Objective To express and purify the killer immunoglobulin-like receptor KIR3DL1 extracelluar domain.Methods pUC57-KIR3DL1 was used as template,KIR3DL1 extracelluar domain was amplified by polymerase chain reaction(PCR)and cloned into pGEM-T vector with A-T cloning technique.After DNA sequence analysis,the target fragment was inserted into the prokaryotic expression vector pET28a-DsbA to construct the recombinant vector pET28a-DsbA /KIR3DL1.The recombinant plasmid was transformed into E.coli BL21(DE3),and induced with IPTG.Bacterial pellets were resuspended in 8M urea and centrifuged to remove the insoluble material.The crude extract was purified by passing over a Ni-NTA-agarose column.After the inclusion body flowing through the Ni-NTA-agarose affinity chromatography was refolded successfully,it was purified by Superdex75 gel filtration and the purification effects of the fusion protein were identified by SDS-PAGE and western blot.Result Some fusion proteins were expressed in the supernatant,and the others were expressed in the form of inclusion bodies.The purity of fusion protein was over 95% after purification under denaturing condition.Conclusion The highly efficient expression of KIR3DL1 extracelluar domain laid the foundation for the further studies on exploration of the mechanism of immunization recognition between KIR3DL1 and its ligand.

Objective To investigate the correlation of clinicopathological features with osteopontin and aquaporin-1 expres-sion in laryngeal carcinoma.Methods From August 2011 to December 2014 in Dongfeng General Hospital,80 laryngeal cancer specimens and adjacent normal tissue samples were selected,all samples were carried out osteopontin and aquaporin Immunohistochemical analysis expression and clinicopathological characteristics and correlation analysis.Results Osteopon-tin staining particles and aquaporin-1 were located in the stromal cells,yellow dye.In cancer tissue,osteopontin and aquapor-in-1 positive granules were mainly located in the cytoplasm of cancer cells,diffuse shading,coloring deeper place near nests. The positive expression rates of osteopontin and aquaporin-1 in the laryngeal carcinoma were 70.0% and 56.3% respective-ly,the positive rates in the adjacent tissues were 5.0% and 7.5% respectively that compared to the differences were statisti-cally significant (P <0.05).In 80 cases of laryngeal carcinoma.The expression of osteopontin and aquaporin-1 in different clinical stages,differentiation and lymph node metastasis tissues were statistically significant (P <0.05).Spearman correla-tion analysis showed that the osteopontin and aquaporin-1 in laryngeal carcinoma were positively correlated (P <0.05).Con-clusion Osteopontin and aquaporin-1 expression in laryngeal carcinoma were showed high status,and the clinical stage,de-gree of differentiation and lymph node metastasis were significantly correlated that can work together to promote tumor inva-sion and metastasis.

Background:Colorectal cancer is one of the commonly seen malignant tumors in digestive system. Most patients with colorectal cancer were diagnosed in advanced stage and the prognosis is poor. Early diagnosis is crucial to improve the overall survival of patient. Currently,molecular markers for diagnosis of colorectal cancer has attracted more and more attention of the investigators. Aims:To investigate the expression and significance of Rab5A in colorectal cancer. Methods:Colorectal cancerous tissue and the paired para-cancer noncancerous tissue of 135 patients who underwent resection for colorectal cancer were collected. Expression of Rab5A mRNA in cancerous tissue,para-cancer noncancerous tissue,colon cancer cell lines HCT116,HT-29,LoVo and SW480 and normal colonic epithelial cell line NCM460 was determined by real-time PCR. Protein expression of Rab5A in cancerous tissue and para-cancer noncancerous tissue of 4 patients was detected by Western blotting. Protein expression of Rab5A in cancerous tissue of 110 patients was detected by immunohistochemistry. Correlations between expression of Rab5A and clinicopathological characteristics and prognosis of colorectal cancer were analyzed. Results:Expression of Rab5A mRNA in cancerous tissue was significantly higher than that in para-cancer noncancerous tissue(P=0. 003). Expression of Rab5A mRNA in colon cancer cell lines HCT116, HT-29,LoVo and SW480 was significantly higher than that in normal colonic epithelial cell line NCM460(P﹤0. 05). Protein expression of Rab5A in cancerous tissue was significantly higher than that in para-cancer noncancerous tissue. In 110 cancerous tissues,51(46. 4%)had low expression of Rab5A and 59(53. 6%)had high expression of Rab5A. Protein expression of Rab5A was significantly correlated with tumor size,serum carcinoembryonic antigen(CEA)level and TNM stage(P =0. 008;P =0. 002;P =0. 010). 3-year survival rate of patients with high Rab5A expression was significantly lower than that of patients with low Rab5A expression(52. 1% vs. 77. 5%,P ﹤0. 05). Rab5A protein expression,serum CEA level and TNM stage were correlated with the prognosis of patients with colorectal cancer( P =0.009;P =0. 006;P =0. 017)and were independent factors for prognosis(P =0. 026;P =0. 032;P=0. 014). Conclusions:Expression of Rab5A correlates with the malignant behavior of colorectal cancer and its high expression indicates a poor prognosis. Rab5A might be used as a predictor for prognosis of colorectal cancer.

