Background and purpose:To study the expression of PTEN and P53 and analyse their roles in HGC oncogenesis,and to discover a good marker for diagnosis and prognosis of HGC.Methods:The expression of PTEN mRNA,PTEN protein and P53 protein were detected by in situ hybridization(DNA-RNA) and immunohistochemistry(SP method) in 64 cases of HGC.We took 22 cases of gallbladder adenoma and 10 cases of chronic cholecystitis as controls.Results:① The positive expression rates of PTEN mRNA and protein in HGC(56.3%,59.4%) were significantly lower than that in gallbladder adenoma(81.8%,90.8%) and chronic cholecystitis(100%,100%)(P

Objective To establish differential protein expressing profiles of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by two-dimensional differential gel electrophorosis (2-D DIGE) so as to investigate the metastatic molecular mechanism of the gastric cancer.Methods After obtaining 6 samples of human primary gastric cancer and metastatic lymph node tissues,with manual no-staining frozen sections microdissection,human gastric adenocarcinoma cells from primary or metastatic lymph node tissues were isolated,and then the total proteins were extracted and purified.Highly sensitive 2-D DIGE was used to separate the total protein differentially expressed in the cells.The proteins were visualized by using a fluorescence scanner at appropriate wavelengths for Cy2,Cy3 and Cy5 dyes (Typhoon 9400).Image analysis was carried out with the DeCyder-Differential analysis software (Biological Variation Analysis version 5.0).Results Not only can the study procure defined adenocarcinoma cell populations from gastric primary or metastatic lymph node tissues,but also can resolve the problem of the change in 2-D DIGE patterns because of the varying in protein changes owing to dyeing.All these showed that the technique was simple,easy to perform,versatile and of particular usefulness when laser capture microdissection (LCM) was practically unavailable.The 2-D DIGE patterns with high resolution and reproducibility from adenocarcinoma cells in gastric primary or metastatic lymph node tissues were obtained.The number of spots in Gel1,Gel2 were 1 416 (similar 1 062,decrease 277,increase 77),1 299 (similar 1 050,decrease 157,increase 92),respectively.A total of 11 differential proteins were acquired by image analysis with DeCyder-Differential analysis software (Biological Variation Analysis version 5.0).Conclusions In this report,a simple,easy to proform method of protein epuration,manual no-staining frozen sections microdissection is described,and have used the highly sensitive 2-D DIGE for the identification of proteins differentially expressing in human gastric adenocarcinoma cells from primary or metastatic lymph node tissues.These results provide a fundamental basis for further study of metastatic mechanism of gastric cancer and screen its specific markers.

Objective To identify the differentially-expressed proteins of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by comparative proteomics technology, and to screen the specific metastatic-associated proteins so as to investigate the metastatic molecular mechanism of lymph node metastasis in gastric cancer. Methods 11differential proteins were acquired previously from primary and metastatic lymphnode tissues in gastric adenocarcinoma patients by 2D-DIGE. Some selected differential protein spots were identified by PMF based on MALDI-TOF-MS and database search. Immunohistochemical staining of HSP70 was used to evaluate the reliability of the proteomic analysis results. Results After analyzed on 11 differential proteins by in-gel trypsin digestion and MALDI-TOF-MS-based PMF analysis, a total of 5 differential proteins were identified by searching Mascot-database, HSP70ˊs 8 isoform 2 variant, chaperonin, chaperonin, leucine aminopeptidase, predicted: hypothetical protein XP_515584. Among the differential proteins identified, the levels of HSP70, chaperonin, leucine aminopeptidase expression had a significant up-regulation in gastric primary cancer compared with metastatic lymph node. HSP70 expression rate increased with the metastasis of lymph node and the progress of gastric cancer, agreed with the proteomics results. Conclusions They are similar in differentially-expressed proteins in primary or metastatic lymph node tissues because of the uniformity in source and differention. There are few protein changes in cancer cells between them, taking part in the metastatic of gastric cancer. HSP70 takes part in the progress of gastric cancer and relates to the metastasis of lymph node and malignant degree.

BACKGROUND:It is particularly important to establish an ideal animal model of pulmonary fibrosis to investigate the underlying pathogenesis and screen effective drugs to prevent and control pulmonary fibrosis. OBJECTIVE: To establish a modified scheme of establishing mouse models that can reflect pulmonary fibrosis formation in humans. METHODS: Fifty-six male C57BL/6 mice were randomly divided into two groups: A (a single large-dose injection) and B (multiple smal-dose injections). Mice in group A were subjected to a single intravenous injection of bleomycin 200 mg/kgviathe tail vein; and mice in group B received intravenous injections of bleomycin 50 mg/kg via the tail vein per week, totaly for 6 weeks. 

