Objective To investigate the clinical manifestations,pathological diagnosis,treatment meth-od and prognosis of primary gastric adenosquamous carcinoma.Methods The clinical data of 20 patients with primary gastric adenosquamous carcinoma receiving the treatment in this hospital from January 1,2013 to No-vember 30,2022 were retrospectively analyzed,and their clinicopathological characteristics and prognosis were analyzed.Results Among the 20 patients,14 cases were male and 6 cases were female,with a median age of 59(36,74)years old.Upper abdominal discomfort,pain and weight loss were the main clinical symptoms.Ser-um carbohydrate antigen 19-9(CA19-9)levels were elevated in some patients.The image examination indica-ted the gastric occupation.Among 20 cases,the tumors were most common in the lower third of the stomach(14 cases),followed by the upper third(1 case)and the middle third(5 cases).The most common tumors were in the lower one-third(14 cases)of the stomach,followed by the middle one-third(5 cases)and upper one-third(1 case).In histomorphology,the gastric tumor cells contained two components,adenocarcinoma and squamous cell carcinoma,and squamous cell carcinoma accounted for more than 25％of tumor cells.Immuno-histochemistry showed that the partial tumor cells of adenocarcinoma expressed CK8/18 and partial tumor cells of squamous cell carcinoma expressed p40.All 20 cases performed the surgical treatment.Only 6 cases survived and the others died of tumor recurrence or metastasis.The adenosquamous carcinoma proportion and Ki-67 were correlated with the prognosis in the patients with gastric adenosquamous carcinoma(P＜0.05).The survival curve constructed by the Kaplan-Meier method showed that when the proportion of squamous carcinoma was more than 35％,the prognosis of the patients was good.Conclusion Primary gastric adenosquamous carcinoma,mainly composed of adenocarcinoma,may be correlated with a higher risk of metastasis.

Objective To investigate antigen optimization of Shisa like protein 1 (SHISAL1) for preparing mouse anti-human SHISAL1 polyclonal antibody and to identify the specificity of the prepared antibody. Methods Bioinformatics was employed to predict the antigenic epitope region of SHISAL1 protein, and then a polypeptide composed of amino acid residues from the site of 28 to 97 of SHISAL1, termed SHISAL1-N, was selected as the antigen. The coding region of SHISAL1-N was cloned by molecular cloning technique, and then it was inserted into pET-28a to generate pET28a-SHISAL1-N recombinant plasmid. The two recombinant plasmids pET28a-SHISAL1-N and pET28a-SHISAL1 were transformed into BL21 (DE3) bacteria and induced to express by IPTG. The two proteins were purified and immunized to female Kunming mice, respectively. The specificities and sensitivities of the acquired antibodies were detected by Western blot analysis, immunoprecipitation and immunofluorescent cytochemical staining. Results pET28a-SHISAL1-N recombinant plasmid was successfully constructed, and the two fused proteins, SHISAL1 and SHISAL1-N, were induced to express. Moreover, two types of SHISAL1 mouse polyclonal antibodies, derived from SHISAL1-N and SHISAL1 antigens, were obtained. Western blot results showed that the antibody prepared from SHISAL1 antigen was less specific and sensitive compared with the antibody prepared from SHISAL1-N antigen which could specifically identify different endogenous SHISAL1 protein. Immunoprecipitation results showed that SHISAL1-N antibody could specifically pull down SHIISAL1 protein in hepatocellular carcinoma cells and immunofluorescence results demonstrated that SHISAL1-N antibody could specifically bind to SHISAL1 protein in the cytoplasm. Conclusion We have optimized the SHISAL1 antigen and prepared the mouse anti-human SHISAL1 polyclonal antibodies successfully, which can be used for Western blot analysis, immunoprecipitation and immunofluorescence cytochemical staining.
Animals ; Female ; Humans ; Mice ; Antibodies ; Antibody Specificity ; Blotting, Western ; Cloning, Molecular ; Epitopes/genetics*