Objective To investigate the role of early enteral nutrition on TLR 4 signaling pathway in rats with acute necrotizing pancreatitis (ANP).Methods Sixty SD rats were randomly divided into three groups:sham operation group ( SO group ) , total parenteral nutrition group ( TPN ) , early enteral nutrition group ( EEN ) .One day after ANP model induction , the serum level of amylase was measured .Nutrient solution was given for five days , then the rats were sacrificed , and the blood , pancreas and colon tissue were collected.The serum levels of IL-6, TNF-αwere detected by ELISA .Pathologic changes of pancreas were observed by HE staining.The intestinal TLR4, NF-κB expression was measured by Western blot .Results Mortality rates of SO group, TPN group, EEN group were 0, 50%, 25%, respectively; the serum levels of amylase were (744 ±41), (3 278 ±219), (2 227 ±169) U/L, respectively;the serum levels of TNF-αwere (81.57 ±18.25), (465.72 ±42.47), (223.21 ±29.94)ng/L, respectively; the serum levels of IL-6 were (362.83 ±41.32), (932.46 ±57.21), (628.62 ±142.24) ng/L, respectively; the pancreatic pathologic scores were (0.91 ±0.15), (11.1 ±0.61), (6.9 ±0.62);the intestinal TLR4 expressions were 0.7506 ± 0.003, 1.3404 ±0.004, 0.9544 ±0.004;the intestinal NF-κB expressions were 1.33 ±0.50, 6.92 ±1.06,2.93 ±0.89.The values of TPN and EEN group were significantly higher than those of SO group (P<0.05). The values of EEN group were significantly lower than those of TPN group (P<0.05).Conclusions EEN can inhibit TLR4 and NF-κB signal pathway in gut , then reduce IL-6 and TNF-αexpression and relieve inflammatory reaction of ANP , finally decrease the mortality of ANP .

A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

AIM: To investigate the role of retinal pigment epithelium (RPE) in the growth of Müller cells using a co-culture system in vitro . METHODS: Müller cells were cocultured with RPE cells under both normoxic and hypoxic conditions in Transwell chamber culture system. Müller cell proliferation was evaluated by MTT assay. The number of cells which migrate through micropores and stay on the outer bottom side of insert systems were observed and counted. RESULTS: The activities of proliferation and migration of Müller cells when cocultured with RPE cells were significantly higher than those of the Müller cells when cultured alone at all time points under both normoxic and hypoxic conditions. However, for both the coculture and control groups, there is no significant difference between the measurements at 3 and 6 hours. CONCLUSION: Evidence suggests that RPE, when co-cultured with Müller cells, can stimulate migration and proliferation of Müller cells under both hypoxic and normoxic conditions in a time-dependent manner; how-ever, there is no evidence to support the synergetic interaction of RPE and Müller cells co-cultured under hypoxic conditions.

OBJECTIVETo observe the inhibiting effect of acupuncture on blood lipid, myocardial hypertrophy and fibrosis in mice with hyperlipemia, and explore its possible action mechanism.

METHODSTen inbred mice (C57) were applied. Forty ApoE(-/-) mice who removed gene of apolipoprotein E were randomly divided into a control group, a non-acupoint group, an acupoint group and a medication group. The points 0. 5 cm and 1 cm next to the end of mice tail were respectively punctured in the non-acupoint group; "Neiguan" (PC 6) and "Fenglong" (ST 40) were punctured in the acupoint group; intragastric administration of simvastatin was applied in the medication group. After 8 weeks of treatment, the changes of total cholesterol (TC) and ratio of heart to body mass in each group were measured; changes of cardiac muscle fiber and ventricular wall thickness were observed; enzyme linked immunosorbent assay (ELISA) was used to test the level of angiotensin II (Ang I ) in plasma, and western blotting method was used to test protein content of angiotensin II type 1 receptor (AT1R) and endothelin-1 type A receptor (ETAR) in the heart.

RESULTSAfter 8 weeks of intervention, compared with the control group, rising range of blood lipid was obviously decreased (P<0.01) in the acupoint group and medication group, ratio of P<0.01), myocardial heart to body mass was decreased (P<0.05), thickness of ventricular wall was reduced (P fibrosis was relieved, levels of Ang II and ET-1 in plasma were decreased (P<0. 05), content of NO was increased (P<0. 05), and protein content of AT1R and ETAR was decreased in the heart (P<0. 05).

