Objective:AIM To obtein a recombinant replication-deficient adenovirus expressing LacZ protein. Methods:The recombinant adenovirus vector engineered to express LacZ gene(Ad-LacZ) was constructed by recombination in E.coli-BJ5183.The replication-deficient adenovirus was then packed in HEK293 cells after transfection with the construct and subsequently identified by X-gal staining,electron microscopic observation and dot blotting.The expression of LacZ in HeLa cells infected by Ad-LacZ could be detected by X-gal stain. Results:Recombinant adenovirus expression vector of LacZ was successfully reconstructed.And the titer of Ad-LacZ virus reached 1.58?10~(9) pfu/ml.Under transmission(electron) microscope,adenoviral inclusions could be found in the nucleus of the infected HEK293 cells. Ad-LacZ virus was comfirmed to be replication-deficient by dot blotting.And the expression of LacZ was observed in the infected HeLa cells.The gene transfer capability of the recombinant virus was correlated with multiplicity of infection and the infection time. Conclusion:Recombinant adenovirus expression vector of LacZ was successfully reconstructed.

Objective To study intervention effect of diet habits according to Trans-theoretic Model (TTM) in young and middle-aged patients with asymptomatic hyperuricemia. Methods 120 patients were divided into the intervention group (n=60) and the control group (n=60). The nursing intervention by stages of TTM was implemented to a total of 56 patients in the intervention group in the study. Routine nursing care was taken to 57 patients in the control group. Patients were evaluated by self-designed questionnaire after 1, 3 and 6 months of the intervention in the two groups respectively, including the diets of patients, the change of patients′ eating habit, differences of uric acid, total cholesterol, triglycerides, blood sugar. Results The patients′ intake of beer, seafood, meat, edible oil in the intervention group was lower than the patients′ intake in the control group, while the patients′ intake of fruits and vegetables was higher than the patients′intake in the control group after 1, 3 and 6 months of the intervention, which was statistically significant ( P < 0.01). Compared with 27 cases in the control group, 53 cases in the intervention group were in action phase and maintenance phase after 6-month intervention, the difference was statistically significant (P < 0.01). After six months′intervention, the patients′uric acid, triglycerides in the intervention group was (418.01 ± 24.15)μmol/L and (1.52 ± 0.56) mmol/ L respectively, which was significantly lower than (435.02 ± 26.35) μmol/ L and (2.45 ± 0.74) mmol/L in the control group, t values were-3.47 and 2.20 respectively, P<0.01. Conclusions Diet intervention based on the TTM can effectively promote the change of patients′ unhealthy eating habits, improve self-management skills, and reduce uric acid, which is worthy of generalizing.

Objective To identify the expression of miRNAs in human bone marrow-derived mesenchymal stem cells (hMSCs). Method Low molecular weight RNA fraction (≤200nt) from human bone marrow-derived mesenchymal stem cells was extracted, and polyadenylated by poly(A) polymerase. Then a 5′RNA adapter was ligated to poly(A)-tailed RNA using T4 RNA ligase. RNAs were reversely transcribed and amplified by RT-PCR. The PCR products with a size of about 109 bp were recovered and cloned into pCR 4-TOPO vector. After sequencing, database searching, and expression profiling, the expression of miRNAs in hMSCs was sieved. The expression of novel and some known miRNAs was examined by Northern blotting with small RNAs (≤200nt) isolated from hMSCs, osteosarcoma cell line SOSP-9607 and UMR-106. Results hMSCs were isolated from bone marrow and purified by centrifuge and in vitro culture. Cell markers CD44 were positive, but CD34 and CD45 were negative. A total of 194 clones were subsequently characterized by DNA sequencing and database searching. 52 clones (correspond to 27species) out of 194 clones from hMSCs were identified as miRNAs. One novel miRNAs (PREDICTED-miR-202) was discovered among other 26 known miRNAs, which were predicted in Nature. Northern blotting confirmed that 1 novel miRNAs and 3 known miRNAs (miR-495, miR-34a, miR-17-5p, PREDICTED-miR-202) were stably expressed in hMSCs. Conclusion These findings indicate that a large diverse population of miRNAs are likely important regulators for hMSCs in maintaining their state of self-renewal, and have potential value in stem cells research.

