Objective To obtain abundant human betacellulin(BTC) with biological activity. Methods The whole mature protein coding sequence of BTC gene was amplified by polymerase chain reaction(PCR) method applied to human pancreatic ?-cell tumors cDNA.The fragment was cloned into prokaryotic expression vector pET32a(+) plasmid.The recombinant plasmid was transformed into E.coli BL21 and the fusion protein was expressed under isopropyl-beta-D-thiogalactopyranoside(IPTG).The fusion protein was purified by Ni2+ affinity chromatography.SDS-PAGE and Western blot were employed to determine the expression and purification of the expected protein.BTC was added to culture NIH3T3 cells for 5 days,and cell proliferation was detected by MTT. Results Lots of fusion protein were produced,and the purified protein can stimulate the proliferation of NIH3T3 cells. Conclusion The human BTC can be successfully obtained from the pET32a(+) system with the biological activity of stimulating the proliferation of NIH3T3 cells.

Objective To construct eukaryotic expression vector of human pancreatic duodenal homeobox 1(PDX-1) gene,and to detect its expression in NIH3T3 cell lines. Methods The whole coding sequence of PDX-1 gene was amplified by polymerase chain reaction(PCR) from human pancreatic-cell tumors cDNA.The fragment was inserted into eukaryotic expression vector pcDNA3.1 plasmid.The recombinant plasmid was verified by double digestion and DNA sequencing.The expression of PDX-1 gene in NIH3T3 cells was assayed by Western blot. Results The length of specific fragment amplified by PCR was 852 bp,and the recombinant plasmid pcDNA3.1-PDX-1 showed two bands of 5.5 kb and 852 bp by digestion using respective restriction enzymes BamHⅠand EcoRⅠ.The sequence of PDX-1 gene was approved or confirmed by blasting to GenBank.It was suggested that PDX-1 gene had been cloned into pcDNA3.1 vector correctly.Western blot showed that PDX-1 gene was expressed,which was detected 24 h after pcDNA3.1-PDX-1 plasmid was transfected into NIH3T3 cells. Conclusion The recombinant eukaryotic expression vector pcDNA3.1-PDX-1 was successfully constructed and expressed in NIH3T3 cell lines.

Objective To investigate the efficacy and safety of laparoscopic operations of ovarian tumors in children.Methods A total of 24 children with ovarian tumors treated laparoscopically from January 2000 to March 2005 were studied.The laparoscopic operations(tumor enucleation or unilateral adnexectomy) were conducted under general anesthesia with endotracheal intubation.Electrocoagulation of the capsule was performed alongside the borderline of the lesion for tumors 5 cm in diameter.After blunt dissection of the tumor and capsule,the lesion was completely enucleated.Hemostasis of the residual cut surface was accomplished by using electrocoagulation,with no need of suture. For large ovarian cysts disturbing laparoscopic visualization,the aspiration of hydatid fluid was conducted before the enucleation.Results All the operations were successful completed under laparoscope,including 24 cases of tumor enucleation and 2 cases of unilateral adnexectomy.The operation time was 50?20 min,the amount of hemorrhage was 30?10 ml,and the postoperative hospital stay was 2~4 d.No surgical complications or wound infection occurred.Follow-up in the 24 cases for 1 year found no recurrence.Conclusions Laparoscopic operations for ovarian tumor in children is safe and effective.

Health Policy ; Humans ; Italy ; Occupational Health Services ; organization & administration ; Safety Management

Finland ; Health Policy ; Humans ; Occupational Health Services ; organization & administration ; Safety Management

Objective To facilitate standing and walking after rehabilitation training by designing and using individualized ischial weight bearing orthosis for patients with various conditions of the lower limbs. Methods An orthosis with ischial support was ordered and tailored according to the condition of the patient's affected lower limb. Adaptive standing and walking training were administered daily. The functional recovery after the assembly of the orthosis was observed. Results The patients' standing and walking ability improved significantly after the training with the ischial weight bearing orthosis. The standing time was significantly longer than without the orthosis, and the walking distance was notably increased. Conclusion An ischial weight bearing orthosis has a significant effect on the recovery of standing and walking capability.

