8-Azaguanine-induced autophagy contributes to its chemoresistance in hepatic cancer cells / 药学学报

Autophagy, an evolutionarily conserved process by which components of the cell are degraded in lysosomes, may facilitate survival of cancer cells under stress conditions. 8-Azaguanine (8-AG), an inhibitor of purine nucleotide biosynthesis, shows antineoplastic activity in multiple tumor cells. However, chemoresistance has restricted its development as an anticancer agent, and the mechanism of 8-AG resistance is not fully understood. We report here that 8-AG induces a protective autophagy to eliminate its cytotoxicity, and inhibition of autophagy increases cellular sensitivity of cancer cells to 8-AG treatment. Using HepG2 or SMMC-7721 hepatic cancer cell lines, we found that 8-AG inhibited cell viability and induced intrinsic apoptosis, accompanied by the up-regulation of the pro-apoptotic protein BimS, one of Bim (also known as BCL-2-like protein 11, BCL2L11) isoforms. Furthermore, 8-AG treatment enhanced the autophagy flux by promoting the dephosphorylation and activation of Unc-51-like autophagy activating kinase 1 (ULK1) via Akt/mTORC1 (mammalian target of rapamycin complex 1) signaling inhibition. Depletion of autophagy-related gene 7 (ATG7) markedly enhanced the level of BimS, and promoted cell death in response to 8-AG. 8-AG in combination with autophagy inhibitor chloroquine (CQ) or bafilomycin A1 (Baf A1) promoted the 8-AG-induced apoptosis in hepatic cancer cells. Altogether, these findings suggest that autophagy promotes chemoresistance of cancer cells for 8-AG, and blocking autophagy increases cellular sensitivity of cancer cells to 8-AG treatment.

Association of TLR1 gene polymorphisms with pulmonary candida infection and candida colonization in Guizhou Han patients with chronic obstructive pulmonary disease / 医学研究生学报

Objective Studies are rarely reported on the correlation of the Toll-like receptor (TLR) gene polymorphisms with fungal infection in the Chinese Han population. This study aims to explore the association between TLR gene polymorphisms and pulmonary candida infection (PCI) and candida colonization (CC) in patients with chronic obstructive pulmonary disease (COPD) in the Guizhou Han population. Methods Using the polymerase chain reaction-direct sequencing (PCR-SBT) method, we genotyped six single nucleotide polymorphisms (SNP) of the TLR1, TLR2 and TLR4 genes in 344 Guizhou Han patients with COPD, including 80 cases complicated by PCI (the PCI group), 103 cases complicated by CC (the CC group), and 161 negative controls. We analyzed the correlation of the SNPs with PCI and CC in the COPD patients in different genetic models with the SNPstats online software, measured the levels of IL-6, IL-8, IL-1β and TNF-α proteins in the plasma of the patients by ELISA, and assessed the functional consequences of these polymorphisms. Results The polymorphisms of rs5743611, rs5743708, rs4986790 and rs4986791 were found in none of the patients. The genotype frequency of rs4833095 was significantly different between the PCI and control groups in the codominant, dominant and overdominant models (P < 0.05), with the dominant model as the best genetic pattern. No statistically significant difference was observed either in the rs4833095 genotype frequency between the CC and control groups (P > 0.05) or in the rs5743618 genotype frequency between any two groups (P > 0.05). In the PCI group, the T allele of the rs4833095 polymorphism significantly decreased the levels of IL-6, IL-8 and IL-1β in the plasma (P < 0.05), and that of TNF-α as well, though with no statistically significant difference (P > 0.05). Conclusion The rs4833095 polymorphism of the TLR1 gene is associated with PCI in COPD patients. The T allele of rs4833095 may affect the function of TLR1, decrease the levels of IL-6, IL-8 and IL-1β proteins in the plasma. Neither rs4833095 nor rs5743618 gene polymorphism is correlated with the susceptibility to candida colonization.

