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Objective To explore the diagnostic value of cytologic examination of cerebrospinal fluid (CSF) combined with cell immunochemical staining examination in the patients with meningeal carcinomatosis in the clinical track.Methods CSF was centrifuged and precipitated using the StatSpin Cytofuge 12 centrifuge. It was dyed by the May-Grunwald-Giemsa Staining (MGG) method and cell immunochemical staining (S-P method). Clinical data of 16 cases with the diagnosis of meningeal carcinomatosis were retrospectively analyzed. Results Sixteen patients diagnosed meningeal carcinomatosis prior to imaging findings in 11 of 16 patients were diagnosed meningeal carcinomatosis through the cerebrospinal fluid cytology combined with cell immunochemical staining prior to imaging findings. CSF cytology observed the cancer cell pleomorphism. Cell immunochemical staining examination clearly distinguished classification organic source of meningeal carcinomatosis. Conclusions CSF cytology combined with cell immunochemical staining examination is one of the important means in tracking intracranial metastatic tumor in clinic.
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Objective In vitro isolation of adenohypophyseal mainly includes enzymatic digestion and mechanical methods.But there is no relevant report about which method is better.In this paper,several mainstream methods of the vitro isolation of adenohypophysis cells in rats are compared and the identification of gonadotropin secretory cells is carried out to figure out which preparation method is more convenient and efficient.Methods 48 mature female SD rats were randomly divided into 4 groups:trypsin group (0.25％ trypsin EDTA digestion),Ⅳ-type collagen enzyme group (Ⅳ type collagen enzyme digestion),mechanical separation group (200 mesh cell sieves grinding) and trypsin digestion plus mechanical separation group (0.25％ trypsin EDTA digestion plus 200 mesh cell sieves grinding).12 rats in each group.The effect of these 4 methods was evaluated.Secondly,the primary cells of each group were cultured.We dynamically observed the vitro growth of cells in each group.Finally,the cells were identified by using the immunocytochemi-cal staining technique Results Compared with the mechanical separation group and-ⅣV type collagen enzyme group [(90.2 ± 0.96) ％,(93.32± 1.77)％],Cell viability of trypsin group and trypsin digestion plus mechanical separation group was elevated [(94.11 ± 1.71) ％,(94.92± 1.92) ％] (P＜0.05).Morphological observation:The pituitary ceils prepared by each methods were all round with strong refraction and clear edges and began to partly adhere to the wall after cultured for 16-24 hours.Then most of the cells adhered to wall after 48-72 hours,while the early glandular ceils were still round.But the volume was smaller than before.Also,a small amount of large cells were scattered among them.7 days later,the cells began to become polygonal or spindle shaped and were connected to pieces.Till around 10 days,the fibrosis gradually became obvious;Immunocytochemical staining:It is indicated that FSH positive cells were larger in volume while less in number and scattered in the cytoplasm.And the positive products,showing blue,were located in the cytoplasm;The Enzyme linked immunosorbent assay (ELISA) was used to determine the supematant of cells and confirmed that the cells in vitro still had hormone secretion functions.Conclusion Compared with the single cell preparation method,the trypsin digestion plus mechanical separation method has certain advantages;The anterior pituitary cells vitro cultured are in good health conditions and still have secrete functions.They are tested to be competent for related scientific experiments.
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Objective To study the change of special antibodies titer IgG, IgM and nucleocaspid to SARS coronavirus (CoV) and observing the expression of stomach and enteric involvement on SARS-CoV infection by monoclonal antibody against N protein of SARS-CoV in the 7- year recovery period among family clustering cases of severe acute respiratory syndrome. Methods Special antibody titer to SARS-CoV of 14 patients from 5 different families and their 10 kinfolks continuously tested by IFA and antigen-capturing ELISA methods. Samples were taken in the 1st-7th year periods after SARS patients infected by SARS-CoV, being diluted and measured on it titers of three kinds of antibodies. Immunochemical staining with monoclonal antibody (mAb) against N protein of SARS-CoV was used to determine the stomach and enteric tissues among 5 SARS patients with their nucleocaspid antibody titer ascended obviously after 1st-7th year. Results When testing the IgG antibody titer of the 14 SARS patients by IFA method, the average titer was 1/71 (95%CI:1/58-1/85) in the 1st year, but began to descend in the following years, and the IgG antibody of the most SARS patients disappeared in the 7th year. Regarding the IgM titer, it disappeared in most of the SARS patients 1 year later. The average value of nucleocaspid antibody titer was 1/146 (95% CI:1/122-1/171) in the 1st year, and it descended as the IgG antibody titer did. In 5 cases, differences appeared.The nucleocaspid antibody titer was between 1/156 and 1/210 in 3 cases, and 2 cases were normal.Immunochemical staining with mAb against N protein of SARS-CoV was identified in the stomach and enteric tissues of 5 SARS patients with the nucleocaspid antibody titer increased significantly, 1st-7th year later. The five patients were detected by gastroscopy detection and cell immunohistochemistry test. 3 cases showed N protein antibody positive in the serum, and positive immunohistochemical expression in most of the cytoplasm in the gastric tissue mucous gland epithelial cells. 1 case also expressed in the intestinal tissue slurry columnar epithelium and interstitial cells. The other two cases showed negative on both serum N protein antibody and immunohistochemical expression. The biopsy results of the 5 patients were as follows: 1 case diagnosed as "signet-ring cell carcinoma of the stomach and rectum multiple transfer", 1 case of gastric polyp, 1 case of superficial antral gastritis and 2 cases were normal. Conclusion By testing the special IgG, IgM, nucleocaspid antibody to SARS-CoV of the 14 family clustering cases , we found that they all decreased in the 7th year, and most of them disappeared. The nucleocaspid antibody titer was related to pathogenetic condition. SARS-CoV was proved to be still present in stomach and enteric tissues of SARS patients with the nucleocaspid antibody titer increased significantly after the 7th year.