Objective To screen proteins in pancreas cDNA library interacting with HBsAg protein.Methods After being amplified and identified,the human pancreas cDNA library plasmid was transformed into yeast Y187.Thereafter,the bait plasmid(PGBKT7-HBsAg) was transformed into yeast cells AH109.Then,two different transformed yeast cells were mated.The diploid yeast cells were plated on synthetic dropout nutrient medium(SD/-Trp/-Leu/-His/-Ade) and that medium contained X-?-gal for selection two times.After plasmids of true positive blue colonies were extracted and transformed to E.coli,the results were analyzed by DNA sequencing and blast searching in GenBank database.Results Six proteins binding to HBsAg were screened,including aconitase(ACO1),CDK5RAP3,carboxypeptidase,etc.Conclusion Most of these proteins play significant roles in the evolution of 2 type diabetes mellitus,which may supply some new clues for exploring the pathogenesis of abnormality of lipid metabolism and tglycometabolism in the patients with chronic hepatitis virus infection.

Objective To evaluate the efficacy of combined various surgical procedures for the treatment of post-traumatic epilepsy.Methods According to the preoperative primary location of epileptogenic focus and the monitoring results of intraoperative electrocorticography and depth electrodes,21 seizure patients were treated with various surgical procedures: 7 cases with epileptogenic focus resection,8 cases with epileptogenic focus resection combined with multiple subpial transection(MST),4 cases with anterior temporal lobectomy combined with amygdalo-hippocampotomy,1 case with selective amygdalo-hippocampotomy under neuronavigation,and 1 case with anterior callosotomy combined with MST and bipolar coagulation on funtional cortexes. Results All the patients were followed up from 3 months to 10 years postoperatively,and the surgical outcomes were such as the followings: satisfactory outcome in 9 cases(42.9%),marked improvement in 7 cases(33.3%),improvement in 3 cases(14.3%),slight improvement in 1 case(4.8%) and no improvement in 1case(4.8%).There was no died cases caused by the surgical procedures.Eight patients had temporary dysfunction after operation,but all recovered in four weeks after surgery.Conclusion The combined various surgical procedures are effective and safe for the treatment of post-traumatic epilepsy.

Objective To construct recombinant full length genotype B and C hepatitis B virus (HBV)and to examine HBV DNA replication and hepatitis B surface antigen(HBsAg),hepatitis B e antigen(HBeAg)expressions in Huh7 cells. Methods The full length genotype B and C HBV DNA were extracted and amplified from two HBV infected patients. The recombinant plasmids were constructed by inserting the amplified HBV fragments into the eukaryotic expression vector,pHY106,which were then transfected into Huh7 cells. The cells transfected with blank pHY106 vector were used as control. HBV DNA replication at 72 hours of transfection was detected by Southern blot. The HBV DNA levels in Huh7 cells at 24,48,72,96 and 120 hours of transfection were determined by real-time polymerase chain reaction(PCR).Meanwhile, the HBsAg and HBeAg expression levels in the supernatants at 24,48,72,96 and 120 hours were determined by enzyme linked immunosorbent assay(ELISA).Results The recombinant plasmids expressing genotype B or C HBV DNA were successfully constructed.The HBV replicative intermediates in HBV core particles,including rcDNA dsDNA and ssDNA,were detected by Southern blot.HBV DNA level could reach 8 lg copy/mL which was by real-time PCR. HBsAg and HBeAg levels determined by ELISA peaked at 72 hours after transfection and then declined gradually. Conclusions The recombinant plasmids inserted with genotype B or C HBV DNA are constructed successfully, which can express high levels of HBsAg and HBeAg in Huh7 cells. This system provides a platform for studying the pathogenesis of B and C genotype HBV, the interaction between HBV and host, as well as exploiting new drugs against HBV.

Objective To explore the clinical method and effect of ultrasonic osteotomy in mandibular angle reduction.Methods Under intronasal approach and general anesthesia,30 patients (5 male patients and 25 female patients) underwent intraoral prominent mandibular angle reductions with ultrasonic osteotomy.The jaw bone under periosteum was explored and the mental nerves protected.According to the clinical features of mandibular angle,curved mandibular angle osteotomy,mandible margin osteotomy or outer cotex splitting osteotomy were performed.Results All the osteotomy of prominent mandibular angle reductions could be completed with ultrasonic osteotomy.There were no complications of bleeding,nerve injury and malfracture happened.After 1 year following-up,the outlines of lower mandible were natural and concordant.Compared with traditional osteotomy devices,ultrasonic osteotomy was smoother and more precisely.Conclusions Ultrasonic osteotomy can improve the precise and safty of mandibular angle reduction.but compared with traditional osteotomy devices,it takes more time for osteotomy procedure.Surgeons need adapt to the different feeling and control methods.

Objective To screen proteins from human pancreas cDNA library,which interact with hepatitis C virus(HCV)E1 protein.Methods The human pancreas cDNA library was amplified,purified and evaluated,and then the purified library plasmids were transformed into yeast strain Y187.The reconstructed plasmid pGBKT7-E1 was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp.The transformed AH109 mated with Y187 that contained the library plasmids.The diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-α-gal for selecting.The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5α.The plasmids in DH5α were extracted,sequenced and blasted.Result Sixteen proteins interacting with HCV E1 were found.Conclusion Some of the sixteen pancreatic proteins may be related with metabolisms of glucose and lipid.