Objective To investigate the feasibility and safety of transumbilical laparoscopic single-port choledocholithotomy in the treatment of common bile duct stones. Methods The clinical data of four patients who underwent transumbilical laparoscopic single-port choledocholithotomy at Shengjing Hospital of China Medical University from January to June, 2010 were retrospectively analyzed. The operation method and postoperative effects were reviewed. Results All the operations were successfully carried out. The mean operation time was 100 minutes and the mean blood loss was 62. 5 ml. All patients were cured without the occurrence ofcomplications except for one patient who had a peritoneal infection and was cured after surgical drainage. Conclusion Transumbilical laparoscopic single-port choledocholithotomy is safe and feasible, but indications should be strictly followed.

Objectives To evaluate the effect of N-Acetylcysteine on the airway inflammation and remodeling of obese asthma mice with high-fat diets. Methods Forty female C57BL/6J mice were randomly divided into 4 groups, asthma group (A), obese asthma group (B), treatment group (C) and normal control group (D). Group A were sensitized and challenged by ovalbum (OVA) and normal diets. Group B were sensitized and challenged as group A but fed with high-fat diets, while group C were sensitized, challenged and fed as group B, but administrated N-Acetylcysteine 200 mg/kg. d from the third week after challenge. The cells in BALF were counted and classified after staining, IL-6 and 8-iso-PGF2α(8-iso-PGF2α) in lung tissues were detected by ELISA. WAt, WAm, Pbm, as the remodeling indices, measured in lung pathological section. All parameters were compared among 4 groups. Results In comparison with group D, the leukocytes and EOS in BALF, WAt/Pbm, WAm/Pbm in lung section were increased as well as IL-6 and 8-iso-PGF2α in lung tissue elevated in group A and group B, while the maximum changes were observed in group B (P ＜0. 05). After NAC treatment, the IL-6, 8-iso-PGF2α and WAt/ Pbm were decreased significantly (P ＜0. 05). Pearson correlation analysis showed that WAt/Pbm and IL-6 were in positive correlation with 8-iso-PGF2α (r =0. 817, 0. 737, P ＜0. 01). Conclusions N-Acetylcysteine can alleviate the airway inflammation and remodel reaction of asthma by a substantial inhibition of the oxidative stress reaction in lung.

Objective To investigate the epidemiological characteristics and risk factors of clonorchiasis of hospital patients in Heilongjiang Province,so as to provide the evidence for formulating the control and prevention strategies. Methods A total of 2 359 suspected patients from human parasitic disease research institute of Harbin medical university were investigated. Total?ly 6 718 stool samples and 2 359 serum samples were tested by Kato?Katz technique and the enzyme linked immune method re?spectively. Meanwhile,the information of the patients was collected by questionnaires. Results Totally 513 suspected patients were infected with Clonorchis sinensis. The infection rate of the suspected patients was higher in the ≥29 group(P<0.05), and the infection rate and positive rate of blood tests of the rural suspected patients were both higher than those of the urban sus?pected patients(both P<0.05). The habit of eating raw fish and shrimp was the risk factor of clonorchiasis. Conclusion Clo?norchiasis is one of the main food?borne parasitic diseases in Heilongjiang Province. The habit of eating raw fish and shrimp is the risk factor of clonorchiasis.

Objective To explore the establishment methods of animal models of adult growth hormone deficiency, and to provide a good model for experimental research and treatment for abnormal bone metabolism caused by growth hor-mone deficiency.Methods The methods of establishment of animal models of adult growth hormone deficiency were re-viewed and evaluated refering to literature.Results There were three methods including spontaneous lack-of, pituitecto-mized and gene knockout can establish animal models of adult growth hormone deficiency.Conclusions Hypophysecto-mized animal models are inexpensive and easy to create, but not suitable for studying the relationship between growth hor-mone and bone metabolism.Spontaneous lack-of and gene knockout models are specific growth hormone deficiency and of great research significance in exploring the relationship between growth hormone and bone metabolism.