Objective To assess the cli ni cal usefulness of dual-modality positron emission tomographic (PET)-computed t omographic (CT) imaging in pulmonary lesions. Methods Fo rty-nine patients with pulmonary lesions examined with ?~2 test. Results There were 50 primary lesions in 49 patients. Accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predict ive value (NPV) were 90.0%, 95.2%, 62.5%, 93.0%, and 71.4%, respectively wi th PET-CT, 74.0%, 76.2%, 62.5%, 91.4%, and 33.3%, respectively with CT. There was significant differences between them (P0.05). Conclusion PET-C T has an important clinical value in differentiating benign lesion from malignan cy and staging in lung cancer.

A pot experiment was conducted to study effect of drought stress on leaf physiological characteristics and growth of one year old Stellaria dichotoma seedlings. The result showed that plant height and shoot dry weight significantly decreased with decrease in soil water content; however, root length and root dry weight increased at light drought stress and decreased at severe drought stress. The result also showed that with the decrease of soil water content, proline content in S. dichotoma leaves decreased then increase, while solube protein content decreased. Activities of SOD and POD in S. dichotoma leaves significantly decreased as soil water content decreased, while activity of CAT significantly decreased at severe drought stress. Membrane permeability in S. dichotoma leaves increased, while MDA content decreased then increased as soil water decreased. These results suggest that S. dichotoma had osmotic stress resistance ability and reactive oxygen scavenging capacity at light drought stress, which caused S. dichotoma growth was no inhibited at a certain extent drought stress.
Droughts ; Plant Leaves ; enzymology ; growth & development ; metabolism ; Plant Proteins ; metabolism ; Plant Roots ; enzymology ; growth & development ; metabolism ; Proline ; metabolism ; Seedlings ; enzymology ; growth & development ; metabolism ; Stellaria ; enzymology ; growth & development ; metabolism ; Water ; metabolism

Humans ; Laryngoscopy ; Larynx ; Magnetic Resonance Imaging ; Tomography Scanners, X-Ray Computed ; Tuberculosis, Laryngeal ; diagnosis

Objective To develop an HPLC method for the measurement of n-3 fatty acid index of serum cholesteryl esters.Methods Serum triglycerides were hydrolyzed with ethanolic sodium hydroxide and cholesteryl esters (CEs) were extracted with hexane.The extracted CEs were analyzed by reversed phase HPLC with a UV detection at 205 nm.Cholesteryl eicosapentaenoate and docosahexaenoate ( major n-3 fatty acid cholesteryl esters) were identified by liquid chromatography-tandem mass spectrometry and cholesterol in each CE fraction was measured.Peak areas of CEs were corrected for cholesterol and CE n-3 index was calculated using the corrected peak area and expressed as the percentage of n-3 fatty acid CEs in total CEs.Results The HPLC analysis can be finished in 6 minutes.Triglycerides which interfere with the determination of n-3 fatty acid index, were hydrolyzed with ethanolic sodium hydroxide (4 mol/L) in 30 seconds.The within-run and total CVs for CE n-3 index averaged 0.66% and 0.90%, respectively.CE n-3 indexes of 70 volunteers and 36 coronary heart disease patients apparently healthy subjects and patients with coronary heart disease in Beijing Hospital appeared to be positively skewed and leptokurtic distribution ( skewness = 1.25, kurtosis = 1.70 ).The median of n-3 indices were 0.98% ( 0.37% - 2.40% ).The logarithm of n-3 index appeared to be normal distribution and the average is 0.003 7% with standard deviations of 0.15.The distribution of n-3 indices of gender groups was similar with the total.The medians of females and males were 1.08% (0.60% -2.40%) and 0.95% (0.37% -2.11%) respectively, and the former were significantly higher than the latter( t = - 3.021, P = 0.003 ).Conclusion A new method for the measurement of n-3 index of serum cholesteryl esters by HPLC has been established.It is simple and precise and can be used in predicting cardiovascular diseases risks and monitoring dietary intake of n-3 fatty acids.