Early cardiac injury in patients with obstructive sleep apnea / 中国应用生理学杂志

OBJECTIVE@#To evaluate the early cardiac injury caused by obstructive sleep apnea (OSA) before the development of cardiovascular symptoms of OSA.@*METHODS@#Ninety-two patients without any known cardiovascular disorders who underwent polysomnography (PSG) were enrolled in the study. Subjects were divided into mild, moderate, and severe OSA groups by their apnea hypopnea index (AHI), and 25 healthy individuals were identified as controls. After PSG examination, fasting blood samples for the evaluation of N-terminal pro-brain natriuretic peptide (NT-proBNP) and heart-type fatty acid binding protein (h-FABP) were collected in the morning, and left ventricular(LV) functions were assessed by using echocardiographic methods. Thirty moderate and severe OSA patients were treated with continuous positive airway pressure respectively (CPAP).@*RESULTS@#The levels of h-FABP and NT-proBNP were obviously higher in all OSA groups than those in the control group (<0.01), and were positively correlated with AHI (<0.01). The Em/Am values of all OSA groups and E/A values of the moderate and severe OSA groups were significantly reduced (<0.01). The difference in Em/Am values among the groups was statistically significant (<0.01). Compared with those before treatment, h-FABP and NT-BNP levels in serum of OSA patients after CPAP treatment were significantly reduced (<0.01), and Em/Am and E/A values were significantly increased (<0.01).@*CONCLUSIONS@#Left ventricular diastolic dysfunction and early myocardial microtrauma are major manifestations of early heart damage in patients with OSA. CPAP therapy could significantly improve early cardiac damage in OSA patients.

Heparin-derived oligosaccharide inhibits vascular intimal hyperplasia in balloon-injured carotid artery / 中国天然药物

The aims of the present study were to determine the effects of heparin-derived oligosaccharides (HDOs) on vascular intimal hyperplasia (IH) in balloon-injured carotid artery and to elucidate the underlying mechanisms of action. An animal model was established by rubbing the endothelia within the common carotid artery (CCA) in male rabbits. The rabbits were fed a high-cholesterol diet. Arterial IH was determined by histopathological changes to the CCA. Serum lipids were detected using an automated biochemical analysis. Expressions of mRNAs for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1), scavenger receptor class B type I (SR-BI), and ATP-binding cassette transporter A1 (ABCA-1) were analyzed using reverse transcription polymerase chain reaction assays. Expressions of VEGF, VCAM-1, MCP-1, SR-BI and ABCA-1 proteins were analyzed by Western blotting. Enzyme-linked immunosorbent assays were used to quantify expression levels of VEGF and bFGF. Our results showed that administration of HDO significantly inhibited CCA histopathology and restenosis induced by balloon injury. The treatment with HDOs significantly decreased the mRNA and protein expression levels of VEGF, bFGF, VCAM-1, MCP-1, and SR-BI in the arterial wall; however, ABCA-1 expression level was elevated. HDO treatment led to a reduction in serum lipids (total cholesterol, triglycerides, high-density and low-density lipoproteins). Our results from the rabbit model indicated that HDOs could ameliorate IH and underlying mechanism might involve VEGF, bFGF, VCAM-1, MCP-1, SR-BI, and ABCA-1.

Effect of PPARγ agonist (rosiglitazone) on the secretion of Th2 cytokine in asthma mice

OBJECTIVE@#To explore the effect of PPARγ agonist (rosiglitazone) on the secretion of Th2 cytokines and the proportion of immune cell subsets in asthma mice.@*METHODS@#Ovalbumin (OVA)-sensitized mice were used to build asthma models. Those mice were divided into the normal control group, model group and rosiglitazone group. Differences of the changes in lung histopathology of mice in the three groups were observed through hematoxylin and eosin (HE) strain, and the numbers of the total cells, eosinophils and neutrophils in BALF of mice in the three groups were compared. ELISA and real-time PCR were employed to detect the protein levels of interleukin (IL)-5, IL-13, IL-4 and IL-10 and mRNA level, respectively. Flow cytometry number was implied to analyze the proportion of immune cell subsets in peripheral blood of mice.@*RESULTS@#Compared with the mice in the control group, and mice of the model group, the infiltration of inflammatory cells in BALF increased, bronchial smooth muscle became thickened, a large amount of collagen deposited, the secretion of Th2 cytokine increased significantly, the ratio of regulatory T cells (Treg) decreased, the ratio of T17 cells rose distinctly; while in mice of the rosiglitazone group, the changes of their lung histopathology were improved obviously, the number of infiltration of inflammatory cells declined, the thickened smooth muscle relieved, the deposition of collagen decreased, the secretion of Th2 cytokine was inhibited, the ratio of Treg went up, and the increased of the ratio of T17 cells was inhibited but still not return to normal level.@*CONCLUSIONS@#Rosiglitazone can regulate the proportion of Treg and Th17 cells and inhibit the secretion of Th2 cytokines, which inhibit the airway inflammatory response for asthma mice effectively.

