Objective: To reconstruct a substractive complementary deoxyribornucleic acid (cDNA) library of genes sensitive to mechanical stretch in human osteoblast like cells.Methods: Mechanical stretch at 12 cycles per minute was applied to human osteoblast like cells Saos-2 and the deformation of the stretched cells was 12%. Twelve hours after loading, mRNAs were isolated from both stretched and unstretched cells. Substractive cDNA library of the genes sensitive to stretch was constructed with the technique based on polymerase chain reaction (PCR) and substractive hybridization. Primary sequencing of clones in the library was carried out. Results: A substractive cDNA library of genes sensitive to stretch was constructed with a capacity of about 200 clones. According to the results of sequencing, most genes in the library were related to the mechanical stimulation. One novel gene fragment was obtained. Conclusion: The method used in the experiment is effective in cloning genes sensitive to mechanical stretch.

Objective:To identify the function of the 5′-flank upstream regulation region of human CD226 gene.Methods:The upstream regulation region of CD226 was cloned by PCR and ligated into pGL3 vector. Then the vector was transfected into Jurkat cell and luciferase activity was detected after 48 h culture.Results:CD226 gene may have two promoters, P1 and P2,which were located at the region of -843--319 bp and +1-+181 bp respectively, and PMA can up-regulate P1 while down-regulate P2. Both P1 and P2 can be up-regulated by A23187, especially P2.Conclusion:CD226 gene may have two promoters, and PMA and A23187 can regulate CD226 promoter activity in the similar pattern of protein level.

Objective:To compare the immunogenity of the AST artificial antigen with two different carrier proteins, bovine serum albumin(BSA) and keyhole limpet hemocyanin(KLH).Methods: The hapten Astragaloside Ⅳ(AST) was linked to carrier protein BSA or KLH as immunogen,and linked ovalbumin(OVA) as coating antigen by the sodium periodate method.Then the 6-8 weeks BALB/c mice were immunized with AST-BSA or AST-KLH.The titer and specificity of the antiserum were detected by indirect enzyme-linked immunosorbent assay ( iELISA ) and indirect competitive enzyme-linked immunosorbent assay ( icELISA ) respectively.Results:The antiserum of the mice immunized with AST-KLH had better specificity than which immunized with AST-BSA.The IC50 of AST-KLH antiserum was about 5 μg/ml in the detection by icELISA.The high titer of both antiserums was about 1∶51 200 after immunizing mouse.Conclusion:The conjugate of AST-KLH was testified to have a better immunogenity in comparison with AST-BSA,this investigation would be the foundation of the further research of AST immunoassay.

Objective:To investigate the expression and function of CD226 on NK cell clone Methods:NK cell clone was established by limited dilution, and identified by FCM The function of CD226 on the cytotoxicity of NK cell clone was detected by RCA and the NK cell clone secreted cytokines in the supernatants during the killing phase were measured by ELISA Results:One NK cell clone was obtained by limited dilution The cytotoxicity of this clone was upregulated markedly by CD226 mAb, and the secretion levels of IFN ? and GM CSF by NK cell clone increased obviously in RCA Conclusion:CD226 is a novel NK cell activation receptor, and the elevated IFN ? and GM CSF levels may be related to CD226 mAb enhanced NK function

Objective:To clone mouse CD226(platelet and T cell activation antigen1,PTA1).Methods:Specific primers were designed and sythesized according to the EST sequence from GenBank,which had 51% homology to human CD226 on the amino acid level.And then,the cDNA of mouse CD226 was cloned from the thymus of 4 week old BALB/c mouse by using RACE technique.Results:The length of mouse CD226 cDNA is 2 223 bp,with the open reading frame(ORF) of 1 002 bp,encoding 333 amino acids,which is 3 amino acids shorter than its counterpart in human.The mouse CD226 belongs to IgSF,and shares 53% homology with human PTA1 on amino acid level.Besides,three isoforms of mouse PTA1 were also cloned at the mean time.Conclusion:The molecular cloning of mouse PTA1 lays the foundation for the in vivo studies on the biological function of this molecule,as well as the studies of this molecule in gene knockout mouse and transgenic mouse.

