Objective To investigate the relationship between inflammatory bowel disease (IBD) activity and choroidal thickness,and evaluate the utility of a choroidal thickness measurement in assessing IBD activity.Methods A total of 100 eyes of 50 patients of IBD with different disease activity,including 23 patients of ulcerative colitis,27 patients of Crohn's disease (CD).Ninety-six eyes of 48 healthy volunteers were recruited as control group.Choroidal thickness was measured using enhanced depth imaging(EDI)optical coherence tomography.Results Compared with the subfoveal choroidal thickness (294.37 ± 35.04) μm in healthy volunteers,the subfoveal choroidal thickness (349.28 ± 76.57) μm in UC patients with severe disease activity,the subfoveal choroidal thickness (326.71 ± 59.71) μm and (354.24 ± 66.34) μm,respectively,in CD patients with moderate and severe disease activity were found to be increased significantly (all P ＜ 0.05).Conclusion Choroidal thickness should be considered as a potential marker to assess the disease activity in patients with IBD,especially in patients with CD.

Objective To investigate the clinical value of bone metabolism biochemical marker N-MID,TP1NP and beta-CTx combined with whole body bone scintigraphy in early diagnosis of bone metastasis of tumor.Methods The concentration of the 3 markers were measured by the electrochemical luminescence analysis method in 30 cases of healthy control group and 210 cases of patients with malignant tumor,which were divided into non bone metastasis group(45 cases) and bone metastasis group(165 cases).The bone metastasis group were divided into 4 grades(0-grade Ⅲ) by Soloway classification according to whole body bone imaging.Results The levels of serum N-MID,TP1NP and beta-CTx in 165 malignant tumor patients with bone metastasis were significantly higher than in 45 malignant tumor patients with bone metastasis and in 30 healthy control group,the difference was statistically significant(P<0.05).With the increase of the number of metastatic lesions in the bone metastasis group,the serum levels of N-MID,TP1NP,and beta-CTx were increased gradually,and they were positively correlated with the progression of the disease.According to the analysis of ROC curve,the cut-off value,sensitivity and specificity in the diagnosis of tumor bone metastasis were 17.59 ng/mL,70.3％,88.9％ for serum N-MID,43.04 ng/mL,78.2％,95.6％ for TP1NP,and 0.48 ng/mL,73.9％,93.3％ for beta-CTx.Under the ROC curve(AUC) was 0.831 for serum N-MID,0.890 for TP1NP,and 0.869 for beta-CTx.The sensitivity and specificity of three bone metabolic markers in the diagnosis of bone metastasis of malignant tumor were significantly higher.Conclusion Bone metabolism biochemical markers:Serum N-MID,TP1NP and beta-CTx for diagnosis of bone metastasis of malignant tumor are sensitive,accurate and simple,which can significantly improve the efficiency of diagnosis of bone metastasis,and can be combined with whole-body bone scintigraphy in early diagnosis of bone metastasis with malignant tumor.

OBJECTIVETo evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin.

METHODSRats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups.

RESULTSConsistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point.

CONCLUSIONAmygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.

Amygdalin ; pharmacology ; Animals ; Biomarkers ; blood ; Bleomycin ; adverse effects ; Blood Proteins ; metabolism ; Male ; Pulmonary Fibrosis ; blood ; chemically induced ; Rats ; Rats, Wistar

OBJECTIVEGlucocorticoid (GC) has occupied a central role in the treatment of acute lymphoblastic leukemia due to its ability to induce apoptosis in neoplastic lymphoid cells. Glucocorticoid resistance is present among 20% initial acute lymphoblastic leukemia, even 80% refractory acute lymphoblastic leukemia. Glucocorticoid resistance has been an important determinant of clinical outcome. Glucocorticoid depends on glucocorticoid receptor (GR) to induce apoptosis. Glucocorticoid receptor, a number of nuclear hormone receptor superfamily, is mediated by many signal transduction systems. The mitogen-activated protein kinases (MAPK) superfamily of serine/threonine kinases has emerged as an important component of cellular signal transduction. Four MAP kinase families, ERK, p38 MAP kinase, JNK, and ERK5, have been well characterized. p38 MAPK usually plays a role in regulating apoptosis, cell cycle arrest and cytokines production, et al. In steroid resistance patients, IL-2 combined with IL-4 can decrease glucocorticoid receptor ligand-binding affinity via p38MAPK. In human alveolar epithelial A549 cells, dexamethasone could inhibit the activation of p38MAPK. It is unclear that the effect of p38MAPK on glucocorticoid receptor function induced by dexamethasone in CEM cells. This study aimed to investigate effect of p38 mitogen-activated protein kinase on glucocorticoid receptor function induced by dexamethasone in CEM cells.

