Content of osteocyte in bone tissue is the most abundant, the most widely distributed, and embedding the cells in the mineralized bone matrix, the life can be close to the life of the body. Osteocyte formed by the cytoplasm dendritic communication network system between osteocyte and bone surface, is of great significance to maintaining the normal physiological function of bone tissue. Bone cells as the direct receptor of bone mechanical stress, through the release of bioactive factors such as PEG2, NO, ATP and classic Wnt/beta-catenin signaling pathway mechanical stress signal can be converted to bone formation and bone resorption of biochemical signals, and the biochemical signals were transfer to the other type cells of the tissue to regulate its function activities and cause bone reconstruction function. The microcracks surrounding osteocyte can disrupt the microtubule network system,cause surrounding osteocyte autophagy. In addition, osteocyte is very important for regulation of the body mineral balance, fat metabolism, and hematopoietic function.
Animals ; Autophagy ; Hematopoiesis ; Humans ; Minerals ; metabolism ; Osteocytes ; physiology ; Signal Transduction ; Stress, Mechanical

Background Though nitric oxide (NO) and NO synthase (NOS) have a critical role in angiogenesis,their effects on corneal neovascularization (CNV) and mechanism need to be further explored.Objective The aim of this study was to explore the effects of NOS and its antagonist,Nw-nitro-L-arginine methyl ester (L-NAME) on experimental CNV in mice,and investigate the influence of NOS and L-NAME on the tube formation of human retinal endothelial cells (RECs) in vitro.Methods The CNV models were established in the left eyes of 36 male BALB/c mice aged 7-8 weeks by application of the filter paper with NaOH in the center of corneas.The mice were randomized into two groups.L-NAME of 10 g/L (0.5 ml) was intraperitoneally injected 1 week before induction of CNV three times a week for three weeks in the mice of the L-NAME injection,and PBS was used in the same way in the control group.CNV was examined under the slit lamp biomicroscope 2,4,7,14 days after NaOH burn.The expression of CD31 in the CNV was assayed to calculate the ratio of CNV area and total corneal area using whole mount technique.The expression of NOS mRNA in the corneal tissue was detected by reverse transcriptase polymerase chain reaction (PCR),and VEGF expression in the human RECs was assayed by Western blot.The vessel formation number of cultured human RECs with or without L-NAME was performed by matrigel in vitro.Grouped t test was used to compare the differences of the parameters between the two groups.Results CNV developed and peaked 2 weeks after the application of NaOH on the mice corneas,and the CNV was obviously less in the L-NAME group compared with the control group.The expression of NOS mRNA in the corneas (NOS mRNA/ GAPDH mRNA)was significantly lower in the L-NAME group than that of the control group 2,4,7 days after CNV induction (t =19.481,t=22.059,t=10.961,all at P＜0.01).The ratio of the CD31 positive area in whole corneal area was 0.59± 0.01 in the L-NAME group,and that of the control group was 0.78±0.10,showing a significant difference between the two groups (t =3.078,P＜0.05).Western blot assay showed that the relative expression of VEGF protein in human RECs was declined in the L-NAME group compared with the control group 0,2,4,7 days,with statistically significant differences in 4 days and 7 days after NaOH burn(t=7.696,t=17.953,both at P＜0.01).The number of vessel network was 46.33±1.86 in the L-NAME group and 64.00±4.51 in the control group,with a significant difference between them (t =3.623,P＜0.05).Conclusions NOS participated in the pathogenesis and aggravation of CNV induced by NaOH.L-NAME arrests CNV formation and human RECs tube formation through down regulating the VEGF expression and NOS activity.

