Abstract Objective: To construct the Second half of CILP(C2) MBP fusion protein by sub-cloning technology. Methods: Recombinantfusion proteins, which contain the fragments within the C2 region(designated C2F1, C2F2 and C2F3) of the non-porcine nucleotide pyrophos-phohydrolase-homologous region of CILP, were prepared using pMAL-eHis vector. The recombinat genes are induced by different temperatures(22℃,30℃,37℃ ). Results: Expression using pMAL-eHis system can be induced chemically by adding IPTG. 37℃ temperature prmotes in-soluble inclusion-body formation,but 22℃ temperature can not induce the enough expression of recombinant protein. Onl 30℃ temperaturecan induce enough amount of soluble recombinant protein. The characers of fusion proteins that they carried 6 straight histidines, (His)6, at tbeC-terminus of multiple cloning sites for affinity purification were assessed by sodium dodecy1 sulfate-polyacylamide gel electrophoresis(SDS-PAGE) and Western blot. Nucleotide sequences of the insertion genes were confirmed by dideoxy sequencing. Conclusion: C2F1, C2F2,C2F3-MBP fnsion proteins were constructed successfully.These recombinant proteins may provide important roles in the future study on CILP.

To compare the anxiolytic effects of reduced metabolite of progesterone and benzodiazepine.Methods　The effects of allopregnanolone and diazepam on spontaneous locomotor activity and on exploration in the elevated plus-maze were studied in C57 mice 20 min after vehicle or drug intraperitoneal administration.Results Allopregnanolone (0.1 mg.kg-1,ip) elicited marked anxiolytic effects in terms of significantly reducing the latency to enter the open arm from (31.30±8.39)s to (8.80±6.00)s,(P<0.001),and significantly increasing both the number of open arm entries from 1.20 ± 0.42 to 4.80 ±1.75,(P<0.001) and the proportion of total time spent on the open arm from 7.13％ to 32.50％,(P<0.001).Meanwhile,the diazepam (0.25 mg·kg-1) produced a lower anxiolytic effect comparing to that of the allopregnanolone.Analysis of spontaneous locomotor activity showed while 0.5 mg·kg-1 of diazepam decreased the locomotor activity (P<0.01),neither 0.1 mg·kg-1 of allopregnanolone nor 0.25 mg·kg-1 of diazepam affect the locomotor activity score.Conclusion Together,these results provide evidence for differential behavioral actions of the neurosteroids and benzodiazepines.Since the allopregnanolone produce a selective anxiolytic effect without affecting the spontaneous locomotor activity,the allopregnanolone may be a better alternative for diazepam in treating anxiety.

Objective To analyze the clinical effect of debridement in the treatment of Kashin-Beck disease under an ankle arthroscopy.Methods Totally 40 patients with ankle Kashin-Beck disease who had underwent surgery in the Fifth Hospital of Daqing from January 2015 to October 2016 were selected as the research subjects.All patients were treated with cleaning treatment under arthroscopy,evaluated via the visual analogue scale method,and compared pain before and 6,12 month after the treatment.Walking function and ankle motion of the patients were evaluated via an ankle-scoring system,and the quality of life of patients was investigated before and after the treatment through a questionnaire survey.Results The visual analogue scale of before and 6,12 months after the treatment was (6.2 ± 1.0),(2.3 ± 1.2) and (1.6 ± 0.4) scores,respectively;ankle foot score (53.3 ± 6.8),(86.6 ± 5.1) and (88.7 ± 6.2) scores,respectively;ankle activity score (15.9 ± 4.1),(36.9 ± 4.9) and (41.8 ± 6.4) scores,respectively;and life quality score (53.1 ± 4.0),(75.4 ± 6.6) and (88.7 ± 10.5) scores,respectively.The differences between groups were statistically significant (F =2.1,2.6,2.3,2.5,P ＜ 0.05).Visual analogue scale of 6 and 12 months after the treatment was significantly lower than that before the treatment,ankle foot score and ankle activity score and life quality score were significantly higher than those before the treatment (P ＜ 0.05).Conclusion The debridement under an ankle arthroscopy can effectively reduce the pain of patients with Kashin-Beck disease,and improve the walking function of the patients obviously,significantly improve the patients' life quality,and the method should be widely applied in clinical treatment.

