KDELR2 is an unfavorable prognostic biomarker and regulates CCND1 to promote tumor progression in glioma.

BACKGROUND: The KDEL receptor is a seven-transmembrane-domain protein, which plays a key role in ER quality control and in the ER stress response, KDELR2 involved in regulation of cellular functions, including cell proliferation, survival, promotes glioblastoma tumorigenesis. The aim of this study was to investigate the clinicpathological value and biological role of KDELR2 in glioma. METHODS: We studied the expression of KEDLR2 and its association with the prognosis through the TCGA, CGGA, and GSE16011 database. To explore the role of KDELR2 in glioma, KDELR2 siRNA was constructed and transfected into U87 glioma cells. CCK-8, colony formation and Transwell assays were used to investigate the roles of KDELR2 on GBM cell proliferation. We further studied the effect of KDELR2 on tumorigenesis in animal model. Additionally, flow cytometry was used to monitor the changes in the cell cycle and apoptosis following transfection with KDELR2 siRNA. We applied GeneChip primeview expression array to analysis the differential gene expression profiling. Ingenuity Pathway Analysis to show that KDELR2 has a significant impact in canonical pathway in cell cycle regulation and participate in multiple pathways. And we detected the cell cycle proteins CCND1 expression by Western blot analysis. RESULTS: Our results showed that KDELR2 was up-regulated in glioma tissue and cell lines. Knockdown KDELR2 was able to reduce cell viability, promote cell cycle arrest at the G1 phase, and induce apoptotic cell death. Moreover, our results suggested that KDELR2 regulated the cellular functions of U87 cells by targeting CCND1. Therefore, we demonstrated that KDELR2 is a novel biomarker in glioma. CONCLUSIONS: KDELR2 is highly expressed in human glioma tissues and cell lines, a higher expression of KDELR2 is associated with a poor prognosis of glioma patients. Moreover, KDELR2 regulated the cellular functions of U87 cells by targeting CCND1. The KDELR2/CCND1 axis may provide a new therapeutic target for the treatment of glioma and deepen our understanding of glioma mechanisms.

Mechanism of Liushenwan and Realgar in Inducing Apoptosis and DNA Damage in Human Endometrial Cancer JEC Cells / 中国实验方剂学杂志

Objective:Compare the anti-tumor effect and mechanism of Liushenwan and realgar (As4S4) on human endometrial cancer cells JEC. Method:The release of As in Liushenwan and As4S4 was measured by atomic absorption spectrometry. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used for cell proliferation, and cell migration was measured by Transwell assay. Flow cytometry and Western Blot were used to determine apoptosis and DNA damage. Result:The dissolution of As in Liushenwan was 17.4%, and that of As4S4 was only 1.6% according to atomic absorption assay. With the same content of As, compared with the As4S4 group, the cell viability in the 3，10 mg·L-1 Liushenwan groups was decreased (P<0.05), the early apoptosis rate was significantly increased in 0.25，0.5，1 mg·L-1 Liushenwan groups (P<0.05), the rate of cell migration was decreased in 1 mg·L-1 Liushenwan group (P<0.05), the expressions of cleaved cysteinyl aspartate-specific proteinase-3 (cleaved Caspase-3), cleaved cysteinyl aspartate-specific proteinase-7 (cleaved Caspase-7), cleaved poly ADP-ribose polymerase (cleaved PARP), phosphorylated histone (p-H2AX), phosphorylation of checkpoint kinase 2 (p-CHK2) and ataxia-telangiectasia mutated (ATM) were increased in 1 mg·L-1 Liushenwan group (P<0.05), while the expression of phosphorylation of ataxia-telangiectasia mutated rad3-related (ATR) was decreased in 1 mg·L-1 Liushenwan group (P<0.05), with no significant changes in the expressions of cysteinyl aspartate-specific proteinase-3 (Caspase-3) and cysteinyl aspartate-specific proteinase-7 (Caspase-7). Conclusion:With the same content of As, both Liushenwan and As4S4 could inhibit JEC cell proliferation and migration, and induce cell apoptosis and DNA damage. Liushenwan has a stronger effect than As4S4. It is suggested that there are other components in Liushenwan with an anti-tumor effect in cooperation with As.