Research in progress on natural polycyclic polyprenylated acylphoroglucinols / 中草药

As major active components of the plant of Guttiferae family, polycyclic polyprenylated acylphoroglucinols (PPAPs) have become a hot topic in natural product studies for their novel structures and varied bioactivities such as antibacterial, antiviral, antitumor, and antidepressant activities. In this paper, the chemical structures and bioactivities of 226 natural PPAPs have been summarized. It may provide the reference for the further studies of PPAPs.

Effect of PPARγ agonist (rosiglitazone) on the secretion of Th2 cytokine in asthma mice

Objective To explore the effect of PPARγ agonist (rosiglitazone) on the secretion of Th2 cytokines and the proportion of immune cell subsets in asthma mice. Methods Ovalbumin (OVA)-sensitized mice were used to build asthma models. Those mice were divided into the normal control group, model group and rosiglitazone group. Differences of the changes in lung histopathology of mice in the three groups were observed through hematoxylin and eosin (HE) strain, and the numbers of the total cells, eosinophils and neutrophils in BALF of mice in the three groups were compared. ELISA and real-time PCR were employed to detect the protein levels of interleukin (IL)-5, IL-13, IL-4 and IL-10 and mRNA level, respectively. Flow cytometry number was implied to analyze the proportion of immune cell subsets in peripheral blood of mice. Results Compared with the mice in the control group, and mice of the model group, the infiltration of inflammatory cells in BALF increased, bronchial smooth muscle became thickened, a large amount of collagen deposited, the secretion of Th2 cytokine increased significantly, the ratio of regulatory T cells (Treg) decreased, the ratio of T17 cells rose distinctly; while in mice of the rosiglitazone group, the changes of their lung histopathology were improved obviously, the number of infiltration of inflammatory cells declined, the thickened smooth muscle relieved, the deposition of collagen decreased, the secretion of Th2 cytokine was inhibited, the ratio of Treg went up, and the increased of the ratio of T17 cells was inhibited but still not return to normal level. Conclusions Rosiglitazone can regulate the proportion of Treg and Th17 cells and inhibit the secretion of Th2 cytokines, which inhibit the airway inflammatory response for asthma mice effectively.

Expressions of Mast Cell Tryptase and Brain Natriuretic Peptide in Myocardium of Sudden Death due to Hypersensitivity and Coronary Atherosclerotic Heart Disease / 法医学杂志

OBJECTIVES@#To explore the value of mast cell tryptase and brain natriuretic peptide（BNP） in the differential diagnostic of sudden death due to hypersensitivity and coronary atherosclerotic heart disease.@*METHODS@#Totally 30 myocardial samples were collected from the autopsy cases in the Department of Forensic Pathology, Shanxi Medical University during 2010-2015. All samples were divided into three groups： death of craniocerebral injury group, sudden death of hypersensitivity group and sudden death of coronary atherosclerotic heart disease group, 10 cases in each group. Mast cell tryptase and BNP in myocardium were detected by immunofluorescence staining and Western Blotting.@*RESULTS@#Immunofluorescence staining showed that the positive staining mast cell tryptase appeared in myocardium of sudden death of hypersensitivity group and coronary atherosclerotic heart disease group. Among the three groups, the expression of mast cell tryptase showed significantly differences through pairwise comparison （P<0.05）; The expression level of BNP in sudden death of coronary atherosclerotic heart disease group were significantly higher than the sudden death of hypersensitivity group and death of craniocerebral injury group （P<0.05）. The difference of the expression level of BNP between the sudden death of hypersensitivity group and the death of craniocerebral injury group had no statistical significance （P>0.05）.@*CONCLUSIONS@#The combined detection of the mast cell tryptase and BNP in myocardium is expected to provide help for the forensic differential diagnosis of sudden death due to hypersensitivity and coronary atherosclerotic heart disease.

