Objective:To clone the mice interleukin-23(mIL-23) and express it in Pichia efficiently.Methods:Two subunits of IL-23 were amplified by PCR from pcDNA3/mIL-23,and ligated together with adaptor by over-PCR.The IL-23 cDNA confirmed by sequencing was inserted into expressing vector pPICZαA and expressed in Pichia X-33 strain.IL-23 protein expression was induced by methanol and ammonia.The recombinant IL-23 was identified by immunoblot and its biological activity was analyzed.Results:DNA sequencing confirmed that cloned cDNA was identical to the published sequence of mIL-23.The recombinant plasmid pPICZαA/mIL-23 was electroprated into X-33.An expected 72 kD protein of mIL-23 was founded both in the induced pellets and supermatants by SDS-PAGE and coomassie blue staining.The 72 kD protein could be recognized in Western blot.While we could detect bands at 72 kD in cell pellets induced more than 24 h by Western blot.Detected by Western blot using anti-His antibody,we could see positive band at 72 kD from supernatants induced 24 and 36 h.While we could detect band at 72 kD in cell pellets induced between 24 h and 96 h.Meanwhile,we could see obviously proliferation of mice peripheral blood mononuclear cells(PBMC) treated with the induced pellets and supermanants.Conclusion:We have successfully expressed mIL-23 protein in Pichia and the expressed product has its bioactivity in promoting the proliferation on PBMC.

OBJECTIVETo detect the expression level of interleukin 17 (IL-17) in the plasma and colonic mucosa of patients with ulcerative colitis (UC), and to explore the synergistic mechanism of qingchang huashi recipe (QHR) combined with Mesalazine.

METHODSRecruited were 24 mild or moderate UC patients of damp-heat inner accumulation syndrome (DHIAS). Their samples of intestinal tissues were histologically graded. They were assigned to the combination group and the Western medicine (WM) group, 12 in each group. Besides, another 12 healthy volunteers were recruited as the healthy control group. QHR combined Mesalazine were given to patients in the combination group, while those in the WM group took Mesalazine. The therapeutic course for all was 3 months. By the end of treatment the expression level of IL-17 in the plasma and colonic mucosa was detected using ELISA. The infiltration of IL-17 in the intestinal mucosal tissue was detected by immunohistochemical SP method.

RESULTSThe expression level of IL-17 in the plasma and colonic mucosa was significantly higher in UC patients than in healthy controls (P <0. 05). The higher the histological grading the higher the expression level. The expression level of IL-17 in plasma and colonic tissues decreased after treatment in the two treatment groups (P < 0.05). Besides, the expression level of IL-17 was lower in the combination group than in the WM group (P <0.05).

CONCLUSIONQHR combined Mesalazine could synergically enhance the effect and effectively inhibit intestinal inflammation through down-regulating the expression of IL-17.

Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Colitis, Ulcerative ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Immunologic Factors ; metabolism ; Inflammation ; metabolism ; Interleukin-17 ; metabolism ; Intestinal Mucosa ; drug effects ; Intestines ; metabolism ; Mesalamine ; therapeutic use

BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results of BMSCs/ HAB composite. RESULTS: ① Iliac bone block cells were cleaned with good reservation of bone matrix. ② The level of ALP and OCN of MSCs was higher after in ducing for 8 days than that in control group [MSCs after induction: (181.54±40.01) nkat/L, (7.2±1.3) μg/L. There was no method to detect the level in control group, P ＜ 0.05]. ③ BMSCs were adhered and grew well in HAB scaffold. CONCLUSION: HAB loaded with induced BMSCs has an excellent os teogenic function in vitro and shows an effective potential as a good bone tissue engineering material.

Aptamers are capable of binding a wide range of biomolecular targets with high affinity and specificity. It has been widely developed for diagnostic and therapeutic purposes. Because of unique three dimensional structures and cell-membrane penetration, aptamers inhibit virus infection not only through binding specific target, such as the viral envelope, genomic site, enzyme, or other viral components, but also can be connected to each other or with siRNA jointly achieve antiviral activity. Taking human immunodeficiency virus and hepatitis C virus as examples, this paper reviewed the effects and mechanisms of aptamers on disturbing viral infection and replication steps. It may provide an insight to the development of aptamer-based new antiviral drugs.

Ten compounds were isolated from the 70% ethanol extract of linseed meal (Linum usitatissimum L) through a combination of various chromatographic techniques, including silica gel, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as 1-methylethyl-2-O-beta-D-glucopyranosyl-(1" --> 6')-beta-D-glucopyanoside (1), linustatin (2), neolinustatin (3), lotaustralin (4), linamarin (5), deoxyguanosine (6), deoxyadenosine (7), (+)-pinoresinol-4'-O-beta-D-glucopyranoside (8), 4-O-beta-D-glucopyranosylvanillyl alcohol (9) and tachioside (10), separately. Among them, compound 1 is a new compound, and compounds 6, 8 and 10 were isolated from the linseed meal for the first time.

