Objective To analyze the protein expression and subcellular distribution of mechanogrowth factor (MGF) in ostcoblasts under stretch stimulation. Methods Cyclic stretching was applied to osteohlasts by a mechanical stretching device. The whole-cell proteins were extracted from controlled and stretched osteoblasts for detecting the protcin expression level of MGF by Western blot and observing the intracellular distribution of MGF by fluorescent immunocytological method. Results Western blot showed significant increase of expression of MGF in osteoblasts under stimulation of cyclic stretching. The level of protein was increased by four folds after 12-hour stretching of osteohlasts, and then declined sharply. Immunofluorescence analysis showed that MGF was mainly distributed in the nuclei of osteoblasts. ConcinsionsUnder the cyclic stimulation, the expression of MGF reaches a short period of peak in osteoblasts, which may be related to the injury of osteoblasts caused by stretching. MGF is mainly distributed in the nuclei of osteoblasts, indicating that MGF may contain nuclear localization signal and modulate the expression of relative genes.

Objective To investigate the changes of expression of peroxisome proliferator-activated receptor γ (PPARγ) and its coregulators and monocyte chemotactic factor (MCP-1) treated with intedeukin-1β (IL-1β), and to analyze the mechanism of interaction of these factors. Methods Renal tubular cells (HK-2 cells) were cultured in vitro. Total cellular RNA was isolated for real-lime quantitative polymerase chain reaction (real-time PCR), nuclear extracts were prepared for Western blot analysis and EMSA. The supernatant was collected for ELISA after the treatment of IL-1β at different concentrations and time points. Results Under stimulus of different concentrations of IL-1β (0～20 μg/L) for 24 hours, the mRNA expression of PPARγ, SRC-1, SRC-2 and PGC-1 decreased significantly (P<0.05), meanwhile NCoR increased obviously (P<0.05). In further time-dependent experiment, the mRNA levels of SRC-2 and PGC-1 decreased by 57% and 48%, respectively, at 1 hour after treatment with 10 μg/L IL-1β (P<0.05). The expression of SRC-1 decreased by 43%only after 2 hours (P<0.05). The expression of NCoR was not obviously changed until stimulated by IL-1β for 8 hours (2.17 folds, P<0.05), then it decreased slowly. In the same time-dependent experiment, Western blot analysis showed that IL-1β (10 μg/L) significantly decreased the protein level of PPARγ at 4 hours (P<0.05). ELISA analysis revealed that the secretion of MCP-1 kept on rising and reached the peak (160.56±2.80) ng/L at 8 hours (P<0.01), then decreased to (50.82±1.25) ng/L at 24 hours (P<0.01). IL-1β could down-regulate the DNA binding activity of PPARγ, and the activity of NF-κB was up-regulated. Conclusions PPARγ and its eoregulators are closely related to MCP-1 and NF-κB during inflammation response in kidney. The activation of NF-κB by IL-1β leads to the decrease of PPARγ, and its coactivators expression levels, however the expression of MCP-1 and NCoR in renal tubular epithelial cells is up-regulated. PPARγ together with its coregulators participate in the inflammation response in kidney.

ObjectiveTo investigate the role of MG-132, a specific dipeptide proteasome inhibitor, on the proliferation, apoptosis and the related proteins in renal interstitial fibroblasts. MethodsRenal interstitial fibroblasts (NRK-49F) were induced by transforming growth factor β1 (TGF-β1, 5 μg/L) and pro-treated with MG-132 (0～5 μmol/L). The cell proliferation was measured with MTT method. Cell cycle and apoptosis were analyzed by flow cytometry. The apoptosis was also analyzed by Annexin V/PI staining and DNA ladder. Expression of p53, p27, p21, caspase-3, Bcl-2 and Bax protein was examined by Western blot. ResultsTGF-β1 (5 μg/L) could stimulate the proliferation of NRK-49F. MG-132 (0.25～5 μmol/L) could inhibit TGF-β1-induced proliferation in a dose-dependent manner through G1-arrest. TGF-β1 alone could not induce apoptosis (3.880%±0.365% vs 4.723%±1.582%). But pretreatment of MG-132 (0.1～2.5 μmol/L) could significantly induce apoptosis of TGF-β1-stimulated NRK-49F in a dose-dependent manner. Typical DNA ladder was also confirmed in these two groups in the DNA fragments analysis after being incubated with 2.5 μmol/L MG-132 with or without 5 μg/L TGF-β1. Western blot showed that MG-132 could activate the cell-cycle and apoptosis-related proteins such as p53, p21, caspase-3, Bax and inhibit Bcl-2 in a dose-dependent manner, while expression of p27 remained unchanged. ConclusionsProteasome inhibitor MG-132 can inhibit proliferation and induce the cell apoptosis in renal interstitial fibroblasts stimulated by TGF-β1. The mechanism may be associated to the mediation of p53, p21, caspase-3, Bcl-2 and bax pathways. Protoasome inhibitor may be a new strategy to treat renal interstitial fibrosis.