METHODSCell viability was determined by trypan blue dye exclusion. Apoptosis was evaluated by morphology and flow cytometry. Glucocorticoid receptor protein and p-p38MAPK protein were analyzed by Western Blot.

RESULTSWhen treatment with SB203580 and dexamethasone for 24 h to 72 h, the survival percentage was increased from 62.3%, 35.5% and 11.6% to 82.8%, 54.7% and 48.1%, respectively (P < 0.01). Co-treatment with SB203580 and dexamethasone resulted in the decrease of apoptotic percentage from 26.2% to 7.1% for 36 h (P < 0.01). p38 MAPK activation was apparent at 15 min, peaked at 1 h after dexamethasone treatment, and was sustained for 6 h. The phosphorylation was still observed at 48 h. Treatment with dexamethasone at 5 micromol/L for 12, 24, 36 and 48 h resulted in increase of GR(alpha) protein to 117%, 121%, 122% and 125% respectively. Unbinding to dexamethasone, GR(alpha) is in the cytoplasm. Nuclear-to-cytoplasmic ratio of GR(alpha) is 0.27. Treatment with dexamethasone at the same concentration and time resulted in the nuclear-to-cytoplasmic ratio increase to 0.48, 0.59, 0.95, 2.16 and 4.08 respectively. Combined treatment with SB203580 and dexamethasone resulted in the nuclear-to-cytoplasmic ratio decrease from 4.08 to 0.43 for 48 h (P < 0.05). The total GR(alpha) protein was unaffected.

CONCLUSIONSExpression of GR(alpha) protein is upregulated and translocated into nucleus. p38MAPK enhances GR(alpha) protein translocation into nucleus.

Apoptosis ; drug effects ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Enzyme Activation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Glucocorticoids ; pharmacology ; Humans ; Imidazoles ; pharmacology ; Mitogen-Activated Protein Kinases ; metabolism ; Phosphorylation ; drug effects ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; Protein-Serine-Threonine Kinases ; Pyridines ; pharmacology ; Receptors, Glucocorticoid ; drug effects ; metabolism ; Signal Transduction ; drug effects ; p38 Mitogen-Activated Protein Kinases ; drug effects ; metabolism

OBJECTIVETo observe the effect of Zishen Huoxue Recipe (ZHR) on pathomorphology in coronary heart disease (CHD) rats with Shen deficiency blood stasis syndrome (SDBSS).

METHODSTotally 60 healthy Wistar rats were divided into the blank control group, the model group, high, middle, and low dose ZHR groups according to random digit table, 12 in each group. Myocardial ischemia SDBSS rat model was prepared by ligating the left anterior descending coronary artery and injecting hydrocortisone. ZHR physic liquor was administered to rats in high, middle, and low dose ZHR groups at the daily dose of 21.6, 10.8, 5.4 g/kg by gastrogavage for 7 successive days, equal volume of pure water was administered to rats in the blank control group and the model group by gastrogavage for 7 successive days. Rat heart was collected for pathomorphological observation under light microscope.

RESULTSIn the model group the heart muscle fiber was swollen and deformed with widened space, loose and dropsy tissues. Blood vessels in myocardial mesenchymal were dilated, infiltrated with more inflammatory cells. Myocardial cells were markedly swollen, degenerated, or necrotic, with caryolysis or disappearance of partial nuclear. A large amount of collagen fibrous tissue became hyperplasia. Endocardial blood vessels were swollen and degenerated with infiltration of few inflammatory cells. Epicardium tissue and structure were destroyed and got hyperplasia. Swollen, degenerated, or necrotic vessels could be seen, with infiltration of more inflammatory cells and collagen deposition. Pathomorphological injuries were alleviated in each ZHR group. The higher ZHR concentration, the milder the injury degree of myocardial tissue, the more limited range of damage.

CONCLUSIONZHR could attenuate pathomorphological injuries of myocardial ischemia rats with SDBSS and regulate myocardial function, thus improving myocardial ischemia in CHD rats with SDBSS.

Animals ; Coronary Artery Disease ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Medicine, Chinese Traditional ; Myocardial Ischemia ; Myocardium ; Rats ; Rats, Wistar

OBJECTIVETo compare thoracoscopic esophagectomy with traditional esophagectomy in radical mediastinal lymphadenectomy for esophageal cancer, and to explore the feasibility and safety of thoracoscopic mediastinal lymphadenectomy for esophagectomy.

METHODSClinical data associated with perioperation and mediastinal lymph nodes clearance of 304 patients undergoing radical operation of esophageal cancer via left neck-right chest-upper abdomen in our department from June 2009 to June 2011 were analyzed retrospectively. Among 304 cases, 199 received traditional open radical resection and 105 thoracoscopic esophagectomy. The intrathoracic mediastinal lymph node metastasis rate, extent of metastasis, time of operation, blood loss and complications between two groups were compared.