Anti-inflammatory role of microRNA let-7c in LPS treated alveolar macrophages by targeting STAT3

OBJECTIVE@#To explore the expression of microRNA (miRNA) let-7c and its function in chronic obstructive pulmonary disease (COPD) and alveolar macrophage cells.@*METHODS@#Real time PCR was performed to detect the expression of miRNA let-7c in the lung tissue of COPD patients and COPD model in mice. MiRNA let-7c was overexpressed in alveolar macrophages isolated from mice and its effect was measured by the production of pro-inflammation cytokines and the protein level of signal transducer and activator of transcription 3 (STAT3) as well as phosphorylation level of STAT3 after LPS stimulation. Luciferase assay was used to detect the binding of miRNA let-7c and 3'UTR of STAT3.@*RESULTS@#MiRNA let-7c expression was significantly lower in patients with COPD compared with control group, and the similar result was found in COPD mice and LPS stimulated alveolar macrophages. Overexpression of miRNA let-7c in alveolar macrophages inhibited LPS-induced increasing of tumor necrosis factor alpha, interleukin-6 and interleukin-1β. Luciferase assay showed STAT3 was a targeting of miRNA let-7c in alveolar macrophages.@*CONCLUSIONS@#MiRNA let-7c low expression in COPD can regulate inflammatory responses by targeting STAT3 in alveolar macrophage, which may provide a new target for COPD treatment strategies.

Anti-inflammatory role of microRNA let-7c in LPS treated alveolar macrophages by targeting STAT3

Objective: To explore the expression of microRNA (miRNA) let-7c and its function in chronic obstructive pulmonary disease (COPD) and alveolar macrophage cells. Methods: Real time PCR was performed to detect the expression of miRNA let-7c in the lung tissue of COPD patients and COPD model in mice. MiRNA let-7c was overexpressed in alveolar macrophages isolated from mice and its effect was measured by the production of pro-inflammation cytokines and the protein level of signal transducer and activator of transcription 3 (STAT3) as well as phosphorylation level of STAT3 after LPS stimulation. Luciferase assay was used to detect the binding of miRNA let-7c and 3'UTR of STAT3. Results: MiRNA let-7c expression was significantly lower in patients with COPD compared with control group, and the similar result was found in COPD mice and LPS stimulated alveolar macrophages. Overexpression of miRNA let-7c in alveolar macrophages inhibited LPS-induced increasing of tumor necrosis factor alpha, interleukin-6 and interleukin-1β. Luciferase assay showed STAT3 was a targeting of miRNA let-7c in alveolar macrophages. Conclusions: MiRNA let-7c low expression in COPD can regulate inflammatory responses by targeting STAT3 in alveolar macrophage, which may provide a new target for COPD treatment strategies.

Effects of keratinocytes/fibroblasts co-culture under pressure on cell proliferation and collagen synthesis / 医用生物力学

Objective To investigate effects of 3D co-culture of human keratinocytes (HKC) and human fibroblasts (HFB) under pressure on cell proliferation and collagen synthesis. Methods The HKC and HFB were planted on chitosan-gelatin scaffolds, respectively, for 2 d. The HKC-chitosan-gelatin complex (3D HKC) was cultured at air-liquid interface for 1 d to induce differentiation, and then co-cultured with the HFB-chitosan-gelatin complex (3D HFB) for 12 h. 3.4 kPa pressure was applied on the co-culture group for 24 h. The group of single culture with pressure, the group of single culture without pressure and the group of co-culture without pressure were used as control. HE staining was used to observe distribution and growth of HKC and HFB on chitosan-gelatin scaffolds. MTT method was used to test proliferation of HKC and HFB. Hydroxyproline kit was used to observe collagen concentration of the supernatant fluids. Results HE staining showed that HKC and HFB could grow confluently on chitosan-gelatin scaffolds；3.4 kPa pressure or co-culture both could promote the HKC proliferation and collagen synthesis, while restrain the HFB proliferation and collagen synthesis. Conclusions Pressure and co-culture play an important role in HKC and HFB proliferation and collagen synthesis. This research finding provides some reference for exploring the therapeutic mechanism of hyperplastic scar from clinical operation of resecting scar by transplanting tissue-engineered skin to the wound and then combined with pressure treatment.

