Objective Tiam1, a member of guanine nucleotide exchange factors, plays an important role in tumor invasion and metastasis.This study aimed to construct Tiam1 truncated recombinant plasmids and induce the expression of GST-tagged human Tiam1 fusion proteins in Escherichia coli (E.coli), followed by purification and identification of the GST-Tiam1 fusion protein.Methods The cDNA fragments of Tiam1 C685, C751 and C1199 were amplified by PCR and cloned into the pGEX-4T-1 vector.After verified by DNA sequencing, the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells, and the expression of the fusion protein was induced by IPTG.The GST-tagged human Tiam1 fusion proteins were purified with glutathione-agarose resin and indentified by Western blot.Results Three Tiam1 truncated recombinant plasmids were constructed successfully.The recombinant fusion proteins GST-Tiam1 C685, GST-Tiam1 C751, and GST-Tiam1 C1199 were expressed mainly in the form of soluble proteins in the cell lysate supernatant with expected relative molecular weight of 100, 108, and 157 kD.Conclusion The recombinant plasmids expressing the bioactive fusion proteins were constructed successfully, which has prepared the ground for the subsequent studies of the Tiam1 protein.

Objective To investigate the imaging features of pancreatic acinar cell carcinoma (PACC),and to assess the role of CT and MRI in the diagnosis of the disease.Methods CT and MRI data of 7 cases with PACC confirmed by surgery and pathology were reviewed retrospectively.Plan and dynamic contrast-enhanced CT were performed in 4 cases.MRI with T1 WI,T2 WI,and dy-namic contrast-enhanced series were performed in 3 cases.Results All of the PACC lesions were manifested as a single solitary mass.1 lesion was located in the pancreatic head,and the other 6 in the pancreatic body-tail.On plan CT,all of the 4 lesions ap-peared hypodense and 3 lesions had irregular more hypodense region in the lesions.On the contrast-enhanced CT,the tumor paren-chyma showed mild to moderate enhancement with non-enhanced hypodense region in the arterial phase,and lower enhancement than that of the surrounding normal pancreatic tissue in the portal and delayed phase.All of the 3 cases were heterogeneous hypointensity on T1 WI and hyperintensity on T2 WI.The manifestations of the tumors on contrast-enhanced MRI were similar with that on the contrast-enhanced CT.Dilation of the pancreatic duct was seen in 4 cases.Liver metastasis was seen in 1 case.Surrounding tissues were invaded in 4 cases.Conclusion CT and MRI can display the features of PACC and help to improve the diagnostic accuracy.

This paper dealt with the influences of different media, temperature, light and pH to the amount of oospore formation. The results showed that millet agar, bai yun beau agar and soybean agar provided the favorable condition for oospore formation, the number of oospores was 131.6~149.6 /cm2, but a few oospores was formation on frozen pea agar. The results also suggested that the optimal conditions for oospore formation were 18℃, pH7 and darkness. The oospores failed to be formed tinder fluorescent or black light.

Objective To identify the imaging performance and differences between type] and type Ⅱ papillary renal cell carcinoma (PRCC).Methods Data of 21 lesions of type Ⅰ,27 lesions of type Ⅱ (1 patient had 2 lesions) in 47 patients was retrospectively analyxed.All patients with pathologically proven PRCC were examined by contrast CT or MRI preoperatively.The morphological features,outside invasion signs and performance on contrast-enhanced CT were compared by qualitative and quantitative studies.The maximum diameter of tumors and CT values,△CT values in corticomedullary and nephrographic phase were analyzed by two-sample t-test,classified variable were compared by the Pearson X2 test or the Fisher exact test.Results On morphological behaviors,type Ⅱ PRCC were significantly larger than type Ⅰ PRCC (t =-2.604,P =0.013),more heterogeneous (X2 =14.928,P =0.000),greater probability to show cystic degeneration or necrosis (X2 =5.598,P =0.018) with more severity (X2 =4.769,P =0.029).There was no significant difference in hemorrhage and calcification between the two types observed by contrast-enhanced CT.Respectively,66.7 ％ of type Ⅱ PRCC and 23.8％ of type Ⅰ PRCC had papillary nodule,with obviously significant difference (X2 =8.694,P =0.003).In outside invasion signs,except for margins,type Ⅱ had more easily invaded peripheral fat,renal sinus and distant metastasis compared with type Ⅰ (P＜0.05).On contrast enhanced CT,there were significant differences in CT values and △CT values in corticomedullary phase between the two types (t =-2.674,P =0.012;t =-3.109,P =0.005).And there were no significant difference in unenhanced and nephrographic phase.Conclusions There were certain difference in morphological features,outside invasion signs and enhancement degree between type Ⅰ and type Ⅱ PRCC,and part of type Ⅱ PRCC had aggressive biological behaviors with worse prognosis.

