The serum soluble interleukin 2 receptor(sIL—2R)in 39 patients with malignant tumour of degestive system wasemeasured using by sandwith ELISA with monoclonal and polycional antibodies technigue.The results show the level of sIL—2R was obvious higher than level of sIL—2R of the controls(P

Moss has high tolerance and accumulating capacity to heavy metal.In this study, the distribution of heavy metal elements in moss sampled from lead-zinc mine was analyzed by X-ray fluorescence spectrometry.The speciation of lead was analyzed by X-ray absorption near-edge spectroscopy.Research showed that the contents of Pb, Zn, Cd and As in the moss of the mining area were extremely high, and their maximum concentration were 1.06 mg/g , 1.23 mg/g, 30.5 μg/g, 13.2 μg/g, respectively.Besides, the shoots especially the new tissue of the moss were the major sites for accumulation and storage of heavy metals.The micro-distribution characteristics varied among Hypnum plumaeforme and Brachytheciumprocumbens, indicating the difference of different species of moss in absorption pathway, accumulation and tolerance mechanisms for heavy metal.Linear combination fitting results indicated that the main lead speciation in moss was lead phosphate (78%) and lead oxide (22%), which suggested that the precipitation of lead phosphate might be the main mechanism of tolerance for moss.

BACKGROUND: The particular bystander effect of suicide gene can remarkably kill tumor cells. Meanwhile, it can be used together with radiotherapy as well as immune gene therapy, and overcome the defect of low gene transduction efficiency.Cytosine deaminase (CD) can generate a powerful bystander effect.OBJECTIVE: To observe the effect of a eukaryotic expression plasmid plRES2-AcGFP1-CD mediated by liposome transfected into bone marrow mesenchymal stem cells (BMSCs) and its gene expression.DESIGN, TIME AND SETTING: A cytologic experiment at genetic level was performed at Research and Development Center of Stem Cell and Tissue Engineering, Dalian University of Technology from May to December 2007.MATERIALS: A total of 6 C57BL mice of SPF degree and weighing 18-20 g were used for separation and culture of BMSCs.Escherichia coli DH5a was provided by Research and Development Center of Stem Cell and Tissue Engineering, Dalian University of Technology. Lipofectamine~(TM) 2000 was provided by Invitrogen, China.METHODS: The DNA plasmids were extracted from transformed Escherichia coli DH5a. plRES2-AcGFP1-CD plasmid was identified by BamHI/Xhol double digestion. The BMSCs derived from mouse femur and tibia were cultured and purified by adhesion method in vitro. Signal cell suspension prepared by BMSCs of the third generation was cultured by adding fluorescence-labeled CD44, CD45, CD90 and CD105 antibodies. BMSCs of the third generation were transfected by LipofectamineTM 2000 mediation.MAIN OUTCOME MEASURES: Identification of recombinant plasmids; the expressions of surface markers on BMSCs were detected by flow cytometry; the expressions of cytosine deaminase gene were observed at 36 and 48 hours after transfection under fluorescent inverted microscope.RESULTS: After agarose gel electrophoresis, a band appeared at 1.0-1.5 kb of the digested products of plRES2-AcGFP1-CD plasmids, and the band was accorded with the length of cytosine deaminase gene in the length. Flow cytometry demonstrated that the cells were negative for CD45 but positive for CD44, CD90 and CD105. Fluorescent inverted phase contrast microscope suggested that at 36 hours after plRES2-AcGFP1-CD gene transfection an expression of green fluorescent protein was found in the BMSCs; additionally, at 48 hours after transfection, the expression of green fluorescent protein remained and the intensity was remarkably increased.CONCLUSION: The liposome-mediated plRES2-AcGFP1-CD gene successfully expressed in BMSCs, and the expression peaked at 48 hours after transfection.

