Objective To construct pET21a-sBAFF by cloning the extracellular regions of 134-285 amino acids of BAFF, a member of human TNF family, and then express the gene in prokaryotic cells and purify the expressed product.Methods cDNA of K562 cell line was used as the template to amplify sBAFF gene to construct pET21a-sBAFF.Expression of sBAFF in E.coli BL21 was induced by IPTG, and the expressed proteins were assayed by SDS-PAGE.The bacteria were analyzed by sonication, and the target proteins mainly existed as inclusion bodies.Then sBAFF was purified by Ni2 +-IDA affinity chromatography.SDS-PAGE electrophoreses displayed that the expressed sBAFF migrated with a relative molecular weight of 18000.ResuIts The induction parameters such as temperature and inducing time were optimized.The target protein accounted for 38.59%of the total bacterial proteins.After refolding, 38.14% of sBAFF proteins were polymerized as an active trimer.The dimer form of sBAFF, which is representative of wrongly refolded product, accounted for very few.ConcIusion The expression and purification of BAFF which formed active trimer after refolding pave the way for its further function study and provide a novel approach for the development of BAFF-targeted therapeutics for autoimmune diseases.

Objective:To evaluate and compare the identification of several DNA barcoding candidate sequences on Illicium difengpi and its fake I. jiadifengpi. Method:Samples from different origins of I. difengpi and I. jiadifengpi, were collect extraction of total DNA,nuclear gene ITS2 sequence,chloroplast rbcL,matK gene sequence were selected for PCR amplification,product purification and sequencing,and CondonCode Aligner V3.7.1 was used to proofread stitching. Result:PCR amplification and sequencing of rbcL sequences of I. difengpi and I. jiadifengpi were not satisfactory. It is assumed that their rbcL sequences were too long with slow evolution,which was unsuitable for interbreeding. The success rate of matK sequencing of I. difengpi and I. jiadifengpi was 0 and 76.8%,which may be because primer standards were different for matK sequences of different groups. The results of PCR amplification and sequencing of ITS2 on I. difengpi and I. jiadifengpi were successful,with the success rate of sequencing was 89.3% and 91.2%. Analysis sequencing results, the total length of ITS2 sequences was 268 bases,and there were 2 variation sites of I. difengpi. The total length of ITS2 sequences was 430 bases,and there were 4 or 3 variation sites of I. jiadifengpi. It shows that ITS2 sequences of I. difengpi and I. jiadifengpi were short and has obvious variability and can be amplify,that ITS2 sequence was better than rbcL and matK sequence in molecular identification of I. difengpi and I. jiadifengpi. Conclusion:DNA barcoding based on ITS2 sequence was a powerful and efficient tool for identification of I. difengpi and its fake I. jiadifengpi.

Objective To analyze relationship between dual-source MDCT and pathology in renal cell carcinoma.Methods 129 patients with pathology were proved renal cell in our hospital from 2009 to 2012.According to the latest 2004 WHO pathological classification,CT features of renal carcinoma were compared with surgical and pathological results.Results The enhancement de-gree of lesions on contrast CT was correlated to the modal of renal malignant cells’ranking.There was no certain correlation be-tween integrity on the edge of the tumor in CT and pathological tumor capsular.The short-hair sign surrounding the margin of tumor strongly indicated renal capsule invasion (P <0.01).Agreement between CT-Robson staging and surgical-pathologic staging was good(Kappa=0.75).Conclusion The CT finding of renal cell carcinoma is correlated with tumor cell characteristic and inter-nal structure.Dual-source MDCT has important clinical value in the diagnosis of renal cell carcinoma.

Objective To investigate the therapeutic effect of intravascular embolization of carotid-cavernous fistulation.Methods 5 cases were examined by DSA of brain to find out the sites of fistulation.The detachable balloon or GDC(Guglielmi Detachable Coil)were employed in embolotherapy.Results 1 cases were embolized successful with detachable balloon and kept free circulation of ICASs;The embolism of ICA involved side were performed in 2 cases for patient with pseudoaneurysm and 1 patients cavernous involved were embolized with GDC for small mouth of the fistula.Another case balloon raptured two times when it was detached to cavernous after one free balloon was dept in cavernous,the patient didn't want to embolized the ICA involved,the trentment was stoped,the patients symptoms was disappeated after one week of therapy.Conclusion The site and size of carotic-cavernous fistulation can be detected by DSA of brain intravascular embolization is the first-choise method of therapy for it.

