ATP-binding cassette transporter A1 (ABCA1) and scavenger receptor class B type I (SR-BI/CLA-1) are the key proteins in reverse cholesterol transport (RCT). The high expression of ABCA1 and SR-BI/CLA-1 can decrease the danger of atherosclerosis. The purpose of the study is to find ABCA1 and CLA-1up-regulators for treating atherosclerosis by using cell-based high throughput screening models. Among 20 000 compounds screened, E0869 [1-(3, 4-dimethylphenyl)-1-oxopropan-2-yll4-((methylsulfonyl)methyl)benzoate] was found as the positive hit. The up-regulated activities of E0869 in ABCAl1-LUC and bCA-l1-LUC HepG2 cell were 160% and 175%, respectively. The EC50 values of E0869 in ABCAl1-LUC and CLA-l1-LUC HepG2 cell were 3.79 and 1.42 pμol- x ,(-1) respectively. E0869 could upregulate the mRNA and protein levels of ABCA1, SR-BI/CLA-1 and ABCGJ1genes in HepG2 and RAW264.7 cells by Real-Time Quantitative PCR and Western blotting analysis, but could not influence the expression of FAS, SREBP-l1 and CD36. Foam cell assay showed that E0869 could inhibit lipids accumulation in mouse peritoneal macrophages RAW264.7. Cholesterol efflux assay showed that E0869 could induce HDL-mediated cholesterol efflux in mouse peritoneal macrophages RAW264.7. In conclusion, E0869 could up-regulate ABCA1 and CLA-1 activity, and had good anti-atherosclerotic activity in vitro.
ATP Binding Cassette Transporter 1 ; metabolism ; Animals ; Atherosclerosis ; drug therapy ; Biological Transport ; Cholesterol ; Hep G2 Cells ; High-Throughput Screening Assays ; Humans ; Macrophages, Peritoneal ; drug effects ; Mice ; RNA, Messenger ; Scavenger Receptors, Class B ; metabolism ; Up-Regulation

ObjectiveTo establish the time-resolved fluoroimmunoassay (TRFIA) method for the detection of serum galectin-3 and investigate the clinical value of serum galectin-3 for the diagnosis of pancreas cancer.MethodsMonoclonal anti-human galectin - 3 antibody and biotinylated polyclonal antibody were used to establish the sandwich TRFIA for detection of serum galectin-3.The optimal experimental condition was studied.Serum levels of galectin-3,CEA and CA19-9 in the patients with pancreatic cancer,benign pancreatic mass,pancreatitis,and healthy controls were measured.The diagnostic value of serum galectin-3,CEA and CA19-9 for pancreas cancer was studied.ResultsThe linearity of the TRFIA for detection of serum galectin3 tanged between 0 to 100 μg/L.The within-run CV and between-run CV were ≤6.45％ and ≤8.68％,respectively,and the average recovery was 106.6％.The level of serum galectin-3 was 4.93 ( 0.85 ～ 23.80) μg/L in pancreatic cancer group,which were significantly higher than those in benign pancreatic mass [2.83 ( 2.17 ～ 4.06) μg/L ],pancreatitis [ 2.62 (0.55 ～ 9.76 ) μg/L ],and healthy controls group [ 1.88 ( 0.59 ～ 3.94) μg/L] (P ＜0.05).By using 3.77 μg/L as the cut-off point,the smsitivity,specificity for the diagnosis of pancreatic cancer was 75.5％ and 90.9％.The levels of Gal 3 and CEA,CA19-9 was not correlated ( r =0.1321,P =0.3761 ; r =0.0920,P =0.5384).Combined determination of galactin-3 and CEA,CA19-9 levels could increase the diagnostic sensitivity to 92％.ConclusionsTRFIA method for the detection of galactin-3 is sensitive and stable.Galectin-3 could be a potentially novel serum tumor marker of pancreatic cancer.