Clinical studies of dynamic changes on the renal injury indicators of acute paraquat poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the clinical significance of dynamic changes of blood urea nitrogen (BUN), serum creatinine (Cr), serum cystatin (Cys C) and urinary protein on renal injury with paraquat poisoning.</p><p><b>METHODS</b>According to the clinical manifestation and curative effect, the clinical information was analyzed retrospectively in 35 cases of acute paraquat poisoning, survival after eight weeks as the standard. Poisoning patients were taken a fasting blood 5 ml and the middle of urinary on the 1st day, 3rd day, 7th day, 14th day, 21st day and 8 weeks after the poisoning. Then the levels of serum BUN, Cr, Cys C and urinary protein were detected by automatic biochemistry analyzer. 30 cases healthy subjects were randomly selected as normal control group, and discharged kidney disease and other diseases of urinary system history. The same laboratory subjects have been done.</p><p><b>RESULTS</b>The level of serum Bun, Cr, Cys C of survival group increased significantly compared with control group within 21 days (P < 0.05). The level of serum BUN, Cr Cys C decreased on the 14th day. The decreased level of serum Cys C was lower than that of serum BUN and Cr. The renal function of 29 cases among 35 cases survival patients recovered on 21st day. The renal function of 31 cases among 35 cases survival patients recovered 8 weeks late. The positive rate of urinary protein of survival patients was high in the early intoxication (76.9%).</p><p><b>CONCLUSION</b>Serum Cys C is sensitive indicator to reveal the kidney injury on paraquat poisoning patients and have higher value of clinical applications in the diagnosis of the kidney injury of paraquat poisoning, which sensitivity is higher than serum BUN and Cr. The kidney injury caused by paraquat poisoning is reversible.</p>

Efficacy and safety of induction therapy with alemtuzumab in kidney transplantation: a meta-analysis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Alemtuzumab, a humanized CD52 monoclonal antibody, with its profound lymphocyte depletion property, was expected to be a promising induction therapy agent for kidney transplantation (KTx). However, currently no consensus is available about its efficacy and safety. The aim of this meta-analysis was to make a profound review and an objective appraisal of this issue.</p><p><b>METHODS</b>Relevant papers were searched, essentially in the PubMed database and the Cochrane library. After a thorough review, randomized controlled trials (RCTs) comparing the outcome of KTx using alemtuzumab induction therapy (test group) with a control group were collected according to the inclusion criteria. Data of general characteristic of studies and major outcomes of Ktx were extracted and meta-analyses were performed with RevMan 4.2 software. The odds ratio (OR) with a 95% confidence intervals (CI) was the principle measurement of effect.</p><p><b>RESULTS</b>Five RCTs were included. The chi square test showed no significant between-study heterogeneity, thus fixed effect model was employed. Sub-group analysis with studies including alemtuzumab induction followed by a tacrolimus-based immunosuppressive regimen showed that the acute rejection rate (ARR) was lower relative to the control (OR = 0.59, 95% CI 0.34 - 1.01, P = 0.05). However, meta-analysis with all included studies revealed that neither ARR nor patient/graft survival rates differ significantly between the test and the control group, but the cytomegalovirus (CMV) infection rate was higher in the test group (OR 2.50, 95% CI 1.22 - 5.12, P = 0.01). A great number of the test group recipients safely remained on a regimen that was steroid-free and with a reduced dose of conventional immunosuppressive drugs.</p><p><b>CONCLUSIONS</b>Alemtuzumab induction therapy for KTx was an effective and safe protocol in the tested follow-up period. Steroid avoidance and a dose reduction of conventional immunosuppressive drugs after alemtuzumab induction therapy may have clinical importance. However, high quality RCTs with larger population and longer follow-up are needed for a more accurate and objective appraisal of this novel protocol.</p>