Background and purpose: Primary renal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue(MALT lymphoma) are rare tumors in the kidney.Till now,less than 50 cases have been reported all over the world.This article introduced 2 cases of MALT lymphoma based on the morphological and immunophenotype features,aimed to highlight this rare tumor in terms of clinical characteristics and pathology.Methods:2 cases of renal MALT lymphomas were treated in our hospital.Clinical history were collected,morphological analysis was evaluated based on the HE section,and immunohistochemistry were performed by CD20,CD79a,CD5,CD10,CD43,CD23,BCL10 and cyclinD1 antibodies.Results:2 female patients with the age of 48 and 55,respectively,both of them had a history of chronic pyelonephritis.Under the B ultrasonic and CT scanning,a lump in the kidney was found and whole nephrectomy was performed.In the macroscopic view,tumors were in the renal medulla with dark red color and ill-defined boundary.In the microscopic view,lymphoid cells infiltrate from the pelvis to the medulla of kidney,lymphoid cells mainly consisted of small lymphocytes,centrocyte-like cells,lymphoplasmacytoid and plasma cells.Reactive follicles and lymphoepithelial lesions also could be seen in the tumor.Immunohistochemistry showed that the tumor cells were CD20 and CD79a positive,CD43 was weak positive,but CD5,CD10,CD23,BCL10 and cyclinD1 were all negative.Conclusions:Primary renal MALT lymphomas are very rare diseases.According to the clinical manifestation,it is hard to differentiate from renal cell carcinoma.But the morphological features were consistent with the classic MALT lymphomas in other sites.Immunophenotype profiles were helpful for diagnosis.

Objectives:To study the effects of gene therapy with tissue type plasminogen activator(t PA)cDNA on the formation of thrombo embolism in vascular anastomotic sites. Methods:①The cDNA encoding t PA was amplified by RT PCR using the isolated total RNA as the template from the Bowes melanoma cells.②Recombinant plasmid pAdCMV t PA was cotransfected into 293 cells with pJMa 17 ,and the infectious but replication deficient AdCMV t PA was generated.③The rats were randomly divided into the control and treatment groups.11 0 nylone medical suture was applied to perform rat carotid artery end to end anastomoses.In the treatment group,AdCMV t PA solution was injected into the vascular anastomotic site while AdCMV (no containing t PA DNA) solution was injected into the control group. By means of RT PCR and chromogenic plasmin substrates,the following results were obtained. Results:①The t PA cDNA was successfully cloned and its eukaryotic expressing vector was constructed.②When the isolated RNA was performed with RT PCR,1.69 kb band appeared in the treatment group while the band could not be found in the control group.The t PA activity could be detected postoperatively on the 1st,2 nd,3 rd,4 th,5 th,6 th,7 th,10 th and 13 th day of the treatment,but could not be detected in the control group. Conclusions:The t PA gene can produce t PA having biological activity at anastomotic sites, possibly prevent the formation of thrombus embolism effectively and develop the anastomotic patency.

AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.

