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 changes of programmed death receptor-1 (PD-1) in chronic hepatitis B (CHB) patients with different baseline of hepatitis B virus (HBV) DNA levels after treatment with adefovir dipivoxil (ADV).Methods One hundred CHB patients with positive hepatitis B e antigen (HBeAg),1 × 104 copy/mL≤HBV DNA≤1 × 107 copy/mL,and positive human leukocyte antigen-A2 were divided into two groups according to the baseline HBV DNA level:47 cases in low virus load group whose HBV DNA level was ≤1 × 105 copy/mL; 53 cases in high virus load group whose HBV DNA level was＞1 × 105 copy/mL.Both groups were treated with ADV 10 mg/d.Serum HBV DNA,HBeAg seroconversion rate,alanine aminotransferase (ALT) and total bilirubin (TBil) levels of both groups before treatment and 12 months after treatment were compared.Flow cytometry was used to test peripheral blood HBV-specific cytotoxic T lymphocyte (CTL) surface PD-1 and peripheral blood HBV-specific CTL level.Categorical data were tested by x2 test; quantitative data was compared with t-test.Results Peripheral blood HBV-specific CTL surface PD-1 of CHB patients in low virus load group was 20.17 ％±1.69％,which was lower than that in high virus load group (41.38％±2.30％,t =53.02,P＜0.01) ; peripheral blood HBV specific CTL levels in two groups were 0.37％±0.02％ and 0.17％± 0.02％,respectively (t=50.47,P＜0.01) ; ALT and TBil levels in low virus load group were both lower than those of high virus load group (t=13.07,P＜0.01; t=5.06,P＜0.01).Twelve months after treatment,HBV DNA of 25 cases (53.2％) in low virus load group and 10 cases (18.9％) in high virus load group were lower than the detectable level (HBV DNA＜500 copy/mL,x2 =12.89,P＜0.01);HBeAg seroconversion was achieved in 15 cases(31.9％) and 1 case (1.9％),respectively (x2 =16.72,P＜0.01) ; peripheral blood HBV-specific CTL surface PD-1 expression levels were 9.00 ％ ±1.38 ％ and 29.40 ％ ± 3.76 ％,respectively (t =36.80,P＜ 0.01) ; peripheral blood HBV-specific CTL levels were 0.65％±0.10％ and0.48％±0.07％,respectively (t=9.61,P＜0.01).Conclusions After treatment with ADV,along with the decrease of HBV DNA load,HBV-specific CTL surface PD-1 expression decreases,while HBV-specific CTL level increases.The changes in low virus load group are much more remarkable.

Objective To explore the correlation of E-cadherin(E-cad) mRNA expression with tumorigenesis and development of lung cancer.Methods The relative quantitative nested RT-PCR was used to detect the expression of E-cad mRNA in cancerous tissues,adjacent and distal tissues of tumor from 50 lung cancer patients,and non-cancerous benign lung tissues from 5 lung disorder patients.Results The means of expression rate of E-cad mRNA in cancerous tissue,adjacent and distal tissues of tumor,and non-cancerous tissues were 1.33?0.53,1.65?0.60,2.01?0.46 and 2.09?0.09 respectively.The data were evaluated by analysis of variance.There were significant difference among cancerous tissue,adjacent and distal tissue(F=18.810,P0.05).Also,there were no significant differences among squamous cell carcinoma,adenocarcinoma and adenosquamous carcinoma in the group of cancerous,adjacent and distal tissue.Conclusion The reduced expression of E-cadherin mRNA may play important role for tumorigenesis and development of lung cancer,and there was no evident relationship of E-cad mRNA with different pathological types of lung cancer.

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.

Objective To analyze and compare the advantages and disadvantages in high-field MRI scan and LAVA enhanced in the display of hepatic carcinoma capsule,in order to improve the early diagnosis and differential diagnosis level of primary hepatic carcinoma by MRI.Methods MRI data of 233 patients of primary hepatic carcinoma were retrospective analysed by two radiologists. Results 233 cases of primary hepatic carcinoma,except for 18 cases of diffuse hepatocellular carcinoma,a total of 239 lesions (54 small hepatocellular carcinoma,76 nodular hepatocellular carcinoma,109 massive hepatocellular carcinoma )were found .Hepato-cellular carcinoma capsule display rate was 139/239(58.16%).119 T1 WI,87 cases were found in T2 WI,and 139 cases were found in LAVA enhanced scan.25 lisions showed complete capsule on T1 WI,12 lisions showed complete capsule on T2 WI,59 lisions showed complete capsule on LAVA enhanced scan.Small hepatocellular carcinoma displayed capsule 21/54 (38.9%),nodular hepa-tocellular carcinoma 53/76 (69.7%),massive hepatocellular carcinoma 65/109 (59.6%).Conclusion High-field MRI conventional scan and LAVA enhenced scan can display PHC capsule better,LAVA enhanced (portal phase + delay phase)showed PHC capsule better than T1 WI and T2 WI.