Abstract:Objective：This study aims to investigate the effect of bardoxolone methyl （CDDO-Me） on the hypoxia-induced activation of human pulmonary artery adventitia fibroblasts（PAAF）and potential mechanism. Methods：Human PAAF were cultured in vitro and randomly divided into four groups：the normoxia group（21% oxygen），the hypoxia group（1% oxygen），the hypoxia plus CDDO-Me group and the normoxia plus CDDO-Me group. Cell viability was determined by CCK-8 assay. Transwell assay was carried out to assess cell migration. The levels of reactive oxygen species（ROS），malondialdehyde（MDA），glutathione（GSH）and superoxide dismutase（SOD）were detected to evaluate the level of oxidative stress. The levels of transforming growth factor（TGF）-β1，tumor necrosis factor（TNF）-α and interleukin（IL）-1β were detected by ELISA assay. The expression of α-smooth muscle actin （α-SMA） and nuclear translocation of NF-κB（p65） were detected by immunofluorescence assay. The protein levels of α-SMA，collagen Ⅰ，vimentin，phospho-Smad3 and Smad3，phospho-p65 and p65 were determined by Western blot. Results：CDDO-Me treatment（62.5，125.0，250.0，500.0 nmol/L）decreased hypoxia-induced elevations of cell viability in a concentration dependent manner，which showed significant inhibition at concentration of 250.0 nmol/L and 500.0 nmol/L. CDDO-Me（500.0 nmol/L）remarkably inhibited hypoxia-induced migration and myofibroblast transformation of PAAF，which reflected in improvement of hypertrophy，decreases in expressions of α-SMA，collagenI and vimentin. In addition，CDDO-Me（500.0 nmol/L）significantly inhibited hypoxia-induced increased levels of ROS and MDA，decreased levels of GSH and SOD，and improved the ability of antioxidation. Hypoxia increased the secretion of TGF-β1 and activate TGF-β1/Smad3 signaling pathway in PAAF，which was attenuated by CDDO-Me（500.0 nmol/L）. Besides，hypoxia-induced nuclear translocation of p65，phosphorylation of p65 protein，and the secretion of TNFα and IL-1β were inhibited by CDDO-Me treatment. Conclusion：CDDO-Me can inhibit hypoxia-induced proliferation，migration，myofibroblast transformation of PAAF，improve the antioxidation ability，and inhibit TGF-β1/Smad3 and NF-κB signaling pathway in PAAF.

Abstract:Objective：This study aims to investigate the effects of fasudil dichloroacetate（FDCA）on hypoxic pulmonary hypertension（HPH） in rats and the potential mechanisms. Methods：SD rats were randomly divided into 4 groups：control（CON） group，CON+FDCA group，chronic hypoxia（CH）group，and CH+FDCA group. The rats in CH and CH+FDCA groups were exposed to chronic hypoxia with（10.0 ± 0.1）% oxygen for 28 days，and the rats in CON+FDCA and CH+FDCA groups were given FDCA［43.3 mg/（kg·d）］intragastrically from the 15th day of hypoxia. After 28 days of hypoxia，right ventricular systolic pressure（RVSP），and right ventricular hypertrophy index（RVHI）were assessed. The morphological changes of pulmonary vessels and right ventricle were evaluated by hematoxylin-eosin staining，α-SMA immunohistochemical staining and Masson’s trichome staining. ELISA was used to analyze the contents of interleukin（IL）-1β，IL-6，tumor necrosis factor-α（TNF-α），the levels of myosin light chain kinase（MLCK） and myosin light chain phosphorylase（MLCP） in lung. The expression of ROCK1 and ROCK2 in rat lung tissues was measured by Western blot assay. Results：FDCA alleviated hypoxia-induced increments of RVSP and RVHI，as well as cardiomyocytes hypertrophy. In addition，FDCA suppressed hypoxia-induced pulmonary artery wall hypertrophy，pulmonary vessel muscularization and perivascular fibrosis. ELISA results showed that FDCA inhibited hypoxia-induced upregulation of IL-1β，IL-6 and TNF-α，and restored the expression of MLCK and MLCP in lung. Moreover，FDCA reversed hypoxia-induced upregulation of ROCK1 and ROCK2 in the lung tissues of HPH rats. Conclusion：FDCA alleviates HPH by inhibiting chronic hypoxia-induced pulmonary vascular contraction and remodeling，as well as suppressing inflammation in lung. These results indicate that FDCA could be a potential compound for treating HPH.