Objective To investigated the biological procedure of allograft decalcified bone matrix(DBM)and bone cement(BC)combined with bovine bone morphogenetic protein (bBMP)used for the repair of femoral defect caused by microwave－ induced hyperthermia in dogsby 99mTc－ MDP bone scintigraphy.Method The canine femoral defect(length 25mm,width 10mm)was caused by microwave－ induced hyperthermia(50℃ ,20minutes)and the composite material was implanted .Then the canine femurs were examined by 99mTc－ MDP bone scintigraphy respectively at different postoperative time and the results were compared with that of X－ ray photography and histological observation.Bone cement was implanted in the other femur as a contrast.Results It could be observed at the first and the second month that the radioisotope was gathered in the place where the composite material was implanted and the amount of radioisotope gathered in was the most abundant at the third month and it was lasted to the fourth month. That of the sixth month was decreased to that of the second month.The radiation count of the first, the second, the third the fourth and the sixth month were 93.9± 12.7, 110.7± 16.4,222.1± 24.0,201.3± 26.9 and 111.6± 20.7 respectively,and the count of the third month and the fourth month were more than that of the first, the second and the sixth month(P<0.01).Conclusion The composite material could be remodeled easily and the new bone could be formed by the induction of bBMP. So it could be merged with the normal bone.While the 99mTc－ MDP bone scintigraphy is the object and reliable index to determine the biological procedure of the composite material in dogs.

Objective To investigate the early diagnosis value of arterial blood gas analysis , neuron specific enolase and serum glial fibrillary acidic protein in brain injury of premature infant. Methods In the study, 95 premature infants admitted in our hospital were enrolled. 45 premature infants with brain injury were selected as experimental group. 50 premature infants without brain injury were selected as control group. All patients received arterial blood gas analysis , NSE and GFAP on the 1st day and 7th day after birth. Results There is statistically significant difference between the experimental group and the control group on the 1st day after birth in terms of pCO2, pH, BE and lactic acid (P<0.05). In experimental group, there is statistically significant difference between the 1st day and 7th day after birth in terms of pCO2, pH, BE and lactic acid (P < 0.05). The NSE and GFAP levels had statistically significant difference between the experimental group and the control group on the 1st day and 7th day after birth(P < 0.01); the NSE levels of experimental group had statistically significant difference between the 1st day and 7th day after birth (P < 0.05); The GFAP levels of experimental group had statistically significant difference between the 1st day and 7th day after birth (P < 0.01). Conclusion pCO2, NSE and GFAP levels are correlated with brain damage in premature infants in early stage. This could provide evidence of early diagnosis for brain injury in preterm infants.

Objective To clarify the temperature curve of the irradiation target area,its adjacent tissue and the whole body during extracorpereal microwave irradiation, then to compare and optimize different irradiation models. Methods Different parts of the chest of adult New Zealand white rabbit were irradiated using different extracorporeal microwave irradiation models. The temperature of the irradiated skin, the subcutaneous and deep parts, the adjacent tissues and the anus was measured. The experiment was bi-factor and multi-level designed according to the repeatedly measured data and the rabbits was divided into group a,b,c and d. Results The increase rate of the surface temperature in the dorsal lung was similar between group d and group b1(F=10.04,P<0.01). However,the increase rate of the surface temperature in the ventral lung of group d was lower, and the mean temperature of this site measured 10 minutes later was also lower than group b1(F=10.04,P<0.01). The increase rate of the rectal temperature of group d was higher,and the mean rectal temperature tested 10 minutes later was also higher than group b1(F=7.04,P<0.01). Conclusions Multi-array irradiation could achieve satisfactory irradiation depth and appropriate therapeutic temperature. Well controlled extracorporeal microwave irradiation under is an ideal thermotherapy method.

Objective To investigate the role of mylin basic protein,S100B and arterial blood gas analysis's levels in early diagnosis of brain injury in premature infant.Methods A total of 95 premature infants treated in our hospital were enrolled in the study.Experimental group was 45 premature infants with brain injury.Control group was 50 premature infants without brain injury were the.All patients were detected with arterial blood gas analysis,MBP and S100B on the 1st day and 7th day after birth.Results The pH,PCO2,BE,lactic acid,MBP and S100B's levels in experimental group were significantly different between the 1st day and 7th day after birth.In the 1st day after birth,compared with the control group,the pH,PCO2,BE,lactic acid,MBP and S100B in the experimental group were obviously high than that of control group.Conclusion On the 1st day after birth,monitoring arterial blood gas analysis,S100B protein and MBP's levels could be useful in early diagnosis of brain injury in preterm infants.