Objectives:To study the relationship between nutrition status and renal function in renal disease patients. Methods: Body weight, height and the outcomes of blood biochemistry and routine were analyzed in 110 renal patients. Results: Body weight distribution in renal failure group had marked difference compared with nomal renal function group, and the number of patients whose actual body weight being lower than ideal body weight was obviously increased. The levels of RBC, Hb and TLC were significantly decreased(P

Chronic activation of the B-cell antigen receptor (BCR) signaling pathway performs a critical function in the pathogenesis of numerous subtypes of B-cell malignancies and transforms normal cells into malignant cells. Bruton tyrosine kinase (BTK) is a member of the TEC family of tyrosine kinases and is a key regulator of the BCR signaling pathway. BTK inhibition has emerged as a new target for therapeutic intervention in B-cell malignancies. This review summarizes recent developments of BTK inhibitors in B-cell malignan-cies.

Objective To investigate the distribution of urate crystal as well as the relationship bet ween the features of the crystals and the attacks of joint pain and/or swollen by foot dual-energy CT.Methods Eight-four patients (68 were diagnosed as gout, 11 were patients with hyperuricemia and 5 were diagnosed as other types of arthritis) who recently experienced foot swelling and/or pain were enrolled and all of them were performed foot dual-energy CT.The relationship between the features of the urate crystals and the attacks of gouty arthritis was determined by Chi test and the potential risk factors were identified by Logistic multiple regression analysis.Results Two hundred and seventyeight urate crystal depositions were found in 68 gout patients,and the most common deposition sites were the distal parts of the first toe(18.2％),the first metatarso-phalangeal joint ( 16.8％ ),calcaneus ( 17.5％ ),the lower end of tibia ( 11.8％ ).Furthermore,patients with the urate crystals deposited in the first metatrasophalangeal joint or the lower end of of tibia were more likely to experience acute episodes of gout attack (P＜0.01,P＜0.05 respectively).In addition,the shape,size and quantity of urate crystals also affected episodes of acute attack of gout.Conclusion Dual-energy CT,which is a non-invasive method,could clearly reveal urate crystal depositions and is helpful for the diagnosis and follow-up of patients with gout.The location,shape,size and quantity of urate crystals and soft tissue swelling,bone erosion may affect the acute attack of gout.

OBJECTIVEThis paper aimed to determine the mRNA expression of osteoclast-related factors interleukin-6 (IL-6), interleukin-12 (IL-12) p35, IL-12p40, matrix metalloproteinase-9 (MMP-9), nuclear factor of activated T-cells cytoplasmic 1 (NFATcl), receptor activator of nuclear factor-KB (RANK), and tumor necrosis factor-α (TNF-α) mRNA in murine macrophages infected by a periodontitis patient's own tissue nucleic acid. Another aim was to investigate the effects of a periodontitis patient's own tissue nucleic acid on the differentiation of macrophages into osteoclasts.