BACKGROUND: The ideal biomaterial means absence of cytotoxic effect and immunological rejection, degradation at right moment, and a well histocompatibility. Whether bio-derived bone can be used in vivo for long time and exerts functions deserves to be studied.OBJECTIVE: To investigate the local histocompatibility after bio-derived bone implanted into mouse and the effect on immunofunctions.DESIGN: A randomized controlled animal experiment. SETTING: Key Laboratory of Pathobiology, Ministry of Education, School of Basic Medical Sciences, Jilin University. MATERIALS: This study was performed at the Key Laboratory of Pathobiology, Ministry of Science, School of Basic Medical Sciences, Jilin University from March to July in 2006. Eighteen BALB/C mice (weighing 20±2 g, half male and half female), one male Kunming mouse (weighing 20 g), and one female rabbit (weighing 2.5 kg) were included in the experiment. All the experimental animals were provided by Laboratory Animal Center, School of Basic Medical Science, Jilin University. The disposal of the experimental animals in the test process accorded with ethical guidelines for the use and care of animals. Porcine cancellous bone (iliac bone) was purchased from the market. Iscove's Modified Dulbecco's Medium (IMDM, Hycolone, USA), fetal bovine serum (FBS, Gibco Co., Ltd, USA), methyl thiazolyl tetrazolium (MTT, Sigma, USA), and concanavalin A (ConA, Sigma Co., Ltd, USA) were used.METHODS: Bio-derived bone was prepared from commercial porcine bone. ① Eighteen BALB/C mice were randomly divided into three groups with 6 mice in each group: a control group (simple local muscle injury without implantation), a bio-derived bone implantation group ( implanting bio-derived bone into the lower limb), and a xenogenic bone implantation group (femoral bone from Kunming mouse was implanted into the muscle of lower limb). ② Twenty-one days after operation, the implant material and surrounding tissue were obtained for gross observation and haematoxylin-eosin staining to investigate the histocompatibility of bio-derived bone. Mouse immunofunction was assessed by complement-mediated cytotoxicity test. Absorbance was determined with an automatic ELISA reader at 570 nm to assess the cytotoxicity.MAIN OUTCOME MEASURES: ①Histocompatibility of implant surrounded tissue. ②Lymphocyte stimulation indices after induction of concanavalin A. ③ Cytotoxicity in each group after complement-dependent cytotoxicity test.RESULTS: Eighteen BALB/C mice were included in the final analysis. ①Histocompatibility of implant surrounded tissue: In the bio-derived bone implantation group, 21 days after bio-derived bone implantation, there were no presentation of congestion, degeneration, necrosis and diapyesis around the implant in gross, plenty of fibrous connective tissue invaded into the pores of the bio-derived bone, encapsulation and forming the fibrous capsule. A great quantity of neutrophils and macrophages were not detected around the implant by haematoxylin-eosin staining. Bio-derived bone was encapsulated with fibrous tissue, and part of the biomaterial began to degrade, and being replaced with fibrous tissue. Regarding xenogenic bone implantation group, necrotic tissue was detected in the cross-section of the muscle in gross. A lot of neutrophils, macrophages and necrotic tissue were detected around the implant by haematoxylin-eosin staining. ②Lymphocyte stimulation indices: The stimulation index of xenogenic bone implantation group was significantly larger than that of control group (P ＜ 0.05). There was no statistical difference between the bio-derived bone group and the control group (P ＞ 0.05). ③Cytotoxicity: The cytotoxicity of xenogenic bone implantaion group was significantly larger than that of control group (P ＜ 0.05). There was no significant difference in cytotoxicity between the bio-derived bone implantation group and the control group (P ＞ 0.05).CONCLUSION: The obtained bio-derived bone causes little immunoreactions, has no obvious cytotoxicity or inflammatory reactions, and possesses a good histocompatibility and bio-safety.

OBJECTIVETo research on the clinical application of the WHO 1979, 1993 and 2000 classifications of tumours of the central nervous system.

METHODSWith the analysis of 4 373 cases, problems were studied in applying the WHO classifications.

RESULTS(1) The 2000 classification used double criteria of WHO and ICD-O. The relationship between them was very confusing. (2) Opinions suggested for about 15 types of the tumours of meningothelial cells. (3) It was difficult to separate benign, atypical or anaplastic meningiomas. (4) The diagnostic criterion of anaplastic gliomas was not clear. (5) Practically, glioblastoma was a type of mixed gliomas. (6) More cases should be studied in order to recognize special types of the tumours of neuroepithelial tissue.

CONCLUSIONSSuggested to make out a criterion of classification and grading that is more applicable for clinical diagnosis.

