Objective To explore the association between clinical pathological characteristics and the expressions of epidermal growth factor receptor (EGFR) and protein kinase B (AKT) in gastric carcinomas.Methods The expressions of EGFR and AKT were measured with immunohistochemical method in the cancer tissues and cancer-adjacent normal tissues from 153 cases of patients with gastric cancer.The association between clinical pathological characteristics and their expressions were analyzed.Results The expressions of AKT and EGFR in gastric cancer tissues had no relationship with gender,age,pathological type,and the degree of differentiation (P ＞ 0.05).A positive correlation was existed between the EGFR and TNM stages (x2 =5.43,P ＜0.05).The AKT was positively related to the size,T stage,and TNM stage of the tumor,respectively (x2 =4.73,4.95,5.32,P ＜0.05 orP ＜0.01).The levels of AKT (x2=4.83,4.75,P ＜0.05) and EGFR(x2 =4.67,4.58,P ＜0.05) in the gastric cancer tissues with lymph node and/or distant metastasis were significantly higher than the gastric cancer tissues without metastasis,respectively.Conclusions The over-expressions of AKT and EGFR would benefit the diagnosis and stages of a gastric cancer and the determination of its metastasis.

To establish HPLC chromatographic fingerprints to control the quality of Chinese herbal medicine. In this study, fingerprints were established based on HPLC-DAD chromatographs. And with these fingerprints, content variations of three important active components catalpol, 5-hydroxymethylfurfural and acteoside in Rehmannia rhizome were analyzed during processing, as well as changes of the fingerprints. Fingerprints comparing with the standard prepared Rehmannia fingerprints which came from the mean of prepared ones randomly chosen for standard was done to seek optimal processing time. The results indicated that catalpol decreased quickly as braising prolonged and almost vanished in the end. While the active component of 5-HMF increased linearly throughout the process of braising. And the content of acteoside did not show obvious change. Similarity to standard prepared Rehmannia reached summit after braising for 26 hours. So 26 hours could be considered to be the optimum time for braising prepared Rehmannia. Chromatographic fingerprint is convenient for revealing changes of constituents and for accurately controlling quality during processing prepared Rahmannia.
Chromatography ; Chromatography, High Pressure Liquid ; methods ; Dermatoglyphics ; Drugs, Chinese Herbal ; Furaldehyde ; analogs & derivatives ; chemistry ; Glucosides ; chemistry ; Iridoids ; chemistry ; Phenols ; chemistry ; Phytotherapy ; Plant Preparations ; Plant Structures ; Rehmannia ; chemistry ; Rhizome ; chemistry

We describe a genetic transformation method of secondary somatic embryogenesis in alfalfa through cotyledon-stage somatic embryos of alfalfa infected by Agrobacterium strain GV3101. The Agrobacterium strain GV3101 contained binary vector pCAMBIA2301 that had gus gene as reporter and npt II gene as selectable marker. The infected primary embryos were induced through series of medium under 75 mg/L kanamycin selection. We obtained the transgenic alfalfa plants. Then, GUS expression in different tissue of transgenic alfalfa was tested by GUS histochemical analysis. Further, the stable integration and transformation efficiency were tested by polymerase chain reaction and Southern blotting hybridization. The result showed that GUS expression was different in different organs of transgenic alfalfa; the copy number of integrated npt II gene was from 1 to 4; the transformation efficiency via primary somatic embryogenesis was 65.82%.
Agrobacterium ; genetics ; Medicago sativa ; embryology ; genetics ; physiology ; Plant Somatic Embryogenesis Techniques ; Plants, Genetically Modified ; embryology ; genetics ; Tissue Culture Techniques ; Transformation, Genetic