OBJECTIVE: To investigate the significance and relationship between the expression of FOXC1 and clinicopathological features, and to explore its correlation with E-cadherin. METHOD: Immunohistochemical SP method was used to detected the expression of FOXC1 in nasopharyngeal carcinoma tissues and nasopharyngitis tissues. RESULT: (1) Immunoreaction to FOXC1 was mainly located in nucleus of nasopharyngeal carcinoma cells. The positive expression rate of FOXC1 in nasopharyngeal carcinoma tissues was 85.3% (81/95), which was significantly higher than that in nasopharyngitis tissues (59.4%) (P < 0.05). (2) The expression of FOXC1 was not related to patients' age and gender, clinical stage of cancer and lymph node metastasis (P > 0.05). (3) There was a correlation between the expression of FOXC1 and down-regulated expression of E-cadherin in nasopharyngeal carcinoma tissues (P < 0.05). CONCLUSION FOXC1 may play an important role in generation and progression of nasopharyngeal carcinoma, there may be a correlation between the expression of FOXC1 and down-regulated expression of E-cadherin, also FOXC1 may play an important role in the process of EMT in nasopharyngeal carcinoma by regulating E-cadherin.
Adolescent ; Adult ; Aged ; Antigens, CD ; Cadherins ; metabolism ; Carcinoma ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Nasopharyngitis ; metabolism ; Young Adult

Objective:To investigate the risk factors for early complications after laparoscopy-assisted gastrectomy in patients with gastric cancer.Methods:The retrospective case-control study was conducted. The clinicopathological data of 196 patients who underwent laparos-copy-assisted radical gastrectomy at Peking Union Medical College Hospital from March 2016 to March 2019 were collected. There were 144 males and 52 females, aged (61±10)years. Observation indicators: (1) early complications after laparoscopy-assisted radical gastrectomy and treatment; (2) analysis of risk factors for early complications after laparoscopy-assisted radical gastrectomy.Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( P25,P75). Count data were represented as absolute numbers. Univariate analysis was conducted using the t test, Mann-Whitney U test or chi-square test. Multivariate analysis was conducted using the Logistic regressional model. Results:(1) Early complications after laparoscopy-assisted radical gastrectomy and treatment: 51 of 196 patients had early postoperative complications, including 7 cases of grade Ⅰ according to Clavien-Dindo classi-fication system, 32 cases of grade Ⅱ, 9 cases of grade Ⅲa, 3 cases of grade Ⅲb. There was no grade Ⅳ or Ⅴ complication. There were 25 cases with abdominal complications, 7 cases with thoracic complications, 3 cases with internal/catheter related complications and 16 cases with other unclassified complications. All patients with complications were improved after symptomatic and supportive treatments. (2) Analysis of risk factors for early complications after laparoscopy-assisted radical gastrectomy: results of univariate analysis showed that the lymphocyte count, neutrophil-to-lymphocyte ratio, radiotherapy, operation time, volume of intraoperative blood loss, T stage, lymph node metastasis were related factors for early complications after laparoscopy-assisted radical gastrectomy in patients with gastric cancer ( Z=?2.048, χ2=6.385, 4.168, 8.068, 6.336, 12.497, 7.522, P<0.05). Results of multivariate analysis showed that the neutrophil/lymphocyte ratio ≥1.96, operation time ≥222 minutes, and lymph node metastasis were independent risk factors for early complica-tions after laparoscopy-assisted radical gastrectomy in patients with gastric cancer ( odds ratio=2.279, 2.245, 2.226, 95% confidence interval as 1.149-4.519, 1.116-4.517, 1.125-4.402, P<0.05). Conclusions:The abdominal complications are the most common early complications after laparoscopy-assisted radical gastrectomy. The neutrophil-to-lymphocyte ratio ≥1.96, operation time ≥222 minutes, and lymph node metastasis are independent risk factors for early complications after laparoscopy-assisted radical gastrectomy in patients with gastric cancer.

Using the liquid chromatography-tandem mass spectrometry (LC-MS/MS) to determine the plasma level of Lyso-GL3 in patients with Fabry disease and to analyze the clinical application of the method.