RESULTSAll the 304 cases completed their operations successfully. A total of 3724 mediastinal lymph nodes were removed, mean 12.3±7.0 per case, including 1065 in thoracoscopic group, mean 10.1±5.5 per case, and 2659 in open group, mean 13.3±7.5 per case, whose difference was significant. But further analysis according to the postoperative pathologic staging showed no significant difference of above lymph nodes removed between two groups. Mediastinal lymph node metastasis was found in 126 patients with a rate of 41.4%, which was 35.6% and 44.7% in thoracoscopic and open groups respectively without significant difference(P>0.05). The left laryngeal recurrent nerve lymph node metastasis rate in open group and thoracoscopic group was 16.1% and 6.7% respectively, and the difference was significant(P<0.05). Differences of lymph node metastasis rate in other regions were not significant between the two groups. There were 365 positive lymph nodes, and the lymph node metastasis degree was 9.8%. which was 8.2% and 10.5% in thoracoscopic group and open group respectively(P<0.05), besides metastasis degree of open group was much higher in right laryngeal recurrent nerve and subcarinal lymph node region. The overall complication rate was 36.8%, which was 28.6% in thoracoscopic group and 41.2% in open group respectively with significant difference(P<0.05). There were no significant differences in operative time and blood loss between the two groups(both P>0.05).

CONCLUSIONRadical mediastinal lymphadenectomy with thoracoscopic esophagectomy is technically safe and feasible for early to moderate stage esophageal cancer with similar lymph nodes removed and lower complication morbidity. In the early period of carrying out thoracoscopic radical mediastinal lymphadenectomy, laryngeal recurrent nerve and subcarinal lymph node region should be identified to prevent incidental injury.

Blood Loss, Surgical ; Esophageal Neoplasms ; pathology ; surgery ; Esophagectomy ; methods ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Mediastinum ; pathology ; Operative Time ; Retrospective Studies

OBJECTIVEThe precise mechanism of glucocorticoid-induced apoptosis has not yet been elucidated. Survivin, a member of the inhibitors of apoptosis protein family, correlates with inhibition of apoptosis, proliferation, angiogenesis and multiple drugs resistance. This study aimed to investigate the variation of the survivin gene expression in apoptosis induced by dexamethasone (Dex) in the human T-lineage acute lymphoblastic leukemia (ALL) cell line, CEM-WT cells.

METHODSThe logarithmically growing CEM cells cultured in vitro (cell density 2 x10(5)/mL) were exposed to 0.1, 0.5, 1, 5, and 10 microM Dex, then were collected 24, 48 and 72 hrs later. Untreated CEM cells were used as Controls. The cell viability was determined by trypan blue dye exclusion. Apoptosis was evaluated by morphology and flow cytometry. Survivin protein and gene were analyzed by Western Blot and RT-PCR.

RESULTSCEM cells growth was obviously inhibited by 0.1, 0.5, 1, 5, and 10 microM Dex from 48 hrs. The inhibition effect was dose- and time-dependent. CEM cells treated with Dex (> or = 5 microM) exhibited typical apoptotic features. The apoptosis increased after 5 microM Dex treatment in a time-dependent manner, with the apoptosis percentage increasing from 14.9% (12 hrs) to 46.2% (48 hrs). Compared with that of the Control group, the expression of survivin protein was down-regulated, with the expression rate of 54.6%, 45.5%, 15.8% and 9.7% respectively at 12, 24, 48 and 72 hrs after 5 microM Dex treatment. 5 microM Dex treatment also resulted in a decrease of survivin mRNA expression. The survivin mRNA expression was 76.4%, 67.3%, 55.0%, 49.9%, 38.3% and 18.3% of the Control respectively at 6, 12, 24, 48 and 72 hrs after Dex treatment.

CONCLUSIONSApoptosis induced by Dex in CEM cells is associated with downregulation of the survivin expression.

Apoptosis ; drug effects ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; Leukemia-Lymphoma, Adult T-Cell ; metabolism ; pathology ; Microtubule-Associated Proteins ; analysis ; genetics ; Neoplasm Proteins ; analysis ; genetics