Objective To study the value of serum galectin-3 detected by nanomagnetic beads sorting time resolved fluoroimmunoassay (TRFIA) tandem analysis in the diagnosis of pancreatic cancer.Methods The serum of 88 cases of pancreatic cancer,50 cases of acute pancreatitis,10 cases of pancreatic cysts,and 20 cases of healthy volunteers as control were collected.First,galectin-3 antibody coupled nanomagnetic-beads was used to sort the semm,then TRFIA was applied to detect the level of galectin 3,and the result was compared with that of routine TRFIA.ROC curve was constructed to determine the cutoff value and sensitivity for pancreatic cancer diagnosis.Results The method of nanomagnetic beads sorting TRFIA detected the level of galectin 3 between O.78 and 100 ng/ml in a linear manner,the intra-CV was ≤6.38％,and inter-CV was ≤7.22％,and average recovery rate was 105.3％.The serum level of galectin-3 in control group by nanomagnetic beads sorting TRFIA was 0.38 (0.02 ～ 3.06) ng/ml,which was significantly higher than that detected by routine TRFIA [0.18 (0.00 ～ 2.64) ng/ml,P =0.000).The serum levels of galectin-3 by nanomagnetic beads sorting TRFIA in pancreatic cancer,acute pancreatitis,pancreatic cysts and healthy volunteers were 4.85(0.00 ～42.67),0.69(0.00～ 13.62),0.70(0.00 ～ 14.54),0.38(0.02 ～ 3.06) ng/ml,and the level of galectin-3 in pancreatic cancer was significantly higher than that in acute pancreatitis,pancreatic cysts and healthy volunteers group (P ＜0.01),while the difference among the other three group was not significantly different.The AUC of galectin-3 for pancreatic cancer was 0.851 ± 0.040,95％ CI:0.772 ～ 0.929.While using 1.07 ng/ml as the cut-off value,the positive rates for the diagnosis of pancreatic cancer,acute pancreatitis,pancreatic cysts and healthy volunteers were 84.1％ (74/88),34.0％ (17/50),20.0％ (2/10),10.0％ (2/20),and the sensitivity for diagnosis of pancreatic cancer was significantly higher than those in other 3 groups (P ＜ 0.01).Conclusions Nanomagnetic beads sorting TRFIA tandem analysis method can enrich serum galectin-3,and increase the sensitivity of detection for pancreatic cancer and enhance the diagnostic value of galectin-3 for pancreatic cancer.

Objective To investigate the features and risk factors associated with lymph node metastasis in colorectal neuroendocrine neoplasm.Methods Clinical and pathological data of 63 patients with colorectal neuroendocrine neoplasm treated in Yinzhou People's Hospital between June 1997 to December 2012,were analyzed retrospectively.Comparisons of categorical data and univariate analysis of risk factors of lymph node metastasis were conducted by x2 test,multivariate analysis was conducted by Logistic regression analysis.Results Of the 63 patients the rate of regional lymph node metastasis was 30％ (19/63) with 58％ limited to para-intestinal lymph nodes in 11 cases,26％ limited to mesenteric lymph nodes in 5 cases,and 16％ limited to mesenteric root central lymph nodes in 3 cases.No metastasis exceeding central lymph nodes was observed.According to univariate analysis,tumor size,depth of invasion,ulceration in mucous membrane,invasion of lymphatic vessel and pathological grading suggested by WHO were related to regional lymph node metastasis (P ＜ 0.01).Multivariate regression analysis revealed that tumor size,invasion of lymphatic vessel and pathological grading were independent risk factors for lymph node metastasis in colorectal neuroendocrine neoplasm (P ＜ 0.05).Conclusions Colorectal neuroendocrine neoplasm with larger tumor size,invasion of lymphatic vessel or higher grade (G2,G3) has high risk of regional lymph node metastasis.

Objective To study the effect of mixed purified autogenic and allogeneic hematopoietic stem cell transplantation for the treatment of systemic lupus erythematosus. Methods Thirty-six MRL/lpr mice were randomly divided into the control group, the study group,the mixed group ( the ratio of autogenic to hematopoietic stem cells, mixed in different proportions were infused intravenously after 60Co irradiation. The study group were treated with daily intraperitoneal infusion of dexamethasone 1 mg·kg-1·d-1, while the control group were treated with intraperitoneal infusion of equivalent volume of saline daily. The changes of serum creatinine level, the urine protein excretion of the mice and blood WBC count were compared. Repeat measures ANOVA was used for data analysis. ELISA was used for anti-nuclear antibody detection Light microscopy, electronic micros-copy, immunofluorescence were applied to detect the pathological changes in renal tissue. Results Serum creatinine and urine protein excretion levels increased with time in the ontrol group, while those of the transplant group and the study group decreased. The reduction in mixed transplantation group and the study group was more evident compared with that of the allogeneic group. The difference was statistically significant (P＜0.05), but there was no significant difference between the mixed transplantation groups and the study group (P＞0.05). The histopathologic damage was most serious in the control group as pathological injury score of most mice were in grade 3 or 4. The majority of the histopathologic damage of the allogeneic group was in grade 2. Most f pathological damage of the study drug group and the mixed transplantation group were grade 1 or 2. Conclusion Mixed hematoopoietic stem cell transplantation for the treatment of murine systemic lupus erythematosus can effectively correct heavy proteinuria in murine systemic lupus erythematosus so improve the renal damage. It is a safe and effectively way to treat murine systemic lupus erythematosus.

