Objective To evaluate the relationship between glucose-regulated protein 78 (GRP78) and diabetes mellitus-induced influence on myocardial protection provided by remifentanil postconditioning in vitro.Methods H9c2 cells were cultured in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum.The cells were seeded in 96-well (100 μl/well) or 6-well (2 ml/well) plates at the density of l05 cells/ml.The cells were then randomly divided into 6 groups (n =24 each) using a random number table:normoglycemic control group (group NC),normoglycemic hypoxia/reoxygenation (H/R) group (group NHR),normoglycemic remifentanil postconditioning group (group NRP),hyperglycemic control group (group HC),hyperglycemic H/R group (group HHR),and hyperglycemic remifentanil postconditioning group (group HRP).In NC,NHR and NRP groups,the cells were cultured in normoglyccmic culture medium (5.5 mmol/L) for 48 h.In HC,HHR and HRP groups,the cells were incubated in hyperglycemic culture medium (25.0 mmol/L) for 48 h.In NHR,NRP,HHR and HRP groups,after changing the culture medium for Tyrode solution,the cells were exposed to 95％ N2-5％ CO2 in an incubator at 37 ℃ for 5 h.Subsequently,in NHR and HHR groups,the culture medium was changed to DMEM/F12 culture medium supplemented with 10％ fetal bovine serum and glucose at the corresponding concentration,and the cells were incubated for 1 h;in NRP and HRP groups,the cells were incubated for 1 h in the DMEM culture medium containing remifentanil at the final concentration of 1 μmol/L.At 1 h of reoxygenation,the cells of 9 wells in each group were selected to measure the cell viability by CCK8 assay,the cells of 12 wells in each group were selected to determine the activity of lactic dehydrogenase (LDH) released in the supernatant using colorimetric method,and the cells of 3 wells in each group were selected to detect the expression of GRP78 by Western blot.Results Compared with group NC,the cell viability was significantly decreased,the LDH activity was increased,and the expression of GRP78 was up-regulated in group NHR.Compared with group NHR,the cell viability was significantly increased,the LDH activity was decreased,and the expression of GRP78 was down-regulated in group NRP,and the cell viability was significantly decreased,the LDH activity was increased,and the expression of GRP78 was up-regulated in group HRP.Compared with group HC,the cell viability was significantly decreased,the LDH activity was increased,and the expression of GRP78 was up-regulated in group HHR.There was no significant change in the parameters mentioned above between group HRP and group HHR.Compared with group NRP,the cell viability was significantly decreased,the LDH activity was increased,and the expression of GRP78 was up-regulated in group HRP.Conclusion Up-regulation of GRP78 expression may be involved in the mechanism by which diabetes mellitus negates myocardial protection induced by remifentanil postconditioning in vitro.

Gastric Lavage ; methods ; Humans ; Mannitol ; administration & dosage ; Norepinephrine ; administration & dosage ; Organophosphate Poisoning ; Pesticides ; poisoning ; Poisoning ; therapy

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers ; urine ; Diabetic Nephropathies ; complications ; drug therapy ; urine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Proteinuria ; complications

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies ; complications ; diagnosis ; urine ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteinuria ; complications ; urine ; Qi ; Regression Analysis ; Yin-Yang

OBJECTIVETo investigate the effects of methylprednisolone pretreatment on pulmonary lung permeability index and the content of the pulmonary surfactant dipalmitoylphosphatidylcholine (DPPC) in a rabbit model of reexpansion pulmonary edema.

METHODSTwenty-one male New Zealand white rabbits were randomly divided into control group, reexpansion, and reexpansion+methylprednisolone pretreatment groups. The rabbit model of reexpansion pulmonary edema was established using Sakaos method. A bolus dosage of methylprednisolone (3 mg/kg) in reexpansion+methylprednisolone group group or 2.0 ml/kg normal saline in the other two groups was administered intravenously 20 min before reexpansion pulmonary edema. Bronchoalveolar lavage fluid (BALF) and arterial blood samples were collected for measurement of the total protein (TP) and DPPC contents 4 h after reexpansion, and the pulmonary permeability index was calculated.

RESULTSThe pulmonary permeability index in methylprednisolone pretreatment group was significantly lower than that in the reexpansion group (0.007∓0.002 vs 0.177∓0.004, P<0.05). Methylprednisolone pretreatment significantly increased DPPC concentration in the BALF as compared with saline treatment in the reexpansion group (61.815∓28.307 vs 101.955∓24.544 µg/ml, P<0.05).