To produce specific monoclonal antibody (McAb) against human thrombomodulin (hTM), the full-length hTM cDNA-expressing plasmid pThr402 was transfected into CHO cells by Lipofectamine 2000 reagent. The hTM-expressing CHO cells, which was confirmed by flow cytometry and Western blot, were obtained by G418 selection. Then the McAb against hTM was prepared with classic hybridoma technique. A cell line of CHO-TM5 with high level of hTM was used to immunize female Balb/c mice 3 times at an interval of 4 weeks. On the third day after the third immunization, mice were sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96-well plates for screening. Cellular enzyme-linked immunoabsorbent assay (CELISA) was applied twice. The first CELISA was done with polythene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but with CHO cell monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5(+)CHO(-) hybridoma cells. Balb/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded McAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. One line of hybridoma cells with high expression of specific McAb against hTM, NH-1, was obtained. The Ig subclass of the McAb was IgG1 and the titer of ascitic McAb was 1x10(-6). Flow cytometry, CELISA and Western blot assays demonstrated that McAb NH-1 could specifically recognize hTM expressed in CHO-TM5 cells and human umbilical vascular endothelial cells. Meanwhile, the tissue specificity of antigen recognized by McAb NH-1 was identified by immunohistochemical ABC staining. NH-1 can specifically recognize the natural hTM expressed mainly in vascular endothelial cells, which will potentially be useful for investigation of the functions and clinic values of hTM.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; CHO Cells ; Cricetulus ; Female ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Thrombomodulin ; immunology ; Transfection

OBJECTIVEDiagnosing neonatal asphyxia solely according to Apgar score may lead to misdiagnosis. The aim of this study was to explore new and more accurate diagnostic criteria for neonatal asphyxia.

METHODSTotally 10 376 live born neonates in our hospital were consecutively enrolled into the study. The following five items related to birth asphyxia, i.e., antepartum high-risk factors, Apgar scores, umbilical artery blood pH, organ injury, differential diagnosis on the causes of low Apgar score cases were examined and registered. The relationship among the first 4 items were analyzed. By differential diagnosis, the sensitivity and specificity of each index on diagnosing asphyxia and their complementary value on each other were investigated.

RESULTSThe items correlated well with each other (P < 0.01 or < 0.05) but were not entirely parallel and consistent; they could complement but could not substitute for each other. The sensitivity of antepartum high-risk factors, low Apgar scores, umbilical artery blood pH < 7.00 and organ injury was 100%, 100%, 44.44% and 100%, while the specificity was 17.99%, 98.90%, 96.05% and 96.62%, respectively. Of the 230 low Apgar score cases in this series only 50.9% coincided with asphyxia. For the 230 cases, when low Apgar score was combined with umbilical artery blood pH < 7.00, the sensitivity and specificity were 41% and 99.1% and when low Apgar score was combined with umbilical artery blood pH < 7.20, the sensitivity and specificity were 100% and 29.20%, respectively. After organ injury was added, the specificity was increased to 65.49%. When differential diagnosis was further added to exclude the other causes of low Apgar score cases, the misdiagnosis rate was minimized.

CONCLUSIONUp to now, no single accurate index for diagnosing neonatal asphyxia is available. In order to increase diagnostic bases and reduce misdiagnosis, the criteria of sole Apgar score should be replaced by multi-index diagnostic criteria. Based on the present study, a set of integrated diagnostic criteria for neonatal asphyxia is proposed: (1) prenatal high-risk factors, (2) low Apgar scores (respiratory depression must present), (3) umbilical artery blood pH < 7.00, if only pH < 7.20, the items (2) (4) (5) must be present, (4) hypoxic-ischemic organ injury (at least one organ dysfunction), (5) the other causes of low Apgar scores should be excluded. The last 4 indexes should all be met and the first one serves as reference. If multi-organ (three or more organs) dysfunction and (or) hypoxic-ischemic encephalopathy are present, severe asphyxia can be diagnosed.

Apgar Score ; Asphyxia Neonatorum ; blood ; diagnosis ; Diagnosis, Differential ; Diagnostic Errors ; prevention & control ; Humans ; Hydrogen-Ion Concentration ; Infant, Newborn ; Multiple Organ Failure ; Risk Factors ; Sensitivity and Specificity

The extraction of active ingredient group, i.e., prim-o-glucosylcimifugin, astragaloside, and 5-o-methylvisammiosode from Yupingfeng powder with one-pot method was studied in this work. A HPLC method was used to determine the content of each active constituent mentioned above in the extract. The influences of extraction temperature, time, volume percent of ethanol in water and its amount added on the content and yield of active ingredient group were investigated by orthogonal test. The experimental results showed that the optimized extraction conditions were as follows: 1g of Yupingfeng powder was one-pot extracted for 4 hours at 80 degrees C with 90% ethanol as solvent, and the yield and content of active ingredient group were 0.16%, 0.53% respectively. The active ingredient group in Yupingfeng powder could be effectively one-pot extracted under the conditions above.
Apiaceae ; chemistry ; Astragalus membranaceus ; chemistry ; Atractylodes ; chemistry ; Chemical Fractionation ; methods ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Monosaccharides ; analysis ; isolation & purification ; Plants, Medicinal ; chemistry ; Powders ; Saponins ; analysis ; isolation & purification ; Temperature ; Triterpenes ; analysis ; isolation & purification ; Water ; chemistry ; Xanthenes ; analysis ; isolation & purification