BACKGROUND: Zein has excel ent solubility, heat resistance and biodegradability, but its biocompatibility and effect on periodontal defects repair are under discussion. OBJECTIVE: To analyze biocompatibility of zein and its effect on periodontal defect repair. METHODS: Zein scaffold was prepared by solvent casting/particulate leaching. In vitro test: Human periodontal ligament cel s were co-cultured with zein scaffold for 18 days, and cel growth was observed by scanning electron microscope. In vivo test: Eight Beagle dogs were enrol ed to establish periodontal defect models, which were randomly assigned to receive zein scaffold implantation as experimental group, or interrupted suture as control group. Afterwards, the defect region was observed by scanning electron microscope at 3 months. RESULTS AND CONCLUSION: In vitro results: Human periodontal ligament cel s adhered wel and tightly on the scaffold with a fusiform, and could grow around pores. In vivo results: In the experimental group the scaffold dissolved completely, bone trabecular arranged regularly, and mature tissues appeared, to be integrated with the surrounding tissues; in the control group, the defect region almost healed, but there were irregular fibers and obvious lacunae. Moreover, compared with the control group, the height of new alveolar bone and bone defect, as wel as the length of junctional epithelium were significantly decreased, and new cementum was significantly increased in the experimental group (P < 0.05). To conclude, zein scaffold has good biocompatibility and can promote periodontal defect repair.

OBJECTIVE To study the sterilization method for on-wall oxygen humidifying inhalation sets in hospital.METHODS The different parts of on-wall oxygen humidifying inhalation sets were sterilized with Jianzhishu disinfectant and Andofo povidone iodine disinfectant liquid.RESULTS The regular percents for bacterial examination of the different parts of oxygen humidifying inhalation sets after sterilization were higher than before,P

OBJECTIVE To study the sterilization effect of on-wall oxygen humidifying inhalation sets.METHODS The different parts on on-wall oxygen humidifying inhalation sets were sterilized,then compared the result before and after sterilization.RESULTS The regular percents for bacterial examination of humidifying bottle were 17.14% before sterilization,and 100.00% after sterilization,the regular percents for bacterial examination of metallic guilloche were 34.29% before sterilization and 94.29% after sterilization,the regular percents for bacterial examination of vent tube in humidifying bottle were 8.57% before sterilization,and 97.14% after sterilization.The regular percents for bacterial examination of the different parts of oxygen humidifying inhalation sets after sterilization were higher than before it P

Objective To evaluate the effect of applying emergency care simulator (ECS)on cardiopulmonary resuscitation skill training. Methods Clinical doctors and nurses(n=1 472)received the skill training of unarmed cardiopulmonary resuscitation (CPR),electric defibrillation and tracheal intubation by using simple CPR simulator (n=667)or ECS (n=805)respectively. After training,the examination was carried out and questionnaires were handed out. The completion time of operations, the scores and pass rates of examinations,the evaluations of training methods were compared between the two groups. The statistical data were analyzed by t(t')-test or Chi-square test with SPSS 19.0 and P <0.05 was considered as the criterion of significance. Results Compared with those of group using simple CPR simulator,the group using ECS showed obviously shortened completion time((309±125)s vs. (242±61)s,t'=12.65,P=0.00;(87±36)s vs. (55±31)s,t'=20.28,P=0.00;(239±87)s vs. (145±53),t'=24.4,P=0.00),significantly increased scores ((83.5±14.8)vs. (90.2±17.6), t'=7.93,P=0.00;(84.7±19.3)vs. (92.1±21.5),t'=6.95,P=0.00;(81.6±15.3)vs. (89.6±13.5), t'=10.53,P=0.00)and pass rates of examinations (84.1%(561/667)vs. 92.5%(745/805),χ2=25.96, P<0.01;82.2%(548/667)vs. 91.2%(734/805),χ2=26.41,P<0.01;80.8%(539/667)vs. 91.4%(736/805),χ2=35.48,P<0.01)as well as higher evaluation of training methods. Conclusions Application of ECS can obviously improve the training effect of CPR and it can be widely popularized in the edu-cation and training of emergency medicine skills.

2.83 cm(left-right).The density of lymphocele was homogeneous ,with 0~15 HU(mean value,5.7HU).Adjacent organs were displaced by compression of lymphocele.Conclusion It should be diagnosed as a lymphocele if a pelvic cyst is found in postoperation of pelvic malignant tumor.