By using gene SOEing PCR, Arg (CGC) in the expression fragment of chitinase gene (chi58) from Chaetomium cupreum was mutated synonymously to Arg (AGA), which is a bias code of Pichia pas-toris yeast. The expression plasmid pPIC9K-chi58A was constructed and transformed into GS115 strain through electroporation. The chitinase activity reached 101.71 U/mL?3.33 U/mL after induced with 120 h, which was the 3 fold of the original strain (31.83 U/mL?4.85 U/mL). Additionally, recombinant yeast showed good genetic stability after 10 cycle. SDS-PAGE analysis suggested that the weight of protein was 58 kD.

Objective:To produce a fusion protein(ICOS-Ig)between extracellular portion of human ICOS and Fc portion of mouse IgG 2a and study on its biological activity.Methods:cDNA encoding the extracellular domain of human ICOS was prepared by RT-PCR from the RNA of the stimulated human peripherial blood mononuclear cells. The Fc portion of mouse IgG2a was cloned by PCR from the vector that contains the sequence-encoding Fc portion of mouse IgG2a.Two resulting amplified PCR products were ligated into a secrection mammalian expression vector, pSecTag2/Hygro A. The recombined vector was transfected into CHO cells by lipofectamine2000.Results:RT-PCR demonstrated the integration and mRNA synthesis of fusion gene. ELISA and Western blot analysis showed the expression of ICOS-Ig and its molecular weight was between 43-66 kD, its concentration was 5-25 ?g/ml. FACS analysis assured that ICOS-Ig had ligand specific binding activity. Addition of ICOS-Ig to MLR resulted in inhibition of T-cell proliferation and IL-2,IFN-? secretion.Conclusion:The fused gene ICOS-Ig was constructed and expressed successfully. It had the bioactivity of inhibition of T cell proliferation and cytokine secretion.

Objective To explore the relationship between organochlorine pesticides exposure after Tangshan earthquake and the breast cancer. Methods 1∶1 matched case-control study was conducted in 150 newly diagnosed cases of breast cancer with histopathological diagnosis from three city-level hospitals in Tangshan,China and 150 matched controls at the same hospitals,without the related diseases. The levels of derivatives of DDT and HCH in serum were measured by gas chromatography (GC) method and epidemiological survey were conducted by questionnaire for 150 pairs of cases and controls. The relationship between the organochlorine pesticides metabolites and the breast cancer of the exposed population was analyzed by conditional logistic regression model by SAS 8.2 Software. Results Compared with the control,significant higher rates of detection of p,p'-DDD and o,p'-DDT were seen in the case group and the levels of p,p'-DDE,?-HCH and ?-HCH in serum were higher in the case group,and there was a significant difference in serum residues levels of ?-HCH,p,p'-DDE and o,p'-DDT in different exposure areas in Tangshan,the high exposure area showed higher residues levels than the low exposure area. Multivariate conditional logistic regression analysis showed high pesticides exposure after the earthquake,the serum residues of p,p'-DDE and p,p'-DDT were associated with breast cancer risk,ORs were 6.592(95%CI:1.402-31.002),2.405(95%CI:1.033-5.600)and 1.845(95%CI:1.028-3.353) respectively. Conclusion Exposure to environmental organochlorines may be one of the risk factors for breast cancer; Higher pesticides exposure after the earthquake may increases breast cancer risk in Tangshan area.

Objeetive To investigate the optimized concentration,effectiveness and histological change of Pingyangmycin by local injection of keloids for two weeks.Methods The appropriate concentration was analyzed,based on the Vancouver scar scale.48 eases of keloids were randomly divided into 4 groups with different concentrations of Pingyangmycin (1,0.5,0.25,and 0.125 mg/ml).Each case was iniected for five times at two weeks intervals,and then the statistical results (pair t test) and the histological changes were evaluated.Results Statistically,0.5 mg/ml group was more effective (P=0.0026),and this group had significant differences from the others (SNK test).The rate of the recurrence for one year was 12.00% in the whole effective cases.The keloid tissue after treatments,histologically,showed the thicker epidermis and larger papillary layer of the dermis,more inflammatory cells.Conclusions 0.5 mg/ml Pingyangmycin is the most appropriate concentration in this study and 0.25 mg/ml is the effective one.This treatment for keloid is convenient,safe,effective,low recrudescence and few side effects.