OBJECTIVETo study the influence of Aconiti Lateralis Radix Praeparata on asthenia cold syndrome rats with whole genome gene expression of liver by gene chip technique.

METHODThe asthenia cold syndrome rat models were established by administering traditional Chinese medicine raw Gypsum Fibrosum, Gentianae Radix, Phellodendri Chinensis Cortex and Anemarrhenae Rhizoma. After treated with Aconiti Lateralis Radix Praeparata, the rats' liver gene expressions were detected using gene chip. Differential expression genes were screened for gene function annotation, and some genes were selected to check the accuracy of the results by RT-PCR.

RESULTCompared with the asthenia cold model group, the asthenia cold treatment group showed 212 differential expression genes, mainly involving function of immune response and oxidoreductase activity.

CONCLUSIONAconiti Lateralis Radix Praeparata is proved to have an effect on up-regulating immune response-related genes and oxidizing oxidoreductase activity-related genes of asthenia cold syndrome rats and may be a molecular mechanism for classical warm-nature medicine Aconiti Lateralis Radix Praeparata in warming meridians and dissipating cold.

Aconitum ; chemistry ; Animals ; Asthenia ; drug therapy ; genetics ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Profiling ; Genomics ; Liver ; drug effects ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Wistar

Objective:To investigate the expression levels and clinical significance of glioma-associated oncogene homolog 1 (GLI1) and sonic hedgehog signaling molecule (Shh) in the malignant transformation of ovarian endometriosis (EM).Methods:The expressions of GLI1 and Shh were detected by real-time reverse transcription (RT)-polymerase chain reaction (PCR) and EnVision method in 50 cases of ovarian EM tissues, 35 cases of atypical endometriosis (aEM) and 50 cases of endometriosis-associated ovarian cancer (EAOC). The expression differences of two molecular markers in the malignant transformation of ovarian EM were compared, and the relationships between two molecular markers and the clinicopathological features and prognosis of EAOC were analyzed.Results:(1) RT-PCR showed that the expression levels of GLI1 mRNA in EM, aEM and EAOC group were 1.77±0.40, 3.54±0.44, and 7.80±0.24, respectively. The expression levels of Shh mRNA were 0.95±0.21, 3.14±0.35, and 5.41±0.31, respectively. GLI1 and Shh mRNA in EAOC group were significantly higher than those in EM and aEM group (all P<0.01), and there were statistically significant differences between EM and aEM group (all P<0.01). The percentages of GLI1 in ovarian EM, aEM and EAOC were 32% (16/50), 57% (20/35), and 66% (33/50), respectively, meanwhile, the positive expression rates of Shh were 20% (10/50), 49% (17/35), and 54% (27/50), respectively (all P<0.01). GLI1 mRNA expression was positively correlated with Shh mRNA expression in EAOC tissues ( r=0.721, P<0.01). The expressions of GLI1 protein were proportionated to Shh protein in EAOC tissues ( r=0.608, P=0.001). (2) The expression of GLI1 was significantly related to the International Federation of Gynecology and Obstetrics (FIGO) stage, cancer antigen 125 (CA 125) levels, lymph node metastasis, and Platinum resistance in EAOC patients (all P<0.05). The expression of Shh were related to FIGO stage and lymph node metastasis in EAOC patients (all P<0.05). Logistic regression analysis showed that GLI1 expression was an independent risk factor for poor prognosis in EAOC patients ( P<0.05). Kaplan-meier survival analysis showed that the overall survival rate of EAOC patients with high GLI1 expression and low GLI1 expression was 12.1% and 35.3%, respectively, with statistical significance ( χ2=10.73, P<0.01). The overall survival rate of EAOC patients with high and low expression of Shh protein was 11.1% and 30.4%, in which there was statistically significant difference ( χ2=3.96, P=0.047). Conclusion:GLI1 and Shh are highly associated with the malignant transformation of ovarian EM, which may play a role in promoting malignant degeneration of ovarian EM, and the high expression of GLI1 and Shh indicates a poor prognosis in EAOC patients.