OBJECTIVE:To observe clinical effects and safety of small dose of octreotide for preventing hyperamylasemia and acute pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).METHODS:One hundred and twenty ERCP pa tients were selected from our hospital during Oct.2014-Jan.2015 and then divided in to observation group and control group in accordance with random number table,with 60 cases in each group.Both groups were given Diazepam tablet 10 mg+Meperidine hydrochloride tablet 100 mg+Phenobarbital scopolamine tablet 2 tablets 0.5 h before surgery for sedation and analgesia,and routine acid suppression and anti-infective therapy.Observation group was additionally given Octreotide acetate injection 0.1 mg hypodermically and then given Octreotide acetate injection 0.1 mg immediately after surgery,8 h after surgery.The levels of serum amylase and blood glucose were observed in 2 groups,and the occurrence of postoperative complication and ADR were recorded.RESULTS:Before operation,there was no statistical significance in the levels of serum amylase and blood glucose between 2 groups (P＞ 0.05).After operation,the level of serum amylase in control group was significantly higher than in observation group,with statistical significance (P＜0.05).There was no statistical significance in blood glucose level between 2 groups after operation (P＞0.05).The incidence of hyperamylasemia and ADR in observation group was significantly lower than in control group,with statistical significance (P＜0.05),and there was no statistical significance in the incidence of acute pancreatitis between 2 groups after operation (P＞0.05).CONCLUSIONS:Small dose of octreotide can effectively reduce the level of serum amylase and the incidence of hyperamylasemia after ERCP with good safety.

In the previous study, a high-throughput screening method was established to find the antagonists of CD36. In the present study, a new compound named IMB-1680 was found using this method. The anti-atherosclerotic activities of IMB-1680 were then evaluated. Dose-dependent activities of IMB-1680 were detected by using Sf9 [hCD36] and CHO [hCD36] models. Fluorescence microscopic photography and flow cytometry were used to analyze uptake of mLDL. Foam cell test with RAW264.7 macrophages was used to examine lipid accumulation. The results showed that IMB-1680 inhibited CD36 activity with IC50 of 2.80 and 8.79 micromol x L(-1) in Sf9[hCD36] and CHO [hCD36] cells, respectively. Fluorescence microscopic photography and flow cytometry revealed that IMB-1680 could significantly reduce DiI-AcLDL uptake. Meanwhile, IMB-1680 also could reduce lipids accumulation in RAW264.7 macrophages. In all, the data indicated that IMB-1680 might be a potent effective anti-atherosclerotic leading compound.
Animals ; CD36 Antigens ; antagonists & inhibitors ; genetics ; metabolism ; CHO Cells ; Cells, Cultured ; Cricetulus ; Dose-Response Relationship, Drug ; Foam Cells ; cytology ; High-Throughput Screening Assays ; Humans ; Lipoproteins, LDL ; metabolism ; Macrophages ; cytology ; metabolism ; Mice ; Molecular Structure ; Plasmids ; Receptors, Scavenger ; antagonists & inhibitors ; Sf9 Cells ; Spodoptera ; Transfection

OBJECTIVETo investigate the impact of 1, 25-(OH)2D3 on left ventricular hypertrophy (LVH) in type 2 diabetic rats.

METHODSType 2 diabetic mellitus (DM) model rats were established by intraperitoneally injecting with 30 mg/kg streptozotocin. After 8 weeks, 19 male rats were identified as diabetic with left ventricular hypertrophy (LVH) by ultrasound examination, and randomly assigned into three groups: untreated (DM-LVH, n=7), treated with insulin (DM-LVH+INS, n=6), and treated with 1, 25-(OH)2D3 (DM-LVH+VD, n=6). Healthy male rats were used as the controls group (n=6). The fasting blood glucose and the insulin level were determined weekly. The left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor level were determined by 4 weeks later.

RESULTSIn the DM-LVH model group, the insulin level was significantly decreased compared with the non-diabetic control group (P<0.05), whereas the blood glucose, left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor expression were significantly increased (all P<0.05). In the DM-LVH+INS and DM-LVH+VD groups, the insulin levels were significantly increased compared with the DM-LVH model group (P<0.05), whereas the other parameters were significantly decreased (all P<0.05).