Background and purpose: Cetuximab-containing regimen has been increasingly applied in the treatment of patients with metastatic colorectal cancer. Simultaneously, the predictive factors of outcome for cetuximab have been thoroughly researched. The aim of this study was to evaluate the relationship between K-ras status and efficacy of cetuximab containing regimen in the treatment of patients with metastatic eolorectal cancer. Methods: Between March 2006 and June 2008, twenty-seven patients with metastatic colorectal cancer were treated with cetuximab in combination with chemotherapy. The K-ras mutation status [wild-type (wt) or mutation (nat)] was examined by polymerase chain reaction (PCR) and direct sequencing. The influence of K-ras mutation status on efficacy of cetuximab-containing regimen was analyzed. Results: For 27 patiants in this cohort, K-ras wt was detected in 55.6% (15/27) cases and K-ras mt in 44.4% (12/27) cases. Statistically significant differences were found between the patients with K-ras wt and K-ras mt in terms of overall response rate (ORR) (66.7% vs 25.0%,P=0.035) and progression-free survival (PFS) (8 months vs 4 months, P=0.0028). However, there was no significant difference in overall survival (OS) (19 months vs 12 months, P>0.05). The most common treatment-related adverse effect was skin reaction, with incidence rate of 80.0% and 66.7% (P>0.05), respectively. No treatment related death was observed. Conclusion: K-ras mutation status is a predictive factor of good efficacy of cetuximab-containing regimen in the treatment of patients with metastatic colorectal cancer. Patients with K-ras wt could benefit from cetuximab in combination with chemotherapy.

Objective To investigate the effects of weightlessness on the apoptosis of hepatocytes in rats.Methods Eighty-four adult male Wistar rats were randomly divided into weightlessness group and control group according to the random number table.Tail-suspension was used to simulate the weightlessness model.Six rats were sacrificed in each group at the end of day 1-7.Hepatocyte apoptosis and p53 expression were detected by TUNEL assay and immunohistochemistry PV-6001,respectively.All data were analyzed by variance analysis.Results An increase of apoptosis index and up-regulated expression of p53 were detected in weightlessness group.The level of apoptotic index Was significantly higher in weightlessness group than in control group from day 1 to day 5(F=77.608,P<0.05),and the apoptotic indexes of the 2 groups were close at day 6 and day 7.The p53 proteins with positive expression were mainly detected in the nucleus.The level of positive expression of p53 in liver tissue in weightlessness group was higher than that in control group at the early stage(F=113.063,P<0.05).Few hepatocytes with positive expression were detected in control group and weightlessness group at a later stage.Conclusion There is a close relationship between hepatocyte apoptosis,changes of P53 expression and stress response and tolerance to weightlessness.

Objective To investigate the clinical significance of myasthenia gravis (MG) associated autoantibodies.Methods Titin,ryanodine receptor (RyR)and acetylcholine receptor (AChR) antibodies were examined in the sera of 74 myasthenia gravies patients by ELISA.Results AChR,Titin, RyR antibodies were detected in 77.0% (57/74),39.2% (29/74) and 32.4% (24/74) of the MG patients,respectively.For thymoma MG,AChR,Titin and RyR antibodies were detected in 76.2% (16/21),71.4% (15/21) and 52.4% (11/21) respectively.For late onset MG,Titin and RyR antibodies were detected in 77.3% (17/22) and 50.0% (11/22) respectively.With respect to the modified Osserman classification,the positve rate for Titin and RyR antibodies is much higher in more severe patients (X~2= 16.094,P=0.001;X~2=11.226,P=0.011).Titin antibodies was significantly related with RyR antibodies (r=0.380,P=0.001).Conclusions Titin and RyR antibodies show high sensitivity for thymoma MG,and the combination of serological and radiological testing can increase both sensitivity and specificity in diagnosis of thymoma MG.The levels of the two antibodies may serve as important prognosis markers in MG.The induction of the immune response against Titin and RyR and the possible pathogenic effects of the two antibodies will be further studied.