Determination of effective components and analysis of antioxidant enzyme activities of Ampelopsis grossedentata under different breeding modes / 中成药

AIM To determine effective components of Ampelopsis grossedentata (Hand.-Mazz.) W.T.Wang under different breeding modes and to analyze the antioxidant enzyme activities.METHODS Single factor and orthogonal tests were adopted in screening the optimal extraction process of antioxidant enzymes and analyzing antioxidant enzyme activities of A.grossedentata under common cutting,two-phase cutting and tissue culture.Solvent extraction method was adopted in extracting the effective components of three breeding modes in stem leaves of A.grossedentata,whose contents were determined.RESULTS In three breeding modes,the content of polysaccharides in tissue culture seedling was the highest,while the contents of flavonoids and polyphenols in the two stage cutting seedlings were the highest.The optimal extraction process of antioxidant enzymes was determined to be 6.0 for pH,20 ℃ for processing temperature and 20 min for processing time.Antioxidant enzyme activities were in sequence of two stage cutting seedlings ＞ common cutting seedling ＞ tissue culture seedling.CONCLUSION Considering the contents of effective components and antioxidant enzyme activities,two stage cutting method is the best among 3 breeding modes.

Relationship between receptor tyrosine kinase Axl and apoptosis relative proteins in osteosarcoma / 临床与实验病理学杂志

Purpose To identify the role of tyrosine kinase receptor Axl for anti-apoptosis which was induced by cisplatin (DDP) and methotrexate (MTX) chemotherapy and to analyze the relationship between P-Axl and apoptosis-related proteins in osteosarcoma.Methods Osteosarcoma cell lines MG63,143B and U2OS were used in apoptosis assays,Axl siRNA transfection,cytotoxicity assays,cell cycle analysis,etc.A total of 41 cases of osteosarcom patients were included for immunohistochemistry of EnVision two-step staining and clinico-pathological relative analysis.TUNEL assay was performed in ten cases for apoptosis detection.Results Among the osteosarcoma cell lines,Gas6/Axl could obviously protect tumor cells from apoptosis induced by DDP and MTX (P ＜ 0.05).Axl siRNA transfection enhanced cell apoptosis,whereas Gas6 prone to function upon previous knockdown by Axl siRNA.Among the 41 cases,the positive rate of P-Axl,BCL-2,and Bax was 85.4％,70.7％,and 36.6％,respectively.In contrast,the positive rate of them was 22.2％,11.1％,and 11.1％ in osteofibrous dysplasia,respectively.The expression levels of these apoptosisrelated factors were significantly higher in osteosarcoma than in osteofibrous dysplasia (P ＜ 0.05).Through clinico-pathological analysis,there were significant relationships between the survival status and BCL-2 or Bax expression (P ＜ 0.05).Pearson correlation analysis demonstrated that BCL-2 was positively correlated to P-Axl with statistical significance (r =0.842,P ＜ 0.0001).By Cox univariate analysis,BCL-2 or Bax was correlated with the patients' prognosis.TUNEL assay also demonstrated that P-Axl high expression inhibited apoptosis in osteosarcoma tissues.Conclusion Gas6/Axl protects osteosarcoma cells from the apoptosis induced by DDP and MTX chemotherapy and inhibits apoptosis in osteosarcoma tissue,possibly through the regulation of apoptosis-related protein BCL-2.

Engineering a disulfide bond in the lid hinge region of Rhizopus chinensis lipase: increased thermostability and altered acyl chain length specificity.