Objective To study the molecular mechanism of interleukin 25 (IL-25)expression in the lung of asthmatic rats.Methods The expressions of IL-25 mRNA and protein in the lungs were detected by Real-time PCR and ELISA,respectively.The levels of IL-25 mRNA and protein were detected by ovalbumin (OVA)in human bronchial epithelial cells.And the transcription factors that regulate IL-2 5 expression were explored through site prediction.Results The expressions of IL-25 mRNA and protein in the lung of OVA-induced asthma rats were significantly increased during animal experiments.Cell experiments showed that OVA could increase the expression of IL-2 5 in human bronchial epithelial cells in a dose-dependent manner,and OVA could upregulate the expression of transcription factor AP1.AP1 was found in the promoter region of IL-25 by site prediction.The AP1 inhibitor (T5224)significantly reduced the expression of IL-25 in OVA-induced human bronchial epithelial cells. Conclusion The molecular mechanism of IL-25 expression induced by OVA in asthma is related to the increase of transcription factor AP1 .

Objective: To investigate the possible mechanisms in acupuncture analgesia by interaction of &opioid receptor with neurotransmitter transport proteins or the Na+-K+pump. Methods: Microinjection of respective heterologous cRNA into the Xenopus oocytes as a model system, and measurement of steady-state currents under two-electrode voltage clamp. Results: The co-expression of the δ-opioid receptor with GAT1, EAAC1 or the sodium pump resulted in reducing activity of the respective transporter. Opioid receptor activation affected transporter activity in different ways: 1) GAT1 was further inhibited; 2) EAAC1 was stimulated; 3) Na+-K+ pump activity interfered with agonist sensitivity of DOR. Pump inhibition led to higher sensitivity for DPDPE. Conclusion: GABA transporter inhibition and glutamate transporter stimulation may counteract pain sensation by affecting the neurotransmitter concentration in the synaptic cleft and, therefore, may contribute synergistically to pain suppression by acupuncture. Sodium pump inhibition by endogenous ouabain may amplify these effects. These synergistic effects may be the molecular mechanism of inhibiting pain sense and/or acupuncture analgesia.

Abstract: Objective　To explore the spatial epidemiological characteristics of severe cases hand, foot and mouth disease (HFMD) in Guangxi, China, from 2014 to 2018, and to provide a basis for identifying the high-risk regions as well as the prevention and control of severe cases of HFMD in Guangxi. Methods　Spatial-temporal scanning analysis, global and local spatial autocorrelation analysis were used to analyze the spatial clustering of HFMD. The trend surface analysis was used to evaluate the spatial distribution trend of HFMD. Results　From 2014 to 2018, the incidence and severe case fatality rates of HFMD were 3.89/100 000 and 4.23%, respectively. Monte Carlo scanning analysis showed that the first cluster region was Cenxi City, the second cluster was mainly concentrated in northwest of Guangxi, and the aggregation time was mainly concentrated in April to May and August to October. The global spatial autocorrelation analysis showed that the severe HFMD was significant clustering distribution, and the Moran's I coefficients of the sever cases, severe morbidity and severe case fatality rate were 0.088, 0.118, 0.197, respectively (P<0.05). Local spatial autocorrelation analysis showed that hotspots of severe HFMD cases were concentrated in the southern Guangxi, mainly in Lingshan County. Anselin local Moran's I clustering and outlier analysis indicated that 5 high-high (H-H) clustering regions for fatality were Lingshan, Pubei, Zhongshan, Zhaoping and Pinggui County. There were 6 high-high (H-H) clustering regions for severe incidence rate, namely Lingshan, Qinnan, Lingyun, Youjiang, Bama Yao Autonomous and Pinggui County, and 1 high-low (H-L) clustering region, Cenxi County. The trend surface analysis showed that the overall number of severe cases of death decreased from east or west to the middle, and increased from north to middle, and then decreased to south. Conclusions　Severe HFMD cases in Guangxi have obvious spatial-temporal clustering, and the hop spots are mainly concentrated in southern Guangxi. The prevention and control of HFMD in areas with high incidence of severe cases should be strengthened to reduce the burden of HFMD cases.

OBJECTIVETo study the effects of Dihydromyricetin (DMY) on antilipid-peroxidation.