CONCLUSIONMethylprednisolone pretreatment can increase pulmonary surfactant content and improve pulmonary permeability in the rabbit model of reexpansion pulmonary edema.

1,2-Dipalmitoylphosphatidylcholine ; analysis ; Animals ; Bronchoalveolar Lavage Fluid ; Capillary Permeability ; drug effects ; Male ; Methylprednisolone ; pharmacology ; Permeability ; Pulmonary Edema ; metabolism ; physiopathology ; Pulmonary Surfactants ; metabolism ; Rabbits

OBJECTIVECT120B gene is a splicing variant of CT120A, which deletes 96 nucleotides and leads to an in-frame loss of 32 amino acids between the codon 136 and 167 as compared with CT120A. This study was undertaken to assess the effects of CT120B expression on lung cancer cell growth and to explore the gene expression profiles.

METHODSCT120B cDNA was transfected into the human lung adenocarcinoma SPC-A-1 cells, and stable cell lines overexpressing CT120B were established. CCK-8 assay and tumorigenecity in a xenograft model were performed to analyze cell proliferation in vitro and in vivo. The differential gene expression induced by overexpressed CT120B was investigated using Atlas cDNA expression array. Flow cytometry was performed to analyze cell cycle and cell apoptosis.

RESULTSOverexpression of CT120B in SPC-A-1 cells resulted in a reduced cell growth rate in vitro, and decrease of the tumorigenicity in nude mice. A total of 38 genes were identified as differential expressions with more than a 2.0-fold change by Atlas cDNA expression array analysis, including downregulated cyclin E1, cdk 2, c-kit, CXCR4 and upregulated caspase 8 gene. Overexpression of CT120B also induced G1 phase arrest, but had no effect on cell apoptosis.

CONCLUSIONThe G1 cell cycle arrest, but not apoptosis, underlay the growth inhibitory activities of CT120B. The down-regulation of c-kit and CXCR4 expression might also contribute to the suppressive effects on cell growth of CT120B.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; G1 Phase ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Proteins ; genetics ; metabolism ; Neoplasm Transplantation ; Oligonucleotide Array Sequence Analysis ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptors, CXCR4 ; metabolism ; Transfection

Objective CD4 +IL-17 +cells are a group of newly discovered effector CD4 +T cells, which may play a key role in the pathogenesis of cancer.This study aims to investigate the expres-sion of CD4 +IL-17 +cells in pancreatic cancer and its correlation with the clinicopathological characteristics and prognosis of the dis-ease as well as the clinical significance of the cells in the microenvironment of pancreatic cancer. Methods We collected tumor tis-sue and tumor-adjacent normal tissue samples from 51 pancreatic cancer patients.We determined the expressions of CD34 and vascular endothelial growth factor ( VEGF) and measured the proportion of IL-17 +cells in the cancer tissue using immunohistochemistry and the fluorescence activated cell sorter, respectively, followed by analysis of their correlation with tumor angiogenesis, clinicopathological pa-rameters, and survival time of the patients. Results The percentage of CD4 +IL-17 +cells in tumor tissue was positively correlated with microvessel density (r =0.534, P<0.05) and the expression of VEGF in the tumor tissue (r=0.356, P<0.05).IL-17 +cells were expressed more highly in the tumorous than in the tumor-adjacent normal tissue (P<0.05), and the expression level was correla-ted with the stage of tumor, node, and metastasis (TNM) and lymph node metastasis (P<0.05), but not with the patients′gender or age, tumor size, tumor location, histological grade, or local invasion (P>0.05).Fifty (98.0%) of the patients were successfully followed up for 2-67 months, which revealed a median survival time of 16.6 ±4.8 months, significantly longer in those with a higher expression of intratumoral IL-17 +cells (P<0.05).Univariate analysis showed an association of the survival rate with the tumor size, TNM stage, lymph node metastasis, and level of intratumoral IL-17 +cells, while multivariate analysis showed the TNM stage to be an independent prognostic factor for the survival of the pancreatic cancer patients. Conclusion The expression of CD4 +IL-17 +cells in the tumor tissue is positively correlated with tumor angiogenesis, while that of IL-17 +cells with the clinicopathological parameters and survival time of the patients and therefore may serve as an important immune indicator for the prognosis of pancreatic cancer.

OBJECTIVETo determine the enhancement effects of caffeine on chemotherapy of transplanted osteosarcoma in Fischer 344/N rats.