Objective To explore the regulatory role of calcium activated chloride channel (CaCC) in vascular structural remodeling in pathogenesis of pulmonary arterial hypertension (PAH) induced by high pulmonary blood flow.Method An abdominal aorta and inferior vena cava shunting operation was used to induce high pulmonary blood flow and establish a PAH rat model.Seventy-five SD rats were randomly divided into normal,sham,shunt,niflumic acid (NFA) 1 (0.2 mg/(kg · d)) and NFA 2 (0.4 mg/(kg · d)) groups.There were 15 rats in each group.Pulmonary artery pressure and vascular structural remodeling were measured,arteriole contraction ratio among these groups were compared using vascular tone analysis system,and the electrophysiology of pulmonary artery smooth muscle cell (PASMC) was recorded using patch clamp technology.Differences between multiple groups were compared through variance analysis and that between groups with q test.Result Compared with normal ((14.4 ± 1.3) mmHg,1 mmHg =0.133 kPa) and sham groups ((13.5 ± 2.3) mmHg),mean pulmonary artery pressure in shunt group ((27.4 ± 2.4) mmHg) increased significantly (P ＜ 0.05).Compared with shunt group,mean pulmonary artery pressure in NFA 1 group ((21.2 ± 2.0) mmHg) and NFA 2 group ((22.3 ± 2.0) mmHg) decreased significantly (P ＜ 0.05).Pulmonary vascular structural remodeling including pulmonary artery stenosis presented in shunt group.Compared with normal ((114.3 ± 1.2) ％) and sham ((115.5 ± 1.1) ％) groups,arteriole contraction ratio to 10-5 mol/L phenylephrine in shunt group ((132.6 ± 1.4) ％) increased significantly (P ＜ 0.05).Compared with shunt group,pulmonary vascular structural remodeling alleviated in NFA 1 and NFA 2 groups.Arteriole contraction ratio in NFA 1 group ((126.4 ± 1.3) ％) and NFA 2 group ((124.6 ± 1.0) ％) decreased significantly compared with shunt group (P ＜ 0.05).Patch clamp technique recorded typical CaCC currents.Compared with normal ((32.3 ±2.3) pA/pF) and sham groups ((35.3 ± 1.2) pA/pF),the CaCC current density of PASMC in shunt group ((51.3 ± 2.7) pA/pF) increased significantly (P ＜ 0.05).Compared with shunt group,the CaCC current density of PASMC in NFA 1 group ((40.2 ± 1.5) pA/pF) and NFA 2 group ((42.7 ± 2.2) pA/pF) decreased significantly (P ＜ 0.05).Conclusion CaCC is involved in pulmonary arterial hypertension induced by high pulmonary blood flow through regulating membrane potential.NFA attenuate pulmonary vascular structural remodeling and pulmonarv pressure through decreasing CaCC current density of PASMC membrane.

In order to ensure normal body function, the human body is dependent on a tight control of its blood glucose levels. This is accomplished by a highly sophisticated network of various hormones and neuropeptides released mainly from the brain, pancreas, liver, intestine as well as adipose and muscle tissue. Within this network, the pancreas represents a key player by secreting the blood sugar-lowering hormone insulin and its opponent glucagon. However, disturbances in the interplay of the hormones and peptides involved may lead to metabolic disorders such as type 2 diabetes mellitus (T2DM) whose prevalence, comorbidities and medical costs take on a dramatic scale. Therefore, it is of utmost importance to uncover and understand the mechanisms underlying the various interactions to improve existing anti-diabetic therapies and drugs on the one hand and to develop new therapeutic approaches on the other. This review summarizes the interplay of the pancreas with various other organs and tissues that maintain glucose homeostasis. Furthermore, anti-diabetic drugs and their impact on signaling pathways underlying the network will be discussed.
Blood Glucose ; Brain ; Comorbidity ; Diabetes Mellitus, Type 2 ; Glucagon ; Glucose* ; Hand ; Homeostasis* ; Human Body ; Insulin ; Intestines ; Liver ; Neuropeptides ; Pancreas ; Peptides ; Prevalence