Abstract:Objective：The aim of the present study was to investigate the protective effects and mechanisms of of vitamin D3（Vit D3）on lipopolysaccharide（LPS）-induced liver injury in mice. Methods：Forty C57BL/6 mice were randomly assigned to 4 groups（n=10）as follows：control group，vitamin D3 group（Vit D3），model group（LPS），and treatment group（LPS+Vit D3）. Acute liver injury of mice in model group and treatment group was induced by the intraperitoneal injection of 15 mg/kg LPS. Mice in Vit D3 group and treatment group were given 2.5 μg/kg vitamin D3 at the time points of 0 h，8 h，16 h after LPS injection，while mice in the control and model groups were treated with an equivalent volume of 0.9% sodium chloride solution. After 24 h，all mice were anesthetized with chloral hydrate. Blood and livers of mice were collected for subsequent experiments. Results：Vitamin D3 decreased the levels of alanine aminotransferase（ALT）and aspartate aminotransferase（AST）in serum，increased the activity of antioxidant enzymes in the liver tissues compared with those in the model group and attenuated liver pathologic changes. In addition，vitamin D3 downregulated the serum levels of tumor necrosis factor-α（TNF-α），interleukin -1β（IL-1β） compared with those in the model group. Meanwhile，vitamin D3 downregulated the protein expression of IL-1β，TNF-α，NF-κB p65 and NF-κB p50，upregulated the expression of nuclear factor-erythroid 2p45-related factor 2（Nrf2），heme oxygenase-1（HO-1） and vitamin D receptor（VDR） in liver tissues compared with those in the model group. Conclusion：Vitamin D3 showed a protective effect against LPS-induced acute liver injury in mice，which may be partly due to the inhibition of oxidative stress and inflammation via the VDR pathway.

Abstract:Objective：This study aims to explore the role of M1-polarized macrophages in the pathogenesis of endothelial-to-mesenchymal transition（EndMT）and chronic allograft dysfunction（CAD）. Methods：The GSE21374 transcriptome array from GEO public database was downloaded，and cibersort software was used to analyze the immune cell infiltration status in allograft tissues of CAD patients. Then the allograft tissues were collected from patients diagnosed with CAD in our center. Immunofluorescence，polymerase chain reaction（PCR）and Western blot（WB）were used to observe infiltration of M1-polarized macrophages in the renal allografts. Finally，in vitro lipopolysaccharide（LPS）and interferon-γ（IFN-γ） were used to induce the polarization of Raw264.7 macrophage cell line into M1 macrophages. A transwell chamber was used to establish a co-culture system for the M1 macrophages and Balb/c mouse-derived aortic endothelial cells. PCR and WB assays，as well as cell immunofluorescence，were performed to examine the expression of CD31 and α-SMA in the mouse-derived aortic endothelial cells. Results：Based on the public database，monocytes and macrophages were observed to be highly expressed in allograft tissues from CAD patients（P < 0.05）. Similarly，significant infiltration of CD68（+）iNOS（+）M1 macrophages in glomerulus and interstitial area of allograft with CAD was observed，and PCR results showed that iNOS（inducible nitric oxide synthase），the surface marker of M1 macrophages was increased significantly compared to the normal kidney tissues（P < 0.05）. Co-cultured with M1 macrophages，the EndMT of endothelial cells was promoted. Conclusion：We have observed significant M1 macrophages infiltration in the allograft tissues diagnosed with CAD，which may induce the occurrence of EndMT and CAD progression in renal transplant recipients.

Abstract:Objective：This study aims to investigate the effect of the Pseudomonas aeruginosa secreted protein Pec1 on phagocytosis of MH-S cells. Methods：The recombinant protein Pec1 was prepared by PCR amplification，plasmid construction，induced expression and protein purification. The effect of the recombinant protein Pec1 on the proliferation of MH-S cells was measured by CCK-8 kit. Neutral red was used to detect the phagocytic function of MH-S cells，and the phagocytosis of heat- inactivated Pseudomonas aeruginosa was observed by fluorescence microscope. Results：The recombinant protein Pec1 with the molecular weight of 30.5 kDa was successfully expressed and purified. Pec1 had a concentration-dependent inhibitory effect on the proliferation of MH-S cells. Pec1 decreased the endocytosis of neutral red and inhibited the phagocytosis of heat-inactivated Pseudomonas aeruginosa by MH-S cells. Conclusion：The Pseudomonas aeruginosa secreted protein Pec1 can inhibit the phagocytosis of MH-S cells.

Abstract:Objective：This study aims to explore the effect of c-FLIP（L）in pulmonary fibrosis and its pathological mechanism. Methods：Based on bleomycin（BLM）induced idiopathic pulmonary fibrosis in mice，the expressions of c-FLIP and E-cadherin were analyzed by IHC staining. Further investigations in A549 cells overexpressing c-FLIP（L），the expression levels of E-cadherin，N-cadherin and Vimentin mRNA were examined by qRT-PCR，and Smad activation induced by transforming growth factor （TGF）-β1 was detected by luciferase reporter assay and Western blot. Results：The increased c-FLIP expression was observed and associated with a decrease of E-cadherin expression. C-FLIP（L） overexpression resulted in the changes on E-cadherin，N-cadherin and Vimentin expressions in A549 cells. Furthemore，overexpression of c-FLIP（L） enhanced TGF-β-induced Smad activation，and knocking down c-FLIP（L） blocked the TGF-β1-induced EMT progress. Conclusion：C-FLIP（L） may be a promoting factor in the development of fibrosis via regulating EMT progress.