The key to enzyme function is the maintenance of an appropriate balance between molecular stability and structural flexibility. The lid domain which is very important for "interfacial activation" is the most flexible part in the lipase structure. In this work, rational design was applied to explore the relationship between lid rigidity and lipase activity by introducing a disulfide bond in the hinge region of the lid, in the hope of improving the thermostability of R. chinensis lipase through stabilization of the lid domain without interfering with its catalytic performance. A disulfide bridge between F95C and F214C was introduced into the lipase from R. chinensis in the hinge region of the lid according to the prediction of the "Disulfide by Design" algorithm. The disulfide variant showed substantially improved thermostability with an eleven-fold increase in the t(1/2) value at 60°C and a 7°C increase of T(m) compared with the parent enzyme, probably contributed by the stabilization of the geometric structure of the lid region. The additional disulfide bond did not interfere with the catalytic rate (k(cat)) and the catalytic efficiency towards the short-chain fatty acid substrate, however, the catalytic efficiency of the disulfide variant towards pNPP decreased by 1.5-fold probably due to the block of the hydrophobic substrate channel by the disulfide bond. Furthermore, in the synthesis of fatty acid methyl esters, the maximum conversion rate by RCLCYS reached 95% which was 9% higher than that by RCL. This is the first report on improving the thermostability of the lipase from R. chinensis by introduction of a disulfide bond in the lid hinge region without compromising the catalytic rate.

Surgical treatment for cancer of the pancreatic head / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore and evaluate the therapeutic efficacy of surgical treatment for cancer of the pancreatic head.</p><p><b>METHODS</b>The clinical data of 96 patients with cancer of the pancreatic head admitted in our hospital from January 2002 to December 2009 were retrospectively analyzed. pancreatoduodenectomy was performed in 48 cases, extended pancreatoduodenectomy in 30 cases, and Roux-Y cholangiojejunostomy in 18 cases.</p><p><b>RESULTS</b>The 1, 2 and 3-year survival rates were 59.2%, 41.8% and 13.2%, respectively, in the patients treated with pancreatoduodenectomy, and 73.2%, 58.2% and 24.1%, respectively, in the patients treated with extended pancreatoduodenectomy. The 1, 2 and 3-year survival rates were 36.8%, 15.8% and 5.3%, respectively, in the patients with unresectable tumor who received radiotherapy and (or) chemotherapy in Roux-Y cholangiojejunostomy. The postoperative morbidity was 29.2%, 30.0% and 27.8% in the patients treated with pancreatoduodenectomy, extended pancreatoduodenectomy and Roux-Y cholangiojejunostomy, respectively.</p><p><b>CONCLUSIONS</b>Pancreatoduodenectomy is the most effective treatment. Extended pancreatoduodenectomy can improve the surgical resection rate, reduce the recurrence rate and improve the survival rate. Internal drainage is an important palliative measure.</p>

C-reactive protein level and the correlation between lung function and CRP levels in patients with chronic obstructive pulmonary diseases / 中南大学学报(医学版)

OBJECTIVE@#To investigate the changes of C-reactive protein (CRP) level in patients with chronic obstructive pulmonary diseases (COPD) and to analyze the correlation between the lung function and CRP levels.@*METHODS@#Sputum and serum specimens were obtained from 30 COPD patients diagnosed according to the national criteria. The CRP level was measured by enzyme-linked immunosorbent assays. The lung function indexes such as the forced expiratory volume in one second (FEV1) and the forced expiratory volume in one second/forced vital capacity (FEV1/FVC) were also determined.@*RESULTS@#The sputum CRP level in COPD patients was 30-50 times higher than that of the health controls. The sputum CRP level in COPD patients is significantly higher than that of serum in these patients (P < 0.05). The correlation between changes of serum and sputum CRP levels and sputum CRP levels was positive (r = 0.625, P < 0.05). The correlations between changes of serum CRP levels and FEV1 were negative (r = -0.610, -0.725, respectively, P < 0.001). And the correlations between changes of serum and sputum CRP levels and FEV1/FVC were negative (r = -0.639, -0.600, respectively, P < 0.05).@*CONCLUSION@#The CRP may be secreted from the local respiratory tract. The damage of lung function in COPD patients is associated with the increase of CRP level.