AIM:Activin A,a member of transforming growth factor superfamily,is the negative regulator factor in liver regeneration. In this study,the effects of extract of Ginkgo Biloba on hepatic fibrosis and the expression of Activin A in rats with cirrhosis were investigated. METHODS:The experiment was performed at in the Central Laboratory of Wuhan First Hospital from September 2005 to December 2006. ①Thirty-six male SD rats of(160?20) g were randomized into 3 groups:control group,model group and treatment group. ② Except the rats in the control group,others were intraperitoneally injected with 500 mL/L CCl4 for 8 weeks to establish models of hepatic fibrosis. Meanwhile,the extract treatment group was infused with the extract of Ginkgo Biloba(Chinese drugs preparation laboratory of Wuhan First Hospital,detected by Hubeu Wushi Medicine Industry Co.,Ltd. No. 02-391) daily for 8 weeks. ③After administration,all anesthetized rats were sacrificed. Blood samples were collected for the determination of liver function biochemical indexes. Liver tissue samples were used for histopathological examinations. The expression of Activin A was determined by immunohistochemistry and RT-PCR. RESULTS:All 36 rats were involved in the final analysis. ①The liver function in extract treatment group was significantly improved compared with that in model group. ②The grade of fibrosis in extract treatment group were remarkably lower than that in model group under light microscope. ③The positive staining of Activin A in treatment group was significantly reduced compared with model group. ④The expression of Activin A mRNA in extract treatment group was significantly reduced compared with model group. CONCLUSION:Extract of Ginkgo Biloba can effectively decrease the expression of Activin A in rats with hepatic fibrosis caused by CCl4,and lessen the degree of hepatic fibrosis.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are easily isolated and amplified, and facilitate the exogenous gene transfer and expression. In the human medicine, it is believed that BMSCs are ideal therapeutic cells and target cells in the gene therapy.OBJECTIVE: To investigate liposome-mediated cytosine deaminase (CD) gene transfecting rabbit BMSCs and its gene expression. DESIGN: A single sample observation. SETTING: Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University.MATERIALS: This study was performed at in the Dalian Research and Development Center for Stem Cell and Tissue Engineering; Department of Biochemistry, College of Basic Medical Science, Dalian Medical University from March 2006 to June 2007. New Zealand big-ear white rabbits of either gender, weighing 2.0-2.5 kg, with the age of 5 months old, were included in this study. METHODS: The CD gene was obtained from E.coli JM109 DNA by polymerase chain reaction (PCR). The fragment was cloned into pMD19-T vector. After restriction enzyme BamHI/XhoI digestion analysis and DNA sequence analysis, pIRES2-AcGFP1-CD eukaryotic expression plasmid was constructed. Meanwhile, BMSCs were harvested, cultured and identified. After enzyme digestion of eukaryotic expression plasmid, the rabbit BMSCs were transfected by Lipofectamine 2000-mediated method. Twenty-four hours after transfection, expression of green fluorescent protein was observed under an inverted fluorescent microscope. MAIN OUTCOME MEASURES: Construction of eukaryotic expression plasmid and identification of CD gene-transferred BMSCs. RESULTS: CD gene was cloned and connected to eukaryotic expression plasmid with green fluorescence. Twenty-four hours after transfecting rabbit BMSCs, it was found under an inverted microscope that under the excitation of 488 nm blue light, green fluorescence appeared in the pIRES2-AcGFP1-CD and pIRES2-AcGFP1 empty-plasmid transfected BMSCs, but not in the non-transfected ones. It indicates that CD gene successfully transferred BMSCs. CONCLUSION: BMSCs are ideal vectors in the CD gene therapy.

ObjectiveTo investigate the mechanism of anti-inflammatory effect of diallyl sulfide (DAS) in protection against acute lung injury (ALI) in rats with paraquat poisoning.Methods Eighty male Wistar rats were randomly divided into four groups, namely: control group, model group, dexamethasone (DXM) treatment group, and DAS treatment group, with 20 rats in each group. The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by gavage, while the same volume of normal saline (NS) was given in same manner in control group. 100 mg/kg of DAS, the same volume of NS, or 1 mg/kg DXM injection were given respectively in DAS treatment group, model group, or DXM treatment group intraperitoneally after exposure to paraquat, once a day for 14 days. Five rats in each group were sacrificed at 1, 3, 7, 14 days, respectively. The inferior lobe of right lung was harvested, and the degree of lung injury was observed with hematoxylin and eosin (HE) staining under optical microscope; the upper lobe of right lung was used to determine the lung wet/dry weight (W/D) ratio and for evaluation of the degree of pulmonary edema. The expression of nuclear factor -κB (NF-κB) in the middle lobe of right lung was assessed with immunohistochemistry. The expression of tumor necrosis factor -α (TNF-α) mRNA in the left lung was determined with the reverse transcription-polymerase chain reaction (RT-PCR).Results① The pulmonary structure in control group was found to be intact. However, in the model group there were progressive pathological changes in lung, including marked edema and thickening of alveolar walls, collapse of alveoli, infiltration of inflammatory cells, alveolar wall, and obvious bleeding in the local lung tissue, and formation of transparent membrane in alveolar space. Less infiltration of inflammatory cells and no obvious destruction were found in alveolar structure in the DAS and DXM treatment groups.② Lung W/D ratio: lung W/D ratio of model group was apparently higher than that in control group at every time point, and peaking on the 3rd day (6.15±0.54 vs. 4.15±2.10,P< 0.05), and the ratio of lung W/D of DAS and DXM treatment groups was obviously lower than that in model group at every time point, especially on the 3rd day (3.99±1.26, 4.30±0.70 vs. 6.15±0.54, bothP< 0.05), but there was no significant difference between DAS and DXM treatment groups in this regard.③ The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell nuclei of the control group, while extensive NF-κBp65 expression was found in model group. Minimal NF-κBp65 positive expression in the cytoplasm and even less positive expression in the nucleus was found in the DAS and DXM treatment groups, and integralA value was significantly lower in the DAS and DXM treatment groups than that of the model group, especially on the 3rd day [(17.98±0.06)×107, (18.53±0.04)×107 vs. (28.85±0.61)×107, bothP< 0.01], but there was no significant difference between DAS and DXM treatment groups.④ It was shown by RT-PCR that the expression of TNF-α mRNA in lung tissue of the model group was significantly higher than that in the control group on the 3rd day (gray value: 3.63±0.62 vs. 0.51±0.13, P< 0.05). The expression of TNF-α mRNA in lung tissue was significantly decreased in DAS and DXM treatment groups compared with model group (gray value: 2.49±0.57, 2.02±0.26 vs. 3.63±0.62, bothP< 0.05), but there was no significant difference between DAS and DXM treated groups.ConclusionTreatment with an intraperitoneally injection of DAS is capable of attenuate the extent of PQ-induced ALI in rats by alleviating pulmonary edema, inhibiting the expression of NF-κB and TNF-α in lung tissue, and ameliorating pathological changes in lung tissue.