CONCLUSION1, 25-(OH)2D3 could reverse LVH in diabetic rats and that the mechanism may involve stimulating insulin secretion and reducing blood glucose via direct up-regulation of 1, 25-(OH)2D3-receptor expression.

Animals ; Blood Glucose ; analysis ; Calcitriol ; therapeutic use ; Diabetes Mellitus, Experimental ; blood ; complications ; Diabetes Mellitus, Type 2 ; blood ; complications ; Hypertrophy, Left Ventricular ; prevention & control ; Insulin ; blood ; Male ; Rats ; Rats, Wistar ; Receptors, Calcitriol ; analysis ; Streptozocin

Objective To investigate the therapeutic effects and mechanism of anti-radiation pneumonia decoction(ARPD) on radiation induced lung fibrosis in rats.Methods One hundred and five male SD rats in a SPF grade were divided into Chinese medicine group,single radiation group and control group by random digits table method,with 35 in each group.After anesthetization,rats in Chinese medicine and single radiation groups were exposed to 6 MV X-rays at the dose of 15Gy.Rats in Chinese medicine group were treated with ARPD at the dosage of 10 ml·kg-1 ·d-1 once a day,but rats in single radiation group did not receive ARPD treatment.Rats in control group were treated with neither irradiation nor drugs.Five rats of each group were killed and the lung tissues and blood samples were collected at 15,30,60,75,90,105 and 140 d.The pathological changes of lung tissues were observed and the tissue protein and gene expressions of TGF-β1,PAI-1 and collagen type Ⅲ(C Ⅲ) were assayed by Western blot and RT-PCR.ELISA was used to detect serum TGF-β1 and plasma PAI-1.Tissue and serum HYP were determined by acid hydrolysis and alkaline hydrolysis methods respectively.Results Inflammation was found in the lung tissues of all the exposed rats.Obvious pathological lung fibrosis was found at 60 d,the inflammation and the fibrosis in treated group were slighter than those in single radiation group.In Chinese medicine group,the protein and gene expression levels of TGF-β1,PAI-1,C Ⅲ 30 d(Protein:t =2.49-3.74,t =2.63-4.57 and t =2.76-3.83;Gene:t =2.59-4.33,t =2.83-4.62 and t =2.83-3.96,P＜0.05),serum TGF-β1 and plasma PAI-1 15 dlater (t =2.85-6.27 and t =3.69-5.27,P＜0.05),and the levels of tissue and serum HYP60 dlater (t=3.65-4.40 and t =6.56-3.75,P＜0.05),all of them were lower than those in single radiation groups.There were significant positive correlations between tissue TGF-β1 and PAI-1 as well as C Ⅲ (Protein expression:r =0.604,0.759,P ＜0.05;Gene expression:r=0.519,0.816,P＜0.05).Conclusions ARPD may inhibit the pulmonary fibrosis by decreasing the levels of TGF-β1,PAI-1 and C Ⅲ.

· AIM: To summarize the clinical features of Chinese primary congenital glaucoma and to investigate the discrepancies of the clinical features between the Chinese and Indian patients.congenital glaucoma patients were reviewed. The clinical features were summarized as several quantifiable clinical parameters and the severity of the disease was evaluated. Both the quantified clinical features and severity were statistically compared with those of the Indian patients, which were cited from the previous published articles.included in the study. In Chinese patients, sex ratio (male to female )was about 2:1, family history was presented in 3 patients (7.5%) and consanguinity was found in one patient (2.5%). The main symptoms and signs observed in Chinese patients spanned a wide spectrum of manifestations. The most frequent signs noted on the initial examination were enlarged eyeball (42.5%) and decreased visual acuity (35.0%). Compared with Indian patients, Chinese patients had a later onset, a delayed diagnosis, more severe corneal changes and more severe optic nerve damages (P＜0.01). The combined tabeculectomy and trabeculotomy operation was preferred by both Chinese and Indian doctors whereas a higher proportion of Indian patients received the combined operation (P＜0.01). The proportions of the severity grade were different between Chinese and Indian patients. Most Chinese patients were in the severe grade while most Indian patients were in the very severe grade (P＜0.01).patients were sporadic and non-consanguineous.Compared with Indian patients, Chinese patients had a relative later onset, a delayed diagnosis and treatment.More attempts are needed in Chinese PCG prevention and treatment.