Objective To improve hand hygiene compliance among healthcare workers (HCWs)in a stomatological hos-pital,and effectively reduce healthcare-associated infection(HAI)rate.Methods According to WHO hand hygiene guide-line and implementation scheme of hand hygiene,a systemic and step by step hand hygiene promotion plan was formulated, the plan was divided into five steps:full alert,baseline investigation,improvement,tracking,and retrospective analysis. Hand hygiene compliance among HCWs before and after the promotion of activities was analyzed and compared. Results Through hand hygiene promotion activities,the questionnaires showed that average score of knowledge about hand hygiene among HCWs enhanced from 58.33 to 77.40 (increased by 32.69%);the average hand hygiene compliance rate increased from 24.28%(59/243)to 49.81%(133/267),the difference was significant(χ2 =35.331,P =0.000);hand hy-giene status at five moments for hand hygiene improved markedly.Conclusion Hand hygiene promotion has a significant in-fluence in hand hygiene among HCWs in this stomatological hospital.

Objective To investigate the effect of running exercise on cartilage in rats with an unstable knee joint.Methods Twenty 8-week-old Sprague-Dawley rats had their left anterior cruciate ligament cut to model an unstable knee.They were randomly divided into a control group and an experimental group,10 rats in each group.The control group was given no intervention,while the experimental group accepted running exercise training on na animal treadmill at a velocity of 15 m/min for an hour every day.After 3 and 6 weeks of training,5 rats were sacrificed and cartilage from the medial condyle of the femur was sampled,decalcificated,embedded and sliced on the sagittal plane.After hematoxylin-eosin staining,toluidine blue staining and immunohistochemical staining,the cartilage thickness,Mankin' score,the content of matrix collagen and the proteoglycan content of the cartilage matrix were assessed,and the shape and structure of the unstable knee joints were observed under a transmission electron microscope.Results The cartilage thicknesses and Mankin's scores at 6 weeks were significantly different from those at 3 weeks in both groups.In the experimental group the average thickness of cartilage was 154 ± 13 μm at 3 weeks and 131 ± 15 μm at 6 weeks.The corresponding Mankin's scores were 9.93 ± 1.36 and 11.23 ± 1.57,respectively.Both were significantly different from the control group averages at the same time points.There was also a significant difference in the positive rate of toluidine blue and collagen type Ⅱ staining between the experimental group and the control group at both time points,and in the experimental group between 3 and 6 weeks of training.After 3 weeks of training,fewer chondrocytes were observed under the transmission electron microscope in the experimental group,and fissures were seen on the surface of the cartilages.However,3 weeks later,quite a few ruptures and a lot of necrotic cells could be seen.Conclusions Running exercise can damage the cartilage of unstable knee joints and speed up the development of osteoarthritis.Even moderate exercise could aggravate damage to unstable joints and the cartilage matrix,and accelerate chondrocyte degeneration.

Objective To investigate the correlation between high myopia and MMP-2 in aqueous humor (AH), by comparing the protein level of MMP-2 in AH from patients with cataract combined with high myopia and with emmetropia. Design Experimental study. Participants 30 AH samples of patients with age-related cataract. Methods AH samples were collected from the patients with age-related cataract during phacoemulsification, including 15 AH samples from patients combined with high myopia (experimental group) and 15 AH samples from patients with emmetropia (control group). The expression of pro-MMP-2 and MMP-2 in AH were ana-lyzed by Western blot technique. Main Outcome Measures The gray value of MMP-2 protein band detected by Western blot. Results The level of pro MMP-2 was statistically higher than that of MMP-2 in experimental group (430.4±57.3 versus 294.5±35.2, t=10.400, P= 0.000) and control group (402.8±57.7 versus 280.3±49.7, t=8.400, P=0.000). There was no statistically difference in the level of pro-MMP-2(t=1.320, P=0.200)and MMP-2 (t=0.900,P=0.375) between the two groups. Conclusions No abnormal expression of MMP-2 was detected in AH from patients with high myopia, according to this study based on limited samples. Pro-MMP-2 was the main form of MMP-2 in AH of patients with cataract combined with high myopia or emmetropia, which possessed potential ability of transferring into form of active MMP-2. (Ophthalmol CHN, 2009, 18: 260-264)