OBJECTIVE: To investigate the relationship between the expression of leptin, p-mTOR protein and the pathogenesis, development and clinicopathological features in colon carcinoma. METHODS: The expression of leptin and p-mTOR protein was evaluated by immunohistochemical methods in 40 normal colon mucosas, 40 colon adenomatous polyps and 108 cases of colon carcinomas. The relationship between the staining pattern and clinicopathogical features was examined. RESULTS: The positive rates of detection of leptin in normal colon mucosa, adenomatous polyps and colon carcinomas were 10% (4/40), 27.5% (11/40), and 71.3% (77/108), respectively; with significant differences among the three groups (P<0.05). The positive rates of p-mTOR protein in the normal colon mucosa, the adenomatous polyps, and the colon carcinomas were 2.5% (1/40), 20% (8/40), and 61.1% (66/108), respectively; with significant differences among the three groups (P<0.05). The expression of leptin and p-mTOR proteins were related to invasive depth, TNM stages, lymph node metastasis, distant metastasis and tumor differentiation (P<0.05), but not to age, sex, or site (P>0.05). In colon carcinoma tissues, leptin expression was positively correlated with p-mTOR expression (P<0.01). CONCLUSION Leptin and p-mTOR proteins may play important roles in the occurrence and development of colon carcinoma. The detection of leptin and p-mTOR may be helpful for evaluation of the prognosis of the patient with colon carcinoma.
Adenocarcinoma ; metabolism ; pathology ; Adenomatous Polyps ; metabolism ; pathology ; Aged ; Colon ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Humans ; Intestinal Mucosa ; metabolism ; Leptin ; metabolism ; Male ; Middle Aged ; Neoplasm Metastasis ; Phosphorylation ; Prognosis ; TOR Serine-Threonine Kinases ; metabolism

Background and purpose:It is still a great challenge to clarify the signal molecules that mediate the communication between cancer-associated fibroblasts(CAFs)and tumor cells.These signal molecules are very important for cancer metastasis.The purpose of this study was to explore the communication mechanism of pleckstrin-2/miR-196a signal axis mediated by lung cancer cells in tumor microenvironment.Methods:Human lung adenocarcinoma cell line H1299 and human embryonic lung cell MRC-5 were selected as the research objects.H1299 cells were transfected with lentivirus(PLEK2)expressing PLEK2 and Vector control,and exosomes(Vector_exo,PLEK2_exo)were isolated after 24 h of transfection.MRC-5 cells were transfected with miR-196a mimetic or inhibitor.The expressions of PLEK2 and epithelial-mesenchymal transition(EMT)-related proteins were analyzed by Western blot.The expression of miR-196a was analyzed by polymerase chain reaction(PCR),and the metastasis and invasion ability of cells were determined by transwell assay.Six female BALB/c-nu mice were randomly divided into Vector group and PLEK2 group,with 3 mice in each group.Mice in each group were injected with H1299 cells transfected with Vector or PLEK2 through the tail vein.After 4 weeks,lung tissue was taken out for H-E staining and immunohistochemical staining to analyze the expression of α-smooth muscle actin(α-SMA).All animal experiments were approved by the ethics committee of First Hospital of Changsha City(Changsha Hospital,Xiangya School of Medicine,Central South University)(ethics number:EI-2021-103).Results:Compared with the Vector group,the number of pulmonary metastatic nodules and the expression of α-SMA in metastatic cancer in PLEK2 group increased significantly(P＜0.001).Compared with Vector group,the expression level of miR-196a in H1229 cells in PLEK2 group increased significantly(P＜0.05),and the expression level of miR-196a was significantly higher in PLEK2_exo than in Vector_exo(P＜0.05).Compared with Vector_exo group,the expression levels of miR-196a,α-SMA and fibroblast activation protein(FAP)in MRC-5 cells in PLEK2_exo group increased significantly(P＜0.05).Compared with the negative control(NC),the expression levels of α-SMA and FAP in MRC-5 cells transfected with miR-196a increased significantly(P＜0.05).On the contrary,by transfection with miR-196a inhibitors(si-miR-196a#1 and si-miR-196a#),the expression levels of α-SMA and FAP were significantly inhibited(P＜0.05).Compared with NC-CM group,the number of metastatic cells,invasive cells and the expression of vimentin in miR-196a-CM group increased significantly(P＜0.001),and the expression of E-cadherin decreased significantly(P＜0.001).In addition,compared with Vector_exo-CM group,PLEK2_exo-CM group had significant increase in number of metastatic and invasive cells and the expression of vimentin(P＜0.01),and significant decrease in the expression of E-cadherin(P＜0.001).Conclusion:Upregulation of PLEK2 can enhance the level of exosomes miR-196a derived from lung cancer cells,thereby promoting the activation of CAFs.The activated CAFs can further enhance the invasive ability of lung cancer cells.