Abstract:Objective：This study aims to study the inhibitory effect of α7 nicotinic acetylcholine receptor（α7-nAChR）on chondrocyte apoptosis，providing a new idea and research strategy for the clinical treatment and study of osteoarthritis（OA）. Methods：Mice were randomly divided into the following groups：control group，monosodium iodoacetate（MIA） treatment alone group，MIA+Nic（Nicotine 0.5 mg/kg or 1 mg/kg）treatment group and MIA+Nic+MLA（methyllycaconitine）treatment group. A mouse model of OA induced by injection of MIA was used to study the effects of nicotine on joint pain，cartilage degeneration and chondrocyte apoptosis. Mechanical withdrawal sensitivity was detected using Von Frey hairs at 7，14，and 21 days after MIA injection. Cartilage degeneration was assessed using cartilage degeneration score and aggrecan loss score at 21 days after injection. Apoptosis of articular cartilage was determined by terminal deoxynucleotidyl transferase dUTP Nick end labelling（TUNEL） staining. Meanwhile，Western blot was used to detect the expression of apoptosis-related proteins Bcl-2，Bax，cleaved caspas-9 and caspase-9. Results：After model preparation of 21 days，the latency threshold of mechanical foot retraction reflex in MIA group decreased significantly to（0.28 ± 0.02）g，the scores of articular cartilage degeneration and proteoglycan loss increased to （5.33 ± 1.19） and （2.33 ± 0.27），respectively，and the apoptosis rate of articular chondrocytes increased to（31.83 ± 3.89）%. Nic（1.0 mg/kg）treatment reduced pain behavior（P < 0.05），cartilage degeneration（P < 0.05） and chondrocyte apoptosis induced by MIA（P < 0.05）. In addition，nicotine treatment reduced MIA-induced down-regulation of Bcl-2，up-regulation of Bax and cleaved caspase-9/caspase-9 ratio levels（P < 0.05）. The benefit of nicotine was abolished by a selective α7 nicotinic receptor blocker MLA in vivo. Conclusion：The activation of α7-nAChR has a protective effect on cartilage damage in OA model mice and exerts an anti-chondrocyte apoptosis effect by inhibiting the mitochondrial apoptotic pathway.

Abstract:Objective：This study aims to determine the function of WNT7b in the progression of malignant melanoma and the underlying mechanism by which it promotes the migration and invasion of melanoma cells. Methods：Intrinsic WNT7b expression level was examined in A875，SK-MEL-28，and A375 cell lines by qRT-PCR and Western blot. The intervention of WNT7b in A375 cell line was performed by transiently transfected with small interfering RNA，and its effect on the cellular function was also analyzed by flow cytometry analysis，wound healing and Transwell assay，respectively. Then Western blot was used to detect the expression of epithelial-mesenchymal transition（EMT）related markers and members of matrix metalloproteinase（MMP）family. Results：WNT7b was expressed in A375，SK-MEL-28 and A875 cell lines，among which A375 presents the highest WNT7b expression. Our cellular study showed that upon knockdown of WNT7b expression in human A375 cell line，the cell cycle was arrested at G1 phase，the cell migration and invasion ability were significantly inhibited. In addition，WNT7b knockdown led to the up-regulation of E-cadherin and down-regulation of N-cadherin，Vimentin and Snail1. Further detection of MMP family members showed that the expression of MMP-2，MMP-7 and MMP-9 was reduced. Conclusion：WNT7b may induce EMT through MMPs in human malignant melanoma cells，thereby promoting cells migration and invasion.

Abstract:Objective：This study aims to prepare a bio-collagen coating on pure titanium surface and investigate its effect on osteoblast behaviors. Methods：The type I collagen was made into gel and adhered to the surface of pure titanium specimens to form bio-collagen coating. The smooth titanium surface was used as the control group，and the collagen coated titanium surface was used as the experimental group. The surface microtopography of specimens was observed by scanning electron microscopy（SEM）. The surface elemental compositions of specimens were analyzed via X-ray photoelectron spectroscopy（XPS）. The groups on specimen surfaces were detected with fourier transform infrared spectrometer（FT-IR），and the surface hydrophilicity of specimens was tested by water contact angle meter. The adhesion，proliferation and differentiation of osteoblasts on titanium surfaces were evaluated by confocal laser scanning microscope（CLSM），CCK-8，alkaline phosphatase（ALP） detection and Western blot after the osteoblasts（MC3T3-E1） were cultured in vitro. Results：SEM observation found the depth of mechanical scratches decreased on the surface of experimental specimens. XPS elemental analysis and FT-IR detection confirmed the presence of a collagen coating on titanium surface. Contact angle test results showed that the experimental group had better surface hydrophilicity than the control group. The results of in vitro study exhibited that collagen coated titanium surface could promote the adhesion and proliferation activity of osteoblasts and up-regulate ALP activity as well as the expressions of osteogenic-related proteins including Runx2，osterix and OCN. Conclusion：The modification of collagen gel coating on pure titanium surface can enhance its hydrophilicity and promote adhesion，proliferation and differentiation of osteoblasts.

