Objective To set up a mouse model of spondyloarthritis,analyzethe clinical phenotype,radiographic and pathological features,and investigate the therapeutic effect of cysteine-rich 61 (CCN1) monoclonal antibody in spondyloarthritis mouse model.Methods Proteoglycan from bovine nasal septum was used for immunization of 14-16 week old female BALB/c mice.CCN1 monoclonal antibody 093G9 or control immunoglobulin (Ig)G were injected to the spondyloarthritis mice.The arthritis scores were analyzed by t test.Peripheral and axial joints disease development was assessed by Micro-CT and histology.Results Proteoglycan immunized mice began to develop peripheral arthritis in the 8th week.The peripheral arthritis score reached the peak (10.5±1.5) in the 11th week,with the inflammation and spur formation of the ankle and knee joint.We found infiltration of inflammation cells in intervertebral discs of the lumbar vertebrae and the caudal vertebrae.Chondrocyte proliferation couldbe seen in the meniscus of knee and lumbar intervertebral discs.In the 18th week,the intervertebral discsof thoracic vertebrae and the cervical vertebrae were also damaged.Abundant chondrocytesgathered in the intervertebra] discs.The inflammation and new bone for-marion of peripheral and axial joints were more severe in control IgG group than 093G9 group.The peripheral arthritis score in the 093G9 group decreased significantly after 2 treatments,[(2.8±1.3) vs (4.2±2.1),t=2.516,P＜0.05].The difference in arthritis scores between the two groups was the most significant after 8.treatments,[(2.0±2.0)vs (5.3±2.0),t=4.082,P＜0.01].Conclusion The mouse model of spondyloarthritissimulates human spondyloarthritis,including inflammation and new bone formation in p()gheral and axial joints.CCN1 monoclonal antibody can improve the inflammation and new bone formation inspondyloarthritis mouse model.

Objective To compare the ROSCO disk diffusion method with broth microdilution method (CLSI, M27-A) for antimicrobial susceptibility test of Candida species isolated from patients with lung cancer. Methods Danish ROSCO company disk diffusion testing method and bio Merieux ATB FUNGUR2 were applied to test 5-flucytosine, fluconazole, itraconazole and amphotericin B antimicrobial susceptibility for 78 Candida species strains isolated from patients with lung cancer. Results Through evaluating the susceptibility to 5-flucytosine, amphotericin B, fluconazole and itraconazole by disk diffusion method, the Kappa value was 0.89. The sensitive strains detected by one method did not show resistance in another method. The sensitive rates of 78 strains of Candida species to 5-flucytosine, amphotericin B, fluconazole and itraconazole were 88.20 %, 89.17 %, 56.34 % and 52.12 %. The susceptibility of C.albicans, C.tropicalis, C.glabrata and C.krusei to four kinds of antifungal agents was 90.95 %, 85.71 %, 67.50 % and 41.67 %respectively. Conclusions Results of disk diffusion method coincide well with broth microdilution method. It can be chosen as a clinical routine method for antimicrobial susceptibility test.

Interleukin-21 is a type I cytokine mainly produced by activated CD4+ T cells that acts as a regulator of immune system. In this work, hIL-21cDNA was amplified from human peripheral blood lymphocytes by RT-PCR, and then inserted into pPIC9K. The recombinant vector pPIC9K-hIL21cDNA was linearized by Sac I, and transformed into Pichia pastoris strain GS115 by electroporation. Transformants were selected by G418 and confirmed by PCR. The recombinant protein was expressed and secreted into the supernatant after inducing by methanol. SDS-PAGE analysis indicated the molecular weight of rhIL-21 was about 16 kD. ELISA results show that the yield of rhIL-21 reach 229.28 mg/L, rhIL-21 was purified from culture supernatants, and it was purified to about 95% purity with ion-exchange chromatography. When co-stimulate with Con A, rhIL-21 can promote the proliferation of human lymphocytes. This is the first expression of bio-active rhIL-21 in Pichia pastoris. It lays a foundation for further research in immunotherapy and cancer therapy.
Electroporation ; Genetic Vectors ; genetics ; Humans ; Interleukins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; analysis ; biosynthesis ; genetics

Objective To explore the effect of therapeutic touch(massage)on the neuTobehavior of preterm infants.Methods Sixty preterm infants( gestafional age in 31-36 weeks),through natural delivery or caesarean section were randomly assigned to therapeutic touch group(n=30)and control group(n=30).Both groups re-ceived routine pharmacothempy and nursing care and followed up for half a year from discharge.Touch group re-ceived additional whole-body massage.All subjects were evaluated with neonatal behavioral neurological assessment (NBNA)in one month after birth-and mental development index(MDI), psyehomotor development index(PDI)in 3 months and 6 months after birth.Results The NBNA scores(P<0.01),as well as MDI and PDI(P<0.05)were significantly higher in therapeutic touch group than the control.Conclusion Therapeutic touch intervention im-proves nervous system of preterm infants.

