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 study the influence of transferring a dominant-negative Stat3 gene, Stat3?on colon cancer cells' proliferation and apoptosis in vitro. Methods Cell culture of human colon cancer cell line SW480 and transient transfection were used to evaluate the effect of transferring Stat3?to cancer cells. Cell proliferation, cell cycle and apoptosis were quantified by MTT and flow cytometry, respectively. The mRNA expression of Stat3's target gene cyclin D1 and bcl-xL was detected by reverse transcription polymerase chain reaction. Independent t tests were used for data statistics. Results 36 h after Stat3?plasmids transfection, proliferation of SW480 cells was significantly inhibited (t =5. 216,P = 0.006); cell proportion of G0/G1 phase increased from 40.37% to 67.25% and early apoptosis cells increased from 5. 34% to 24. 42% ; mRNA expression of cyclin Dl and bcl-xL declined significantly (t = 5.288,P=0.010;t=3.517,P=0.025). Conclusion Blocking Stat3 signaling pathway by transfection of Stat3?plasmid inhibits the proliferation and promotes apoptosis of colon cancer cells, which provides a experimental foundation of Stat3 targeted colon cancer gene therapy.

Objective: To observe the effects of needles with different diameters on the gastrointestinal function in mice. Methods: Eighteen Kunming mice were randomly divided into group of 0.25 mm needle, group of 0.35 mm needle, and control group. The acupoint of Zusanli (ST 36) was needled once a day for 5 days. The effects of needles with different diameters were observed by measuring the distance of the carbon moved in the intestine. Results: The distance of the carbon moved in the intestine was longer in the acupuncture group than in the control group (P<0.05), and it was longer in the group of 0.35 mm needle than in the group of 0.30 mm needle, there was no significant difference (P>0.05). Conclusion: Acupuncture treatment can enhance the peristalsis function of stomach and intestine in mice. The diameter of needle has no effect on the gastrointestinal function.

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.

Aim To observe the improvement of visual-auditory cognitive functions in the human entering high altitude by taking in tea polyphenols.Methods Thirty eight males living at 3 700 m high altitudes for 90 days constantly were randomly divided into two groups: ①group Ⅰ(placebo,40 mg/day); ②group Ⅱ(TP,300 mg/day).Cognitive functions were measured by integrated visual and auditory continuous performance test and the difference between groups was evaluated by the comparisons of post-treatment to pre-treatment.Results Compared with pre-treatment,PruA was significantly higher after taking in TP(P