Objective To prepare polyclactic-co-glycolic acid(PLGA) polymer ultrasound contrast agent loading sudan black dye (SB-PLGA) and study the enhancement effect of it on rabbit lymph nodes imaging and as a sentinel lymph node (SLN) biopsy in the feasibility and value of the indicator.Methods SB-PLGA ultrasound contrast agent was made by double emulsion and freeze-drying methods.Thigh subcutaneous rabbit VX2 tumor model of eight lymph nodes were set up with the foot pad subcutaneous injection of contrast agents to get contrast-enhanced ultrasound imaging of the popliteal lymph node and the second inguinal lymph node stations.Then the blue stained lymph nodes were resected by popliteal lymph node dissection and the second station lymph nodes were also observed after 30 mins respectively,lymph nodes were examined by frozen sections HE staining in parallel.Results SB-PLGA microbubbles had a tight size distribution.Ultrasound imaging can be significantly enhanced lymph node imaging,the second station lymph node imaging was not obvious.Popliteal lymph node dissection for lymph node staining,30minutes later,frozen sections HE staining were seen within the lymph node contrast agent present in the lymphatic sinus and a large entry of macrophages.Inguinal lymph node dissection showed the second station of lymph nodes no blue staining.Conclusions SB-PLGA ultrasound contrast agent is good for sentinel lymph node imaging and biopsy indicator.Contrast agent retention and accumulation in the lymph nodes by the uptake of macrophages may be associated with the mechanisms of lymph node enhanced imaging.

Objective To prepare a kind of superparamagnetic iron oxide (SPIO) and doxorubicin loaded multifunctional polymer microbubbles (MPMBs),and to explore its potential application as an ultrasound(US)/magnetic resonance (MR) imaging contrast agent in vitro.Methods The MPMBs and normal polymer microbubbles (PMBs) were made by double emulsion and freeze-drying methods.The physical property,drug encapsulation efficiency and the drug-loading efficiency of MPMBs were determined,and the release property and US/MR imaging enhancement of MPMBs were observed.Results The MPMBs had a regular shape and narrow size distribution.The drug encapsulation efficiency was (60.20±2.69) ％,and the drug-loading efficiency was (6.02 ± 0.27) ％.The in vitro release experiment showed that ultrasound can promote the release of doxorubicin in MPMBs.US imaging in vitro showed that the enhancement of MPMBs was better than PMBs,and MR imaging in vitro conformed that MPMBs could well enhance MR imaging.Conclusions The MPMBs is a multifunctional contrast agent with the treatment function as well as US/MR dual-mode imaging enhancement effect.

[ABSTRACT]OBJECTIVETo study of the relationship of age and climate with the nasal mucosa lesions.METHODSAll the patients met with the inclusive criteria were registered and followed-up once a month and their nasal mucosal lesions were observed through nasal speculum and/or nasal endoscope. The recording data was analyzed by SPSS17.0 software.RESULTSA total of 259 patients, 555 person-times and 957 lesion spots were observed. The nasal mucosa was injured by airflow with the highest incidence rate from October to next year April. Patients numbers gradually increased after August, and significantly increased at November, December (P<0.05). Children aged from 2 to 12 years old and the adults aged from 40 to 80 years old were susceptible population (P<0.05), the incidence rate had a negative relation with humidity (P<0.05). The susceptible spots of the lesion were ranked as follows: Little area, inferior turbinate, mucosa of the both side of the deviated nasal septum, inferior meatus, middle meatus, middle turbinate and olfactory area. CONCLUSIONNasal mucosal injury relates to age and seasons. The nasal cavity airflow may be a factor of these lesions.

T lymphocyte subpopulation and level of soluble interleuckin 2 receptor in peripheral bloodand serum of primary hepatocellular carcinoma and donor were tested.The results show that de-crease of T lymphocyte subpopulations is related with increase of level of soluble interleuckin2 re-ceptor in patients with primary hepatocellular carcinoma.