Objective To explore the value of dual-source CT spectrum energy curve and iodine quantification in determining the degree of differentiation of clear cell renal carcinoma(ccRCC).Methods The radiographic data of 38 cases with pathologically confirmed ccRCC were analyzed retrospectively.All patients underwent dual-source CT,cortical phase and nephrographic phase dual energy scan,including 22 well-moderated differentiated(gradeⅠ,gradeⅡ)and 16 poorly differentiated neoplasms(gradeⅢ,gradeⅣ).The slope of spectrum energy curve (40-70 keV,70-100 keV,100-140 keV)and the normalized iodine concentration (NIC)between the two grades were compared. Results In the spectrum energy curve 40-70 keV and 70-100 keV segment interval,the slope of spectrum of well-moderated differentiated group was higher than that of poorly differentiated group,the difference was statistically significant(P<0.05).In the 100-140 keV segment interval,the slope of spectrum of well-moderated differentiated group was higher than that of poorly differentiated group, but the difference was insignificant(P>0.05).The NIC of well-moderated differentiated group was significantly higher than that of poorly differentiated group(P<0.05).Conclusion The slope of spectrum energy curve and NIC are correlated with the differentiation degree of ccRCC.The slope of spectrum energy curve and iodine quantification of dual-source CT could be used as new methods for preoperative grading of ccRCC.

Objective To investigate the clinical preemptive analgesic efficacy of parecoxib sodium(PS) at different administration timing in the patients with total hip arthroplasty(THA).Methods Sixty patients receiving THA were prospectively enrolled and randomized into three groups.The group A started to be intravenously injected by PS 40mg/d on preoperative 3 d until operation day;the group B was intravenously injected by parecoxib sodium 40mg at preoperative 30 min;the group C began to be intravenously injected by the same dosage of normal saline at the same time point as the group A.The rest pain was assessed by using the visual analog scale(VAS) at postoperative 6,24,48,72 h.The duration of patient-controlled intravenous analgesia(PCIA) and total dosage were recorded.The first time unaided ambulation time was observed.Results The VAS scores at various postoperative time points in the group A and B were significantly lower than those in the group C(P＜0.05),the VAS scores at postoperative 6,24 h in the group A were remarkably lower than those in the group B.The PCIA duration in the group A,B and C were 25.05±10.32),(36.75± 13.91),(50.40 ± 15.17)h,respectively,the pair-wise comparison of the group A,B and C showed statistical difference(P＜0.05).The total dosages of PCIA drug in the group A,B and C were(29.25 ± 4.58),(34.50 ± 5.09),(62.65 ±10.52)tg,respectively,the dosage in the group A and B was significantly lower than that in the group C(P＜0.05).The first time unaided ambulation time in the group A,B and C were (2.75 ± 0.81),(3.05 ± 1.08),(4.10 ±-0.92)d respectively,which in the group A and B was earlier than that in the group C (P＜0.05).Conclusion Continuously using PS on preoperative 3 d can increase the analgesic effect in THA patients and is conducive to the functional rehabilitation and increase the patient satisfaction.

Objective To learn 2009~2014 Shaanxi Province sentinel surveillance six classes of HIV infection focus groups, and estimates of HIV-1 new infection.Methods Used enzyme-linked immunosorbent assay (ELISA)and Western blot (WB)experiments for the 2009~2014 HIV sentinel surveillance Shaanxi Province Category 6 focus groups conducted a total of 77 778 HIV antibody screening and confirmatory testing estimates of HIV-1 new infection.Results 2009~2014 men who had sex with men and people with HIV infection rate were 3.75%,8.77%,3.50%,5.00%,6.20% and 5.75%,and a slow upward trend;HIV-1 new infection were 5.04%,8.96%,5.01%,5.95%,4.68% and 6.39%,the overall downward trend. Young students,drug addicts,sex workers,pregnant women,and male STD clinic attenders five people with HIV infection and HIV-1 new infection were emerging to remain low.But male STD clinic attenders of HIV infection and HIV-1 new in-fection was emerging slowly rising trend.Conclusion Shaanxi MSM HIV infection and HIV-1 new infection were high,but HIV-1 new infection had decreased slowly.Emerging trend should continue to increase the population of the intervention ef-forts.The infection rate in other monitoring population was relatively low but a few people on the rise,the need to take the necessary coping methods.

Objective To establish and optimize three-cube FRET assay in living cells and analyze subunit assembling of iGluR receptors. Methods Taking HEK293 cells cotransfed with pECFP and pEYFP as negative control, and those transfected with pECFP-YFP as positive control,different calculation methods using fluorescence microscopy were compared. Results These calculation methods were all suitable for FRET measurement in the system. but the measurement results were affected by the ratio of Donor/Acceptor (D/A) in some degree,and different calculation methods have different optimized conditions. FRET measurement using FR value showed subunit specific assembly of iGluR subtypes.Conclusion There are different optimized conditions for these different calculation methods in the three-cube FRET measurement system,and a further evidence is provided for subunit specific assembling of iGluR subtypes from the FRET assay.