Objective To evaluate the effect of dexmedetomidine on the quality of recovery from sevoflurane-based anesthesia in the patients undergoing abdominal surgery.Methods Eighty patients of both sexes, aged 30-64 yr, weighing 45-80 kg, with American Society of Anesthesiologists physical status Ⅰ or Ⅱ , scheduled for elective abdominal surgery, were randomly assigned into 2 groups (n =40 each) using a random number table: control group (group C) and dexmedetomidine group (group D).After routine induction of anesthesia, all the patients were tracheally intubated, and mechanically ventilated.Anesthesia was maintained with sevoflurane inhalation and intermittent iv boluses of sufentanil and cisatracurium besylate, and bispectral index values were maintained at 45-60.After induction of anesthesia, dexmedetomidine was infused in a dose of 1 μg/kg over 15 min, followed by a continuous infusion of 0.6 μg · kg-1 · h-1 until 30 min before the end of surgery in group D.Group C received normal saline instead.Heart rate (HR) and mean arterial pressure (MAP) were recorded at the end of surgery, and at 1 min before and after extubation.The emergence time, extubation time, and occurrence of bucking during extubation, and nausea and vomiting, respiratory depression, and agitation during recovery from anesthesia were recorded.Results Compared with group C, MAP and HR were significantly decreased at 1 min before and after extubation, the emergence time and extubation time were shortened, the incidence of bucking, nausea and vomiting, and agitation was decreased, and no significant change was found in the incidence of respiratory depression in group D.Conclusion Dexmedetomidine infused in a dose of 1 μg/kg after routine induction of anesthesia, followed by a continuous infusion of 0.6 μg · kg-1 · h-1 until 30 min before the end of surgery, can significantly increase the quality of recovery from sevoflurane-based anesthesia in the patients undergoing abdominal surgery.

OBJECTIVE: To explore the effects of Gymnadenia conopsea alcohol extract (GcAE) on the collagen synthesis in rat lungs exposed to silica and the influence on antioxidase activities, level of lipid peroxidation (LPO). METHODS: One hundred and twenty rats were randomly divided into control group, silica group, and GcAE-treated group. Silicotic animal models were established by direct tracheal instillation of silica into rat lungs surgically. From the second day of model establishment, rats in GcAE-treated group were orally given GcAE [8 g/(kg x d) corresponding to raw herb]. At 7, 14, 21, 28 and 60 days after establishment of the animal model, eight rats in each group were sacrificed, and samples were collected. The malondialdehyde (MDA) content, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities in plasma were assayed by a spectrophotometer. Types I and III collagen were detected by Sirius red polarization and microscopy, and measuered by Image-Pro Plus Version 4.5 for Windows software. RESULTS: GcAE could reduce the lung/body weight ratio of rats exposed to silica, the synthesis of types I and III collagen of the lungs and the level of lipid peroxidation, increase the activities of SOD and GPx. CONCLUSION: GcAE can ameliorate the silica-induced pulmonary fibrosis by increasing the activities of antioxidase and alleviating the damage of lipid peroxidation to the lungs.

ObjectiveTo investigate the role of ATP sensitive potassium (KATP ) channel in protective effects of post-conditioning with hydrogen sulfide against hepatic ischemia-reperfusion (I/R) injury in rats.Methods Thirty adult male SD rats weighing 220-250 g were randomly divided into 5 groups ( n =6 each):sham operation gorup (group S) ; hepatic I/R group; NaHS group; glibenclamide + NaHS group (group G) and 5-hydroxydecanoate (5-HD) + NaHS group (group 5-HD).Hepatic ischemia was produced by 60 min occlusion of left and middle lobe blood flow (about 70％ of the whole liver blood flow) followed by 6 h reperfusion in groups I/R,NaHS,G and 5-HD.NaH S 28 μmol/kg was administered intraperitoneally (IP) at 5 min before reperfusion in groups NaHS,G and 5-HD.Glibenclamide 6 mg/kg and 5-HD 10 mg/kg (KATP channel blockers) were administered IP at 5 min before IP NaHS in groups G and 5-HD respectively.Blood samples were collected at the end of 6 h of reperfusion for measurement of ALT and AST activities and liver specimens were obtained for microscopic examination and measurement of TNF-α content and MPO activity.ResultsLiver I/R significantly increased ALT,AST and MPO activities and TNF-α concentration in group I/R as compared with group S.NaHS administered at 5 min before reperfusion significantly attenuated hepatic I/R injury.Glibenclamide and 5-HD administered before NaHS could block the protective effect of NaHS against hepatic I/R injury.ConclusionHydrogen sulfide postconditioning can protect the liver from I/R injury by activating KATP channel.