METHODSOsteosarcoma-bearing Fischer 344/N rats were treated with cisplatin 2.5 mg/kg (Group DDP), caffeine 90 mg/kg x 2 d (Group caffeine), and cisplatin 2.5 mg/kg plus caffeine 90 mg/kg x 2 d (Group DDP+caffeine), and the control group was treated with normal saline in the same volume. All drugs were given by intra-peritoneum injection with micro-pump, in the rate of 0.5 ml/h. The tumor volume was measured and evaluated. The tumors were stained in TUNEL, and PCNA was detected with immunohistochemistry. The tumor growth inhibition rate, PCNA index and apoptosis index were calculated, and the survival time were recorded.

RESULTSThe tumor inhibition rate was -0.5219 +/-0.1429 in control group, 0.0362 +/-0.0957 in Group DDP, -0.4193 +/-0.1345 in Group caffeine, and 0.3646 +/-0.1313 in Group DDP+caffeine (P <0.01). PCNA index was 0.4587 +/-0.1312 in control group, 0.1847+/-0.0535 in Group DDP, 0.4381 +/-0.0706 in Group caffeine, and 0.0314 +/-0.0231 in Group DDP+caffeine (P <0.01). Apoptosis index was 0.0008 +/-0.0005 in control group, 0.0077 +/-0.0060 in Group DDP, 0.0011 +/-0.0003 in Group caffeine, and 0.0295 +/-0.0069 in Group DDP+caffeine (P <0.01). And the survival time was (33.63 +/-4.63)d in control group, (52.13 +/-11.74)d in Group DDP, (35.63 +/-5.15)d in Group caffeine, and (55.13 +/-16.23)d in Group DDP+caffeine (P <0.01).

CONCLUSIONCaffeine could enhance the anti-tumor effect of cisplatin in rat osteosarcoma.

Animals ; Antineoplastic Agents ; therapeutic use ; Bone Neoplasms ; drug therapy ; pathology ; Caffeine ; therapeutic use ; Cisplatin ; therapeutic use ; Drug Synergism ; Neoplasm Transplantation ; Osteosarcoma ; drug therapy ; pathology ; Rats ; Rats, Inbred F344

Diosgenin can inhibit the growth of A375 and K562 cell lines and induce their apoptosis with an effect on pro-apoptotic members of Bcl-2 family. To study the SAR of diosgenin derivatives, and to improve the anti-tumor activity of diosgenin, a series of novel diosgenin derivatives were designed and synthesized. Their anti-tumor activities in vitro were evaluated. The results revealed that most of the new derivatives had potent effects against K562, A375 and A549 (three tumor cell lines) in vitro, and had no or less effect against H293 and L02 (two normal cell lines). Particularly, some compounds (e.g. 1, 6-8) showed excellent activities on K562 with IC50 values ranging from 1.96 to 4.35 micromol x L(-1).
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Diosgenin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Drug Design ; Humans

Objective To investigate the levels and clinical significance of serum miR-150 in children with primary nephrotic syndrome(NS).Methods Serum samples were collected from 78 NS children and 79 age-and sex-matched control children in Nanjing General Hospital,Jiangsu Province Hospital of TCM and Nanjing Children Hospital from March 2010 to May 2014.Quantitive Real-time PCR(qRT-PCR)assays were used to determine the concentrations of serum miR-150 in NS and control children.The other lipid and renal function parameters including serum TP,ALB,GLO,TC,TG,Urea,Cr,Uric acid and Uric protein were also assessed.Statistical analyzes were used to evaluate the clinical value of serum miR-150 for NS as well as to assess the clinical association between the levels of serum miR-150 and other clinical parameters.Results The ser-um levels of miR-150 were significantly elevated in NS children[101.4(21.29~336.6)fmol/L(F=3.658,P<0.001)]as compared with controls[34.11(5.53~134.2)fmol/L].ROC curve analysis showed that the area under the receiver operat-ing characteristic curve(AUCROC)was 0.892(95% CI=0.843~0.940).Spearman rank correlations analyzes showed that the levels of serum miR-150 were significantly negatively associated with GLO(r=-0.231,P=0.042)and TG(r=-0.233,P=0.040)in NS children.Multiple linear regression analyses showed that serum miR-150 was independently associ-ated with serum ALB levels(β=0.241,P=0.034;adjusted r2=0.046)after adjustment of other related factors.Further-more,multivariate logistic regression analysis showed that serum miR-150 an independent risk factor for NS after adjusting other factors including age and gender[OR=16.07(95% CI=5.35~48.28),P<0.001].Conclusion The serum levels of miR-150 were markedly elevated in NS children and closely associated with with impaired kidney function as well as lipid pa-rameters,and may have the potential as a novel auxiliary diagnosis marker for assessing the development of NS.