Objective T9 provide candidate molecules for developing recombinant anti-idiotypic antibody (anti-Id) vaccine of gastric carcinoma by selection of recombinant anti-Id to monoclonal antibody ( McAb) MGb1 directed against the cancer with phage display technique.Methods Balb/c mice were immunized with MGb1 and the mRNA was isolated from the spleens of the immunized mice. The VL and VH cDNAs of the antibody were amplified separately by RT-PCR and assembled into ScFv DNAs with a linker DNA. The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage antibody ScFv library. After four rounds of biopanning to the library with MGb1, the MGb1-positive clones were selected from the enriched phages by ELISA. The types of the anti-Id ScFv displayed on the selected phage clones were preliminarily identified by competition ELISA. Results The VL and VH cDNAs was about 320 bp and 340 bp, respectively. The ScFv DNA were about 750 bp. After four rounds panning to the phage antibody ScFv library with MGb1, 18 MGb1-positive phage clones displayed anti-Id ScFv were selected from 50 pre-selected phage clones, among which 4 clones displayed β or γ type anti-Id ScFv. Conclusion The phagedisplayed anti-Id ScFvs to McAb MGb1 are successfully selected by recombinant phage antibody technique, which might lay a foundation for screening the anti-Id ScFv possessing the characteristics of inducing anti-gastric carcinoma immunity.

Through introducing and summarizing the application of multimedia technology in the process of biochemistry teaching,the article discusses the contribution of multimedia technology in teaching method,teaching organization,teaching content and so on and points out the importance of multimedia technology in biochemistry teaching.

Objective To clone the cDNA encoding human HMGB1, express it in E. coli, and identify its biological activity. Methods Human HMGB1 cDNA was amplified by RT-PCR and cloned into vector pUC19. After sequence analysis, the cDNA was ligated into prokaryotic expression vector pQE-80L and induced by IPTG to express HMGB1. The protein was purified with Ni~(2+)-NTA chromatography and polymyxin B affinity column. To identify the function of purified protein, the product was co-cultured with THP1 cells. Results Recombinant expression plasmid pQE-80L/HMGB1 was constructed successfully. After purification, the protein purity reached 96%. The recombinant HMGB1 stimulated THP1 to secrete TNF-? . Conclusion The highly purified HMGB1 was obtained successfully, which showed biological activity. These results lay the foundation for further research on the function of human HMGB1.

Objective To assess CT, MR manifestations and diagnostic value in hepatic tuberculosis. Methods CT findings in 12 cases and MR findings in 4 cases with hepatic tuberculosis proved by surgery or biopsy were retrospectively analyzed. Results (1) CT findings: one case of serohepatic type of hepatic tuberculosis was multiple nodular lesions in the subcapsule of liver. Parenchymal type were in 10 cases, including multiple,miliary, micronodular and low density lesions with miliary calcifications in 2 cases, singular, low density mass with multiple flecked calcifications in 3 cases, multiple cystic lesions in 1 case, multiple micronodular and low density lesions fusing into multiloculated cystic mass or “cluster” sign in 3 cases, and singular, macronodular and low density lesion with multiple miliary calcifications in 1 case. One case of tuberculous cholangitis showed marked dilated intrahepatic duct with multiple flecked calcifications in the porta hepatis. (2) MR findings in 4 cases were hypointense on both T 1WI and T 2WI in one case, hypointense on T 1WI and hyperintense on T 2WI in 3 cases. Enhanced MR in 3 cases showed slightly peripheral enhancement or with multilocular enhancement. Conclusions Various types of hepatic tuberculosis had their different imaging findings. The typical CT and MR findings could suggest the diagnosis.

Objective To assess the imaging and clinical manifestations of gastrointestinal mucosa associated lymphoid tissue(MALT)lymphoma with review of literatures. Methods Six cases of gastrointestinal MALT lymphoma (4 in stomach and 2 in intestine) proved by surgery and biopsy were examined with gastrointestinal barium meal examination (GI), CT or MRI scans. Results Gastrointestinal MALT lymphoma was an indolent disease that patients had a slow natural course, a high response rate to treatment and a long survival. Most cases of gastrointestinal MALT lymphoma had associated with Helicobacter pylori. GI showed thickened folds, multiple mucosal nodularity or polypoid filling defects, multiple point ulcers, and enlarged areae gastricae. CT and MRI revealed diffuse gastrointestinal wall thickening, polypoid lesion, and abdominal lymphadenopathy.Two or more findings and multiplicity of lesions might be the most important imaging features. Conclusions Based on the typical imaging findings combined with clinical characteristics, gastrointestinal MALT lymphoma could be suggested.

Objective　To assess the CT manifestations and diagnostic value in the pancreatic tuberculosis(PTB)with review of the literatures. Methods　All cases of PTB proved by surgery or biopsy were examined with plain and enhanced CT scans. Results　The CT findings in one case with multiple-nodular type of PTB were diffuse enlargement of the pancreas with multiple, nodular, and low-density lesions; The nodular lesions had peripheral enhancement. 7 cases of local type of PTB encroached on pancreatic head. 4 cases showed local soft tissue masses with multiple flecked calcifications in 2 cases and mild enhancement in one case; Cystic masses was found in 2 cases, with mural calcification in 1 case and multiloculated cystic mass in 1 case, respectively; Massive pancreatic head calcification was demonstrated in one case. In these 8 cases of PTB, the lesion extended out of pancreas in 4 cases, including abdominal tuberculous lymph nodes, tuberculous peritonitis, and hepatosplenic tuberculosis. Conclusion　CT findings of PTB were various but had some characteristics. Pancreatic masses with multiple flecked calcification or mild enhancement could suggest the diagnosis. Abdominal tuberculosis accompanied with the pancreatic lesion, especially tuberculous lymph nodes, was highly suggestive of the diagnosis of PTB.