METHODSInflammatory periodontal tissue samples of chronic periodontitis patients were taken during periodontal flap surgery, and healthy gingival tissue samples were taken from orthodontic patients during tooth extractions. Total RNA from periodontal tissue was extracted and reversely transcribed into cDNA and then cryo-preserved until further use. First, specific sequence oligodeoxynucleotide MT0I at a concentration of 1 µg · mL⁻¹ was added in murine macrophage RAW264.7, and the cells were incubated for 3 hours. Cells with PBS (1 µg · mL⁻¹) were used as negative controls. The inflammatory periodontal tissue cDNA and healthy periodontal tissue cDNA (1 µg · mL⁻¹) was added subsequently. There were four experimental groups: healthy periodontal tissue cDNA+ RAW264.7, inflammatory periodontal tissue cDNA+RAW264.7, MT01+healthy periodontal tissue cDNA+RAW264.7, and MT01+inflammatory periodontal tissue cDNA+RAW264.7. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of osteoclast-related factors IL-6, IL-12p35, IL-12p4O, MMP-9, NFATcl, RANK, and TNF-α mRNA after 3, 6, 12, and 24-hours.

RESULTSThe mRNA levels of osteoclast-related factors NFATc1, MMP-9, TNF-a, IL-6, IL-12p40, IL-12p35, and RANK in RAW264.7 were markedly upregulated with the treatment of periodontitis patient's own tissue nucleic acid. However, the mRNA expression of osteoclast-related factors was inhibited by use of an immunosuppressant MT01.

CONCLUSIONThe periodontitis patient's own tissue nucleic acid could promote the differentiation of murine macrophage into osteoclasts.

Animals ; Cell Differentiation ; Cytokines ; metabolism ; Gene Expression ; Gingiva ; Humans ; Interleukin-12 Subunit p40 ; Interleukin-6 ; Macrophages ; Matrix Metalloproteinase 9 ; Mice ; Osteoclasts ; metabolism ; Periodontitis ; RNA, Messenger ; Tumor Necrosis Factor-alpha

OBJECTIVE:To study the antitumor activity of Anthopleura xanthogrammica crude extract on human lung cancer SPC-A1 cells in vitro. METHODS:A. xanthogrammica crude extract obtained by the methods of repeated freezing and thawing,ac-etone precipitation. After treated with crude extract 0(blank control),0.625,1.25 and 2.5 mg/ml for 24,48 and 72 h,the activity of SPC-A1 cells were measured by MTT assay. The growth inhibition rate and IC50 were also calculated. 24 h later,the morphologi-cal changes of SPC-A1 cells were observed by HE staining and AO/EB fluorescence staining. RESULTS:MTT assay showed that A. xanthogrammica crude extract has significant inhibitory effect on the proliferation of human lung cancer SPC-A1 cells;with the increasing of the concentration and the extension of the time,the inhibitory rate was increased. Its 24 h,48 h ,72 h IC50 were 1.81,1.32 and 1.18 mg/ml. HE staining and AO/EB staining appeared obvious morphological changes of apoptosis that cell mor-phology narrowed,vacuoles arose in the cytoplasm,karyopyknosis and part of nuclear disappearance occurred. CONCLUSIONS:A. xanthogrammica crude extract has an inhibitory effect on the proliferation of human lung cancer SPC-A1 cells.

Objective Study the concentration of the hepatocyte growth factor(HGF) the expression product of recombinant human hepatocyte growth factor naked plasmid ,and whether the body generates immune responseafter wsing HCF. Methods Selected 21 patients with severe ischemic disease of lower extremity (Rutherford classification 4-6 class) , after signing the informed consent and divided them into four dosage groups( 4 mg, 8 mg, 12 mg and 16 mg) random.In each group the dosage was the lower limbs partly intramuscular injection following the feeding artery in experimental stage Dl and D15. The plasma samples were collected to determine the HGF concentration in Dl ( before administration), D8, D15 ( before administration) , D21, D59, and the HGF-antibody concentration in Dl, D15, D28, D59, D91. Results (1)The concentrations of HGF in the subjects ranged from 216 pg/mL to 1189.75 pg/mL.(2) HGF-antibodies were not dectected in the subjects' plasma. Conclusions After using recombinant human HGF naked plasmid through intramuscular injection, the concentration of HGF expression product in subjects' peripheral blood does not have abnormally changed and using the drug the human body immune response does not generate.