Objective:To investigate the relationship between the expression level of platelet microparticle (PMP) and the disease activity of inflammatory bowel disease (IBD) in IBD patients, and to explore the ability of PMP from different sources to induce the formation of neutrophil extracellular trap (NET) in vitro. Methods:From May 2018 to July 2019, 118 patients with IBD admitted to the Department of Gastroenterology at The First Affiliated Hospital of Anhui Medical University were selected, among whom 54 cases were ulcerative colitis (UC) patients (UC group) and mild, moderate and severe cases were 17, 25 and 12, respectively; and 64 cases were Crohn′s disease (CD) (CD group), 6 were in remission stage, and mild, moderate, severe cases were 27, 22 and 9, respectively. During the same period, 35 healthy individuals with normal checkups were selected as the healthy control group. Specimens were collected and the expression levels of PMP were measured by flow cytometry.And the correlation between the expression level of PMP and the disease activity index (DAI) score was analyzed.NET formation experiment groups were set up, including neutrophils of healthy control group (6 cases), neutrophils of IBD group (6 cases), neutrophils of healthy controls + PMP of IBD group (12 cases) and neutrophils of healthy controls+ PMP group (6 cases). After immunofluorescence staining, the proportion of NET formation of each group was observed under laser scanning confocal microscopy (LSCM). Mann-Whitney U test, Spearman correlation analysis and Independent-sample t test were used for statistical analysis. Results:The expression levels of PMP in peripheral blood of the UC group and the CD group were 2 184.5(2 817.0)/μL and 2 209.0(2 409.0)/μL, respectively, which were all higher than that of the healthy control group (776.0(407.0)/μL), and the differences were statistically significant ( U=-6.018 and -6.426, both P<0.01). The expression level of PMP of patients with severe UC was 3 873.0(4 611.3)/μL, which was higher than those of patients with mild or moderate UC (1 248.0(1 888.0)/μL and 1 432.0(1 783.0)/μL, respectively), and the differences were statistically significant ( U=-2.745 and -2.547, both P<0.05). The expression level of PMP of patients with severe CD was 5 658.0(5 067.5)/μL, which was higher than those of patients with mild or moderate CD or in remission (1 327.5(1 934.0)/μL, 1 405.0(2 965.0)/μL and 2 300.0(1 552.0)/μL, respectively), and the differences were statistically significant ( U=-1.650, -1.955 and -1.306, all P<0.05). There was no statistically significant difference in the expression level of PMP between the UC group and the CD group, between the mild and moderate UC patients, and between the CD in remission and the mild, moderate patients (all P>0.05). The results of correlation analysis showed that the expression levels of PMP in peripheral blood of patients with UC or CD were positively correlated with DAI score and CRP ( r=0.406, 0.358, 0.325, and 0.256; all P<0.05). The proportion of NET formation in the neutrophils of healthy control+ PMP of IBD group was (14.67±5.35) %, which was higher than those of the neutrophils of healthy control groap, neutrophils of IBD group and neutrophils of healthy control+ PMP group ((2.00±0.63)%, (1.67±0.82)% and (5.83±2.86)%), and the differences were statistically significant ( t=5.694, 8.230 and 3.748, all P<0.05). There was no statistically significant difference in the proportion of NET formation between the neutrophils of healthy control group and the neutrophils of IBD group ( P>0.05). Conclusions:The expression level of PMP in peripheral blood of IBD patients increases and is correlated with the disease activity degree in IBD patients. PMP has the ability to induce the NET formation in neutrophils. Moreover, PMP of IBD patients is more likely to induce NET formation than those of healthy individuals, which may be involved in the intestinal inflammatory process by activating neutrophils to produce NET.

Aged ; Bone Marrow ; metabolism ; Cytokines ; metabolism ; Female ; Humans ; Karyotype ; Male ; Mesenchymal Stromal Cells ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism

ObjectiveTo identify the possible health impact of the 14th five⁃year plan for the development of employment and social security in Deqing County and propose improvement measures through health impact assessment. MethodsBased on the data of Deqing County， stakeholder interviews and Delphi Consultation Method， this study described the current status of employment and social security and analyzed the potential health impacts of implementing the 14th five⁃year plan for the development of employment and social security in Deqing County. ResultsThrough a quick assessment process， the results showed that the implementation of the plan would bring mixed health impacts. Positive impacts included enhanced social security capacity， improved health levels of low-income populations and families， increased convenience of medical treatment， and improved efficiency of health services. Negative impacts included reduced accessibility of digital services for the elderly， increased gap in benefits for retirees， increased risk of discrimination against disabled individuals， increased risks of layoffs and unemployment for vulnerable groups， and increased employment instability for middle-aged and elderly populations. ConclusionThe 14th five⁃year plan for the development of employment and social security in Deqing County will bring a series of positive health impacts， but negative health impacts also warrant attention.