ObjectiveTo explore the changes in gene expression in tissues of patients with gastric carcinoma by cDNA microarray.MethodsA 44,000 gene cDNA microarray (Affymetfix) was used to examine gene expression in the tissues of patients with gastric carcinoma.The Sirt1 genes were identified and subjected to realtime PCR analysis.Results The expression of Sirt1 in tissues of gastric carcinoma [(0.318 ±0.032) ～ (0.169 ±0.013 ) time] was extremely lower than in tissues of para- gastric carcinoma ( 1 time).The results of Real-time PCR were consistent with those of genechip analysis.ConclusionsThe expression of Sirt1 was obviously different between tissues of gastric carcinoma and para - gastric carcinoma.Sirt1 may be associated with carcinogenesis of HCC.

group pACT-apoA Ⅰ and pBIND-core was higher than that in the negative control.Conclusion HCV core protein and apoA Ⅰ can interact in vivo.This study provides new theory for the mechanism of chronicity hepatitis C in fatty liver.

Objective To observe the operative effect of selective decompressive laminectomy and long segment pedical screw internal fixation for degenerative scohosis beyond 3 segments. Methods During January 2003 to July 2009, 52 cases with degenerative scoliosis beyond 3 segments were treated by posterior selective decompressive laminectomy, intervertebral implanted bone fusion and long segment pedical screw internal fixation, vetebral plate fusion seperately, and all the cases were followed up. Results All the patients were followed up for 12-60 months after operation. The status of back leg pain relief were evaluated through Oswestry score system. The Oswestry score of 1 week,3 months and 12 months after operation were (25.91 ± 9.05), (26.75 ± 8.37), (26.90 ± 8.45) scores respectively and there was statistical significance compared to the preoperative [(46.35± 8.12) scorns] (P ＜0.05). The average coronal Cobb angle was corrected from 15.23°± 3.35° to 10.02°± 3.26° 1 week after operation (P ＜ 0.05). All the internal implant didn't appear loosing or fracture confirmed by X-ray at 3,6,12 months after operation. The intervertebral implanted bone fusion and vetebral plate fusion all got well in 18-24 months after operation. Conclusion Selective decompressive laminectomy and long segment pedical screw internal fixation for the degenerative scoliosis can get better operative effect.

OBJECTIVETo observe the change in IL-13 mRNA expression in rat lungs with acute lung injury (ALI) induced by lipopolysaccharide (LPS), so as to investigate the significance of anti-inflammatory mechanism in the development of ALI.

METHODS(1) Different doses (2 approximately 8 mg/kg) of LPS were injected via the tail vein to the rats to establish ALI model. (2) The IL-13 mRNA content in pulmonary tissue was determined by RT-PCR.

RESULTS(1) Different degrees of ALI was induced in the rats by different doses of LPS. ARDS was produced in the rats by LPS of 6 mg/kg. (2) The IL-13 mRNA expression in the pulmonary tissue could be enhanced by the induction of LPS, especially when its dose was larger than 6 mg/kg.

CONCLUSION(1) Rat ALI model could be produced by LPS injection via the tail vein. (2) LPS in dose of 6 mg/kg was the threshold for the production of rat ARDS. (3) The incidence of rat ARDS might be closely related to the enhanced expression of IL-13 mRNA in rat pulmonary tissue.

Acute Disease ; Animals ; Disease Models, Animal ; Female ; Interleukin-13 ; biosynthesis ; genetics ; Lipopolysaccharides ; Male ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Respiratory Function Tests ; Respiratory Insufficiency ; chemically induced ; metabolism ; physiopathology

Objective:To evaluate the effect of electroacupuncture preconditioning on cellular immune function in the patients undergoing thyroid surgery.Methods:Sixty-six patients of either sex, aged 30-55 yr, with body mass index of 18.5-23.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification ⅠorⅡ, were selected and divided into electroacupuncture preconditioning group (group D) and control group (group C) by a random number table method, with 33 cases in each group. Patients in both groups received combined intravenous-inhalational anesthesia. Before anesthesia induction, both Hegu and Neiguan acupoints were stimulated for 30 min using a continuous wave at a frequency of 2/100 Hz, and the stimulation intensity was subjected to patient tolerance in group D. Group C received sham electroacupuncture for 30 min without stimulation. Venous blood samples were collected at 30 min before anesthesia induction (T 0) and 12, 24 and 48 h after anesthesia (T 1-3). The percentages of T lymphocyte subsets (CD3 +, CD4 +, CD8 +, CD4 + /CD8 + ) and natural killer cells and concentrations of interleukin-6 and tumor necrosis factor-alpha in the serum were measured by flow cytometry. Results:Compared with group C, the percentage of CD3 +, CD4 + and natural killer cells and ratio of CD4 + /CD8 + were significantly increased at T 1, 2, and the concentrations of interleukin-6 and tumor necrosis factor-alpha in the serum were decreased at T 1-3 in group D ( P<0.05). Conclusions:Electroacupuncture preconditioning can improve cellular immune function in the patients undergoing thyroid surgery.