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.

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.

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.

Objective: To examine whether there are differences between dependent and independent individual’s shame and sad memories. Methods: Self -report measures were used. Results: ① There were significant interactions between emotion and cognitive style of the number of words recalled, number of times that others were mentioned. ②Field dependent individuals mention more emotional behaviors and presence of others. ③The subjects mentioned more emotion words and time in sad memories. ④In shame memories, there were less self-references than others-references, marginally less self in thinking than self in other descriptions. ⑤In sad memories, field dependent individuals reported less selfreferences than others-references, and less self in thinking than self in other descriptions. ⑥There were more emotional behaviors and self-thinking in sad memories than in shame memories. Conclusion: Field dependent and independent individuals demonstrate differences in emotional memory, and the difference is emotion specificity.

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.

BACKGROUND:The treatment of infected nonunion is a difficult problem in orthopedic trauma, which needs to completely eliminate foci of osteomyelitis to control infection, repair soft tissue defect and close the wound, repairs bone defects to promote healing and protect limb function as much as possible. Autogenous micromorselized bone can be easily obtained and recombinant human bone morphogenetic protein-2 has variety of sources. Their mixed grafting mode is safe and reliable in repairing of bone defects, which are suitable for repair of infected nonunion. ＜br> OBJECTIVE:To study the operating method and clinical results of repairing infected nonunion using autogenous micromorselized bone grafting mixed with bone morphogenetic protein-2. ＜br> METHODS:Twenty-three cases underwent infected nonunion repair with autogenous micromorselized bone (l ium, long bone metaphyseal cancellous bone) mixed with bone morphogenetic protein-2, including 7 cases of femoral bone defect caused by osteomyelitis after debridement and external fixation and 16 cases of tibial bone infected nonunion. Eleven of 23 cases continued to use the external fixation because of scar contraction or poor conditions of soft tissue, and the other 12 cases switched to use internal fixation. X-tray was taken to evaluate the bone healing at the 1st, 3rd, 6th and 12th months after operation. The function of the affected extremity was assessed by Johner-Wruhs grade. ＜br> RESULTS AND CONCLUSION:Al of the cases were averagely fol owed up for 13.4 months. The bone defects and bone fracture healed in al cases without infection, and the wounds were al sealed. The excellent and good rate according to Johner-Wruhs grade was 95%. These findings indicate that the bone grafting mode of autogenous micromorselized bone mixed with bone morphogenetic protein-2 is characterized as variety of sources, high safety and reliable osteogenic ability in repair of bone defects.

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)