CONCLUSION40) could inhibit the rising of blood lipid in ApoE(-/-) mice, lower the levels of Ang II and ET-1 in peripheral blood, increase the content of NO and inhibit the expression of AT1R and ETAR in heart tissue, which could relieve myocardial hypertrophy and fibrosis to play a protective role on heart.

Acupuncture Points ; Acupuncture Therapy ; Angiotensin II ; metabolism ; Animals ; Blood Pressure ; Disease Models, Animal ; Heart ; physiopathology ; Heart Diseases ; etiology ; metabolism ; physiopathology ; prevention & control ; Humans ; Hyperlipidemias ; complications ; physiopathology ; therapy ; Male ; Mice ; Mice, Inbred C57BL ; Myocardium ; metabolism

Objective To study the effects of insulin therapy on alterations of learning and memory functions and somatostatin (SS) and vasoactive intestinal peptide (VIP) levels in frontal cortex and hippocampus of diabetic rats. Methods The diabetic rat models were produced by injection with 1% streptozotocin, followed by insulin administration. The water maze test was used to verify the changes of learning and memory functions of the rats and radioimmunoassary was carried out to observe the alterations of SS and VIP levels in the cerebral cotex and hippocampus. Results The swimming time of untreated diabetic rats in water maze was prolonged significantly and the successful frequencies decreased significantly in comparison to those of controls. The contents of SS reduced significantly both in the frontal cortex and hippocampus of untreated diabetic rats, while VIP were not changed. The changes in learning and memory functions and SS levels in insulin-treated diabetics were not significantly different from those of controls. Conclusion The cognitive dysfunctions induced by diabetes may be correlated with the decrease of SS and alleviated with insulin administration.

·AIM:To observe the clinical effect of bilateral oblique muscle weakening on the A - V patterns strabismus correction, and dose - response relationship between bilateral oblique muscle weakening and A-V syndrome degree.·METHODS: Totally 70 patients ( 70 eyes ) with A-V syndrome treated in our hospital from January 2012 to January 2016 were enrolled in this study, including 28 eyes of A sign and 42 eyes of V sign. All patients underwent bilateral oblique muscle reduction combined with horizontal rectus correction. The changes of the upper and lower strabismus were observed before and after operation, and the patients were followed up for 18mo. The successful rate of A-V sign and strabismus correction were observed and counted. In addition, the quantitative relationship between the degree of A-V and the degree of correction and the amount of residual after operation was compared and analyzed.·RESULTS: After operation, the oblique eye position angle difference of A - esotropia, A - exotropia, V -esotropia and V-exotropia were respectively -1. 98△ ± 3. 92△, -2. 25△± 4. 88△, 2. 96△± 5. 29△, 1. 91△± 4. 67△, the differences were significantly compared with preoperatively ( P < 0. 05 ). The reduce amount after operation were 25. 10△± 9. 79△, 24. 29△± 8. 63△, 23. 81△± 9. 24△, 22. 79△±8. 95△. After 18mo follow-up, the success rate of A sign correction was 96% ( 27/28 ) , and the successful rate of V sign correction was 93% ( 39/42 ) . A-V sign, the more the oblique angle before operation, the more the correction amount during the weakening of bilateral oblique muscle, and the postoperative oblique residual increased accordingly.·CONCLUSION: Bilateral oblique muscle weakening can effectively treat strabismus with A - V sign, and can significantly reduce the upper and lower ocular gradient. The upper and lower oblique angle of the A- V sign increase, and the corrected angle and residual volume increase with the quantitative relationship.

OBJECTIVETo investigate the significance of Th17/Treg imbalance in the development and treatment of primary immune thrombocytopenia (ITP) in children.

METHODSThirty-two children diagnosed with ITP between May and August, 2015 and 22 healthy children were enrolled. Flow cytometry was used to determine the Th17/Treg ratio in peripheral blood of healthy children and children with ITP before and after treatment with immunoglobulin.

RESULTSCompared with the patients with ITP before treatment, the healthy children and the patients treated with immunoglobulin had a significantly lower percentage of Th17 cells in CD4+ T cells, a significantly lower Th17/Treg ratio, and a significantly higher percentage of Treg cells in CD4+ T cells in peripheral blood (P<0.05). In the 32 ITP children treated with immunoglobulin, 20 had complete response, 4 had response, and 8 had no response. The patients with complete response had a significantly lower percentage of Th17 cells in CD4+ T cells and a significantly lower Th17/Treg ratio in peripheral blood than the patients without response (P<0.05).

