIVE:To summarize the research progress in the scaffolds of tissue engineered intervertebral disc.
METHODS:A computer-based retrieval was performed to search manuscripts describing tissue engineered intervertebral disc scaffolds published between January 1st, 1900 and December 31st, 2012 in PubMed database with the key words of“tissue engineering, intervertebral disc, scaffold”in English.
RESULTS AND CONCLUSION:Scaffold is an important part of tissue-engineered research. There are three kinds of materials for intervertebral disc scaffolds:natural biomaterials, synthetic materials, and composite materials. A variety of scaffold materials have their own advantages and disadvantages. Up to now, none of these scaffold materials is accepted as the most suitable one. The selection of scaffold materials is stil to be further studied. The study and development of nanoscale biomaterials is an inevitable trend. Otherwise, with the help of bionics, improving scaffolds is also an inexorable trend in progress of simulating human intervertebral disc. Furthermore, injectable scaffold is also an research hot spot, and the selection range of injectable scaffold materials mainly focuses on chitosan, typeⅡcolagen,hyaluronic acid,fibrin,elastin,and alginate.C urrently, studies on chitosan as a scaffold material are relatively more.