BACKGROUND:There is no effective drug for idiopathic pulmonary fibrosis (IPF), because of a lack of the animal model imitating the complete pathogenesis of human IPF. Therefore, it is critical to establish an ideal animal IPF model used for investigating the underlying pathogenesis and developing a kind of effective drug. OBJECTIVE:To establish an animal model that can mimic more characters of human IPF. METHODS:Seventy male C57BL/6 mice were randomly divided into two groups, fol owed by subjected to the intraperitoneal injection of bleomycin (35 mg/kg) on days 1, 4, 8, 11, 15, 18, 22, and 25, twice (group A) or once (group B) a week. Mice were sacrificed at 2, 4, 6, 8, and 10 weeks after the eighth injection, and the lung tissues were moved used for hematoxylin-eosin, Masson and immunohistochemical stainings. RESULTS AND CONCLUSION:There were various degrees of alveolitis and pulmonary fibrosis in the two groups at different time points after the last injection. The scores of alveolitis and pulmonary fibrosis in the group A began to gradual y increase from the 2nd week and reached the highest level at the 6th-8th weeks until the 10th week. In contrast, the scores of alveolitis and pulmonary fibrosis in the group B peaked at the 2nd week, then fluctuately decreased, and were significantly lower than those in the group A at the 6th week (P<0.05). Immunohistochemistry showed that type I col agen deposition was mainly distributed in the subpleural region, peri-vascular region and alveolar septa, which was consistent with Masson staining findings. The expression levels of transforming growth factorβ1 (TGF-β1) andα-smooth muscle actin (α-SMA) in the regions developing alveolitis and pulmonary fibrosis were significantly increased. In the group A, the expression levels of type I col agen, TGF-β1,α-SMA, and the hydroxyproline content in the lung tissues reached the peak level at 6-8 weeks. However, in the group B, al above indicators reached the highest level at the 2nd week, but gradual y decreased thereafter. At the 4th week, the expression Levels of TGF-β1 andα-SMA in the group B were significantly lower than those in the group A (P<0.05). At the 6th week, the hydroxyproline and type I col agen levels in the group B were significantly lower than those in the group A (P<0.05). In conclusion, the mouse model of pulmonary fibrosis induced by intraperitoneal injection of 35 mg/kg bleomycin twice weekly can be used to mimic the repetitive wound healing process, pathological morphology and cytokine changes of human IPF, which is prone to administration, with better stability and repeatability. This model is of great significance for the study on IPF. Subject headings:Disease Models, Animal;Pulmonary Fibrosis;Bleomycin

OBJECTIVE: To evaluate the cytotoxicity of a new type of titanium alloy Ti-25Nb-10Ta-1Zr-0.2Fe by studying the induced proliferation of L929 cells in contrast with other titania widely used in clinical practice. METHODS: The cell line was treated with extracting liquid containing different concentrations of titanium alloys. The number and morphology of cells was observed under an inverted phase contrast microscope. MTT was used to measure the relative growth rate (RGR) and judge the cytotoxicity grade. Flow cytometry was used to observe cell cycle progression. RESULTS: The RGR of TNTZ group cells at the 3 time points was (93.7±0.8), (100.6±0.4), and (106.4±0.3); the cytotoxicity grade was 1, 0 and 0 after treating for 1, 3 and 5 days; with influence on neither the cell morphology nor the cell cycle. The flow cytometry showed that the sequence of S phase cells was Ti>TNTZ>TC4>blank control >TC4ELI, with no significant difference (P>0.05). None of the 4 materials inhibited the cell proliferation. CONCLUSION The cell morphology and proliferation are not affected by TNTZ. The new titaniu alloys shows good cyto-compatibility. The cytotoxicity is grade 0, meeting the clinical application standard.
Alloys ; toxicity ; Animals ; Cell Line ; Dental Alloys ; toxicity ; Fibroblasts ; cytology ; Mice ; Niobium ; Tantalum ; Titanium ; toxicity ; Toxicity Tests ; Zirconium

Objective To study the clinical effect of continuous lumbar cistern using precision infusion set control and drainage of intracranial infection after craniotomy with Incision healing bad cerebrospinal fluid leakage. Methods From October 2008 to October 2013, 50 cases of postoperative intracranial infection and cerebrospinal fluid leakage patients using continuous lumbar cistern with precision infusion set control cerebrospinal fluid drainage were retrospectively analyzed. Results These 50 patients,after traumatic brain injury after decompressive craniectomy with the poor wound healing 20 cases of cerebrospinal fluid leakage , decompressive craniectomy in hypertensive intracerebral hemorrhage with hydrops under skin flap with 20 cases of cerebrospinal fluid leakage ,all patients recovered and were discharged from the hospital. Conclusion Intracranial infection and cerebrospinal fluid leakage using continuous lumbar cistern with precision infusion set drainage of cerebrospinal fluid ,with systemic application of antibiotics to treatment of post-operative cerebrospinal fluid leakage operation incision heali is a method for safety , good intracranial infection.

OBJECTIVE: To compare 3 commonly used methods for drug delivery via the lumbar spinal subarachnoid space in rats. METHODS: We compared the effects of 3 methods for drug delivery via the lumbar spinal subarachnoid space in Sprague Dawley rats, namely acute needle puncture, chronic catheterization via laminectomy, and non-laminectomized catheterization. Body weight changes of the rats were measured, and their general and neurological conditions were assessed after the surgeries. The motor function of the rats was examined using rota rod test both before and after the surgeries. Nociceptive tests were performed to assess nociception of the rats. HE staining was used to examine local inflammation caused by the surgeries in the lumbar spinal cord tissue, and lidocaine paralysis detection and toluidine blue dye assay were used to confirm the precision of drug delivery using the 3 methods. RESULTS: Both needle puncture and catheterization via laminectomy resulted in a relatively low success rate of surgery and caused neurological abnormalities, severe motor dysfunction, hyperalgesia, allodynia and local inflammation. Catheterization without laminectomy had the highest success rate of surgery, and induced only mild agitation, slight cerebral spinal fluid leakage, mild sensory and motor abnormalities, and minimum pathology in the lumbar spinal cord. Catheterization without laminectomy produced less detectable effects on the behaviors in the rats and was well tolerated compared to the other two methods with also higher precision of drug delivery. CONCLUSIONS Catheterization without laminectomy is a safe, accurate and effective approach to lumbar drug delivery in rats.