METHODThe antilipid-peroxidation of DMY on heart, liver, brain tissue homogenate and mitochondria was measured by the determination of malondiadehyde (MDA) induced by Fe2+ -Vit C, Fe2+ -H2O2, Fe-Cys with TBA spectrometric method.

RESULTDMY could inhibit the lipid peroxidation of homogenate and mitochondria. The inhibition exhibited concentration-dependent manner.

CONCLUSIONDMY has good antilipid-peroxidation effect, which is worth studing further.

Ampelopsis ; chemistry ; Animals ; Antioxidants ; isolation & purification ; pharmacology ; Brain ; metabolism ; Dose-Response Relationship, Drug ; Flavonols ; isolation & purification ; pharmacology ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; metabolism ; Mitochondrial Swelling ; drug effects ; Myocardium ; metabolism ; Plants, Medicinal ; chemistry ; Rats

OBJECTIVETo investigate the effects of Yougui drink on osteogenic differentiation of bone mesenchymal stem cells (BMSCs) in the proximal femur in steroid-induced avascular necrosis of femoral head in vitro.

METHODSThe BMSCs were induced to osteoblasts in vitro with a conditional medium. The cells cultured with Yougui drink serum were compared with the ordinary one, which both were examined by microscopy, MTU method, ALP and calcium node.

RESULTSMicroscopy and MTU studies showed that Yougui drink could improve proliferation of the osteoblasts. ALP and calcium node studies showed that Yougui drink could improve the activity of the osteoblasts (P < 0.01).

CONCLUSIONYougui drink has positive effects on osteogenic differentiation of mesenchymal stem cells in the proximal femur in steroid-induced avascular necrosis of femoral head in vitro.

Cell Differentiation ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Femur Head Necrosis ; drug therapy ; physiopathology ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Middle Aged ; Osteoblasts ; cytology ; drug effects ; Osteonecrosis ; Steroids ; pharmacology

BACKGROUND: Acute kidney injury following percutaneous coronary intervention (PCI) is associated with a worse outcome. However, the risk factors and outcomes of acute kidney injury (AKI) in patients after intracoronary stent implantation are still unknown. METHODS: A retrospective case control study was done in 325 patients who underwent intracoronary stent implantation from January 2010 to March 2011 at the Drum Tower Hospital of Nanjing University School of Medicine. Those were excluded from the study if they had incomplete clinical data. The patients were divided into a normal group and a AKI group according to the standard of post-operation day 7 to identify AKI. The parameters of the patients included: 1) pre-operative ones: age, gender, hypertension, diabetes mellitus, cerebrovascular disease, left ventricular insufficiency, peripheral angiopathy, creatinine, urea nitrogen, estimated glomerular filtration rate (eGFR), hyperuricemia, proteinuria, emergency operation, hydration, medications (ACEI/ARBs, statins); 2) intraoperative ones: dose of contrast media, operative time, hypotension; and 3) postoperative one: hypotension. The parameters were analyzed with univariate analysis and multivariate logistical regression analysis. RESULTS: Of the 325 patients, 51(15.7％) developed AKI. Hospital day and in-hospital mortality were increased significantly in the AKI-group. Univariate analysis showed that age, pre-operative parameters (left ventricular insufficiency, peripheral angiopathy, creatinine, urea nitrogen, estimated glomerular filtration rate, hyperuricemia, proteinuria, hydration), emergency operation, intraoperative parameters (operative time, hypotension) and postoperative hypotension were significantly different. However, multivariate logistic regression analysis revealed that increased age (OR=0.253, 95％CI=0.088–0.727), pre-operative proteinuria (OR=5.351, 95％CI=2.128–13.459), pre-operative left ventricular insufficiency (OR=8.704, 95％CI=3.170–23.898), eGFR≤60 ml/min/1.73 m2 (OR=6.677, 95％CI=1.167–38.193), prolonged operative time, intraoperative hypotension (OR=25.245, 95％CI=1.001–1.034) were independent risk factors of AKI. CONCLUSIONS: AKI is a common complication and associated with ominous outcome following intracoronary stent implantation. Increased age, pre-operative proteinuria, pre-operative left ventricular insufficiency, pre-operative low estimated glomerular filtration rate, prolonged operative time, intraoperative hypotension were the significant risk factors of AKI.