CONCLUSIONSThe Th17/Treg imbalance can be found in children with ITP. Immunoglobulin can improve the cellular immune function by regulation of the Th17/Treg ratio. The Th17/Treg ratio may serve as an indicator for assessing the therapeutic effects of ITP.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

Aged ; Atrial Fibrillation ; surgery ; Catheter Ablation ; methods ; Humans ; Male

OBJECTIVETo determine the changes of CD4+CD25+ regulatory T cells and the levels of IL-10 and TGF-beta1 in the mouse model of asthma and the effects of glucocorticoid inhalation on CD4+CD25+ regulatory T cells and IL-10 and TGF-beta1 levels.

METHODSThirty BALB/c mice were randomly divided into three groups: asthma model, glucocorticoid treatment and control. Asthma was induced by OVA administration in the asthma model and the glucocorticoid treatment groups. The glucocorticoid treatment group received budesonide inhalation (0.8 mg/L) before the challenge of asthma. After 24 hrs of the last challenge, the lung tissues of middle lobe of the right lung were obtained for the observation of lung pathological changes. Peripheral anticoagulated blood and lung lymph cells suspension were collected. The percentage of CD4+CD25+ regulatory T cells in CD4+ T cells was detected by flow cytometry, and the levels of IL-10 and TGF-beta1 in plasma were measured using ELISA.

RESULTSThe percentage of CD4+CD25+ regulatory T cells in peripheral blood and lung lymph cells suspension in the asthma model group was significantly lower than the control group ( P<0.01). The glucocorticoid-treated asthma group showed an increased percentage of CD4+CD25+ regulatory T cells compared with the asthma model group (P<0.01) and a similar percentage of CD4+CD25+ regulatory T cells in peripheral blood and lung lymph cells suspension to the control group. Compared with the control group, plasma levels of IL-10 and TGF-beta1 decreased significantly in the asthma model group (P<0.01). After glucocorticoid treatment plasma IL-10 level increased significantly (P<0.01) and was similar to that in the control group, while plasma TGF-beta1 level remained lower than that in the control group.

CONCLUSIONSThe percentage of CD4+CD25+ regulatory T cells and the levels of IL-10 and TGF-beta1 decreased in asthmatic mice which might contribute to the pathogenesis of asthma. Glucocorticoid can increase the percentage of CD4+CD25+ regulatory T cells and the levels of IL-10 and thus provides protective effects against asthma. The changes of the percentage of CD4+CD25+ regulatory T cells in peripheral blood were consistent with those in the lung, suggesting that monitoring the changes of CD4+CD25+ regulatory T cells in peripheral blood may be useful to understand the changes of CD4+CD25+ regulatory T cells in the lung.

Animals ; Asthma ; drug therapy ; immunology ; Disease Models, Animal ; Female ; Glucocorticoids ; pharmacology ; Interleukin-10 ; blood ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Regulatory ; drug effects ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo compare two different methods, namely orthotopic implantation and armpit implantation, for establishing nude mouse model bearing osteosarcoma in light of the tumor formation rate and time, tumor growth changes and lung metastasis.

METHODSOsteosarcoma cells were inoculated in the left armpit or the femoral medullary canal of nude mice, and the tumor growth was observed 6 weeks later. The differences in the pulmonary metastasis were examined macroscopically and microscopically.

RESULTSArmpit implantation allowed easy operation and fast tumor growth, but tumors often sustained damages by frictions on the surface (P<0.05), and avascular necrosis occurred earlier in the tumors (P<0.05). With this approach, the pulmonary metastasis rate was only 40% (P<0.05), and the metastases in the lungs were sparser (P<0.05) and small. Orthotopic transplantation could well simulate the original growth environment of osteosarcoma, and resulted in milder avascular necrosis and greater pulmonary metastasis rate (100%) with larger and denser metastatic foci. But orthotopic transplantation required more complicated operation for establishing osteosarcoma-bearing models.

CONCLUSIONThe two methods both result in high tumor growth rate, but orthotopic transplantation is superior in inducing pulmonary metastasis of osteosarcoma and reducing friction injury of the tumor and avascular necrosis in the tumor.

Animals ; Bone Neoplasms ; Disease Models, Animal ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; methods ; Osteosarcoma