3-dimensional finite element analysis of periodontal stress distribution when impacted teeth are tracted / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To analyze stress around the impacted tooth by constituting a 3-dimensional finite element model of impacted tooth, consequently offer reference basis for clinic traction treatment.</p><p><b>METHODS</b>The 3-dimensional finite element model of the impacted tooth was constituted by CT scan, append pericementum and alveolar bone model was used to constitute impacted model. 3 forces were loaded to 3-dimensional finite element model and the periodontal stress of impacted tooth was calculated.</p><p><b>RESULTS</b>When force 1 was loaded to the model, the maximum stress was smaller, but the stress distribution was more average. When force 3 was loaded to the model, the maximum stress was larger, but the stress concentrated at the side of the force. When force 2 was loaded to the model, the stress distribution was medium.</p><p><b>CONCLUSION</b>When the direction of the force is in line with the central axis, the maximum stress is smaller, and the stress distribution is more average, while this has advantage to the eruption of the impacted tooth. When the direction of the force has angle with the central axis of the impacted tooth, the angle is larger, the maximum stress is larger and the stress distribution is more concentrate, and this goes against the eruption of the impacted tooth. The angle between the orientation of the traction and central axis of the impacted tooth is smaller, there are more advantages to the eruption of the impacted tooth. So the angle should be properly selected in order to make sure of the eruption of the impacted tooth. When the angle is quite large, more anchorage is needed to resist to the large force.</p>

Correlation between expression of matrix metalloproteinase-2 and angiogenesis in esophageal carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study expression of MMP-2 in relation to microvessel density (MVD) in esophageal carcinoma.</p><p><b>METHODS</b>Forty-eight specimens of esophageal carcinoma (Ec) and 17 specimens of grade II + III epithelial dysplasia (Dy) close to the tumor and 12 specimens of normal tissue (Nt) along the incisional margin were examined by S-P immunohistochemical staining with anti-MMP-2 monoclonal antibody. An anti-CD34 monoclonal antibody was used to show MVD.</p><p><b>RESULTS</b>MMP-2 expression in Ec was remarkably higher than that in Dy, which was higher than that in Nt. MMP-2 expression in Ec and Dy was significantly correlated with MVD in the tumor and nearby tissue. MMP-2 expression and MVD in Ec significantly associated with lymph node metastasis.</p><p><b>CONCLUSION</b>Expression of MMP-2 plays an important role in angiogenesis and lymph node metastasis of esophageal cancer.</p>

Function of a novel brain-specific gene LRRC4 / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the suppressive effect of LRRC4 gene on human glioma U251 cells and further investigate its biological functions.</p><p><b>METHODS</b>H&E, DNA and AgNORs stainings were performed on LRRC4-transfected U251 cells, mock-transfected U251 cells and non-transfected U251 cells, respectively. Quantitative analysis including cell morphometry, DNA content, DNA ploidy, silver stained argyrophilic nucleolar organizer regions (AgNORs) were investigated by image analysis. Flow cytometry was employed to determine the difference of cell cycle distribution and MTT staining was used to elucidate the activity of the LRRC4-transfected U251 cells.</p><p><b>RESULTS</b>The morphological cell parameters such as area, perimeter and diameter, DNA content, chromosomal aneupoloidy, mean area of AgNORs particles and mean nucleus area of the LRRC4-transfected U251 cells were remarkably decreased compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). Meanwhile, significant accumulation of cells in G(0)/G(1) phase but decrease of cells in S and G(2)/M phase, was observed in transfected U251 cells compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). MTT staining showed that proliferation activity of both the mock- and non-trasfected U251 cells was significantly higher than that of the U251 cells transfected with LRRC4 gene (P < 0.01).</p><p><b>CONCLUSION</b>LRRC4 gene might be involved in tumor suppression by restraining DNA synthesis and the nucleoli organizer regions-associated proteins, keeping the cell cycles in phase G(0)/G(1) and reducing proliferation activity of the glioma cells. Morphometry combined with other techniques such as flow cytometry and MTT staining can well elucidate the biological function of novel genes.</p>