OBJECTIVETo investigate the effect of L-arginine on expression of human FVIII gene.

METHODSPlasmid pcDNA6/V5-HisA-BDDhF VIII containing B domain deleted human coagulant factor VIII cDNA (BDDhF VIII cDNA) was constructed and transfected into human umbilical vein endothelial cells (HUVEC). After 72 h incubation with L-arginine (final concentration was 10 mmol/L) , the supernatant was collected for determining the antigen and clotting activity of human FVIII (FVIII: Ag and FVIII: C ) with ELISA and one stage clotting assay respectively. HUVECs were harvested for detecting human FVIII mRNA by Northern blot analysis. The five functional domains of BDDhFVIII cDNA including A1, A2, A3, C1 and C2 were amplified with PCR and inserted into pcDNA6/V5-HisA to construct the expression plasmids pcDNA6/V5-Hi-sA-BDDhFVIII-A1, pcDNA6/V5-HisA-BDDhFVIII-A2, pcDNA6/V5-HisA-BDDhFVIII-A3, pcDNA6/V5-HisA-BDDhFVIII-C1 and pcDNA6/V5-HisA-BDDhFVIII-C2, respectively. HUVEC were transfected with the five plasmids respectively and incubated with L-arginine (at the final concentration of 10 mmol/L) for 72 h. Nucleoli were then isolated and underwent run-on assay.

RESULTSAfter 24 h incubation with L-arginine, FVIII: Ag and FVIII: C were increased markedly in the supernatant of HUVEC [FVIII: Ag was (146.08 +/- 4.78) ng/ ml, and FVIII: C (0.752 +/- 0.009) U/ml/10(6) cells x 24 h, while in control supernatant without L-arginine, FVIII: Ag was (34.66 +/- 3.98) ng/ml, and FVIII: C (0.171 +/- 0.006) U/ml/10(6) cell x 24 h, P < 0.01]. Northern blot analysis indicated that, after adding L-arginine, the transcription of human FVIII mRNA was intensified remarkably in HUVEC transfected with pcDNA6/V5-HisA-BDDhFVIII, but no any transcription in those transfected with pcDNA6/V5-HisA. Run-on assay demonstrated that with L-arginine induction, A1 and A2 domains transcription was increased obviously, while no change in A3, C1 and C2 domains transcription.

CONCLUSIONL-arginine increases expression of human FVIII gene in HUVEC through enhancing its transcription, particularly, domain A1 and A2 within FVIII gene.

Arginine ; pharmacology ; Cells, Cultured ; DNA, Complementary ; genetics ; Endothelial Cells ; metabolism ; Factor VIII ; genetics ; Gene Expression ; drug effects ; Genetic Vectors ; Humans ; Plasmids ; genetics ; Transcription, Genetic ; drug effects ; Transfection ; Umbilical Veins ; cytology

The study is to investigate the effect of angiotensin II (Ang II) and its receptor blockers on migration and endothelin-1 (ET-1) expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang II significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. Ang II (1 x 10(-8) - 1 x 10(-6) mol x L(-1)) significantly increased the expression of preproET-1 mRNA in round cells (P < 0.01), and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression of preproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. The effects of Ang II and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression of preproET-1 mRNA. The results indicate that there are two cell subpopulations: round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative process.
Angiotensin II ; pharmacology ; Angiotensin Receptor Antagonists ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Endothelin-1 ; genetics ; metabolism ; Fibroblasts ; cytology ; metabolism ; Imidazoles ; pharmacology ; Losartan ; pharmacology ; Male ; Pyridines ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Vasoconstrictor Agents ; pharmacology