AIM: To study the effect of genistein on c-myc mRNA expression induced by oxidized low density lipoprotein (ox-LDL) in human vascular endothelial cells (ECV304). METHODS: LDL were isolated from healthy human plasma by gradient ultracentrifugation and oxidized by CuSO4. ECV304 cells were exposed to ox-LDL 200 mg*L-1 in the presence or absence of genistein 100 μmol*L-1 for 1, 2, and 4 h in vitro. Northern blot was employed to measure c-myc mRNA levels of ECV304. RESULTS: In response to ox-LDL 200 mg*L-1, c-myc mRNA expression in ECV304 increased by 3 fold for 1h and 3.3 fold for 2 h and decreased below the control level at 4 h. Expressions of c-myc stimulated by ox-LDL in the presence of genistein 100 μmol*L-1 for 1 h and 2 h were separately 80 percent and 60 percent of that in the absence of genistein 100 μmol*L-1. CONCLUSION: Genistein can effectively inhibit c-myc mRNA expression in ECV304 induced by ox-LDL.

Objective To investigate the efficacy of autologous peripheral hematopoietic stem cell transplantation in the treatment of adult dermatomyositis. Methods A 21-year-old patient with dermatomyositis received autologous peripheral hematopoietic stem cell transplantation and was followed up for 6 years. Autologous peripheral hematopoietic stem cells were mobilized by recombinant human granulocyte colony stimulating factor (rhG-CSF) before the transplantation, and the conditioning regimens consisted of cyclophosphamide,methylprednisolone and cyclosporin. Rabbit anti-human T lymphocyte immunoglobulin began to be applied on day 3 after retransfer of stem cells. The improvement in symptoms, physical signs and biochemical indicators was observed, and hematopoietic restructuration and immunity resurrection were evaluated after the transplantation. Results After the transplantation, skin eruption greatly improved and gradually subsided. The muscle force of extremities restored from level Ⅳ before transplantation to level Ⅴ. The level of creatine kinase declined sharply after transplantation, but gradually returned to previous level. Leucocyte count began to decrease on the day of retransfer, and returned to the normal level on day 8. Immune function remained normal before and after the transplantation. Conclusion Autologous peripheral hematopoietic stem cell transplantation is an alternative treatment for severe and refractory dermatomyositis.

Animals ; Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Hydrogen-Ion Concentration ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Oxygen ; metabolism ; Rats

BACKGROUND:Coracoclavicular ligament reconstruction with transclavicular-transcoracoid driling is an effective surgical technique to treat acromioclavicular dislocation. A good driling in the clavicle leads to a perfect bony tunnel and a good surgery. OBJECTIVE: To observe the effects of different driling positions of the clavicle on the location of bony tunnels in coracoclavicular ligament reconstruction. METHODS:Sixty three-dimensional digital models of the clavicle and coracoid process were constructed by Mimics13.0. Virtual transclavicular-transcoracoid bony tunnels were established according to different surgical planes with different driling positions in the clavicle. Parameters of these bony tunnels were measured, and the safety was evaluated. Option 1: The driling was made 30 mm distal to the clavicle, located in the center of the front and rear edges of the clavicle surface. Option 2: The driling was made 40 mm distal to the clavicle, located in the center of the front and rear edges of the clavicle surface. Option 3: The driling was made at the straight line of tapered nodule tip and the midpoint of the base of the coracoid process, located at the rear edge of the clavicle upper surface. RESULTS AND CONCLUSION: Bony tunnels in option 1 were extremely on the inside of the coracoid. Bony tunnels in options 1 and 2 were not in the center of clavicle. Bony tunnels in option 3 were in the center of both clavicle and coracoid. The method of locating the driling position with a certain distance to the distal clavicle leads to different results in man’s and woman’s models. To ensure that the bony tunnel can pass through the center of clavicle and coracoid, it is suggested to dril at the straight line of tapered nodule tip and the midpoint of the base of the coracoid process and nearby the rear edge of the clavicle upper surface.

Object To investigate the application of cell immobilization culture to producing ginkgolides in Ginkgo biloba L. Methods To immobilize the cells by polyaminoresin foam as support materials, and determine the effects of different factors. Results The results showed that 71% of immobilization ratio and 22 mg/cm 3 of cell density were obtained using P29 of high density, small bore diameter with pieces of 0.5 cm ? 0.5 cm ? 0.5 cm, 0.72 g of support matrices per bottle in 65 mL of MS medium solution supplemented with 2,4-D 8.0 mg/L+KT 0.04 mg/L+NAA 0.4 mg/L and the incoulum quantity was 200 g/L. Conclusion It shows that present method has some advantages, such as simpleness, low cost, high immobilization ratio.

Aim To compare the anxiolytic effects of reduced metabolite of progesterone and benzodiazepine.Methods The effects of allopregnanolone and diazepam on spontaneous locomotor activity and on exploration in the elevated plus_maze were studied in C57 mice 20 min after vehicle or drug intraperitoneal administration. Results Allopregnanolone (0.1 mg?kg-1, ip) elicited marked anxiolytic effects in terms of significantly reducing the latency to enter the open arm from (31.30?8.39)s to (8.80?6.00)s, (P