Abstract:Objective：This study aims to explore a scoring system for predicting the risk of delayed bleeding after endoscopic submucosal dissection（ESD）for early gastric cancer（EGC）. Methods：A retrospective analysis was performed on the data of 296 patients with early gastric cancer（including high-grade intraepithelial neoplasia） who underwent ESD treatment in the Gastroenterology Department of the First Affiliated Hospital of Soochow University from February 2016 to November 2020. Multivariate logistic regression analysis was performed to investigate independent risk factors of delayed bleeding after ESD in EGC. Weighted points was proportionally assigned based on β regression coefficients value to construct the scoring system. According to the scores and the incidence of delayed bleeding，the receiver operating characteristic curve（ROC curve） was plotted，the area under the curve（AUC） was calculated，and risk stratification based on the scores was performed. Results：Multivariate analysis revealed that submucosal fibrosis（OR=12.163，95%CI：2.735-54.102），resected specimen diameter＞4 cm（OR=19.091，95%CI：3.726~97.819），use of anticoagulants and/or antiplatelet drugs were independent risk factors for delayed bleeding after ESD in EGC（all P＜0.05）. Scoring system：1 point for the use of anticoagulants and/or antiplatelet drugs，2 points for submucosal fibrosis of the lesion，and 2 points for resected specimen diameter＞4 cm. The AUC value of the scoring system was 0.85（95%CI：0.753-0.947，P＜0.001），of which the cut-off value was 1.5. According to the cut-off value，a score of 0-1 was defined as the low-risk group，and a score of 2-5 was defined as the high-risk group for EGC after ESD. The incidence of delayed bleeding after ESD in the low-risk group was 1.03%（2/194），and in the high-risk group was 16.67%（17/102）. The differences between the two groups were statistically significant（χ2=27.208，P＜0.001）. Conclusion：The predictive scoring system constructed by the three independent risk factors，which were submucosal fibrosis of the lesion，the use of anticoagulants and/or antiplatelet drugs，and resected specimen diameter＞4 cm，can effectively identify high-risk patients with delayed bleeding after ESD for EGC.

Abstract:Objective：This study aims to evaluate the safety and efficiency of the percutaneous left atrial appendage occlusion（LAAO） in nonvalvular atrial fibrillation（NVAF） patients with coronary heart disease（CAD）. Methods：Total 51 NVAF patients were consecutively accepted LAAO from June 2015 to July 2017. Patients were divided in two groups：15 with CAD and 36 without CAD. All patients were followed up after discharge with transesophageal echocardiography（TEE） examination. Results：Patients with CAD had higher incidence of hemorrhagic stroke or major bleeding than those without CAD. The incidence of end-point events had no statistical difference between CAD group and non-CAD group. Compared with CAD patients who accepted long-term antithrombotic medication，there was a further reduction of hemorrhage in CAD patients who accepted LAAO group. Moderate or severe left atrial spontaneous echo contrast was correlated with the composite end point events. Conclusion：There are similar safety and effectiveness for LAAO procedure in NVAF patients with or without CAD. Meanwhile，left atrial spontaneous echo contrast is a predictive factor of LAAO in NVAF patients combined CAD.

Abstract:Objective：Visual hallucination（VH）is the most common psychiatric symptom in Parkinson’s disease（PD），which is closely related to the impaired quality of life and adversely affects the prognosis of patients. However，its pathogenesis is not yet clear.VH has been found to have relationship with disturbance in brain structures，visual pathways and attention networks，as well as neurotransmitters，and there is strong evidence to support these findings in epidemiology，histopathology，clinical features and so on. Based on the results of multidisciplinary research at home and abroad，this paper is to discuss the pathogenesis of VH in PD.

Abstract:Breast cancer is the most common malignant tumor among women. In recent years，research data show that early diagnosis and treatment can significantly reduce the mortality of patients with cancer. Positron mission computed tomography（PET/CT）is a noninvasive and systemic imaging method which integrates function and anatomy，and plays an important role in the clinical staging and guiding treatment of brenst cancer. 18F-fluorodeoxyglucose（18F-FDG） is the most widely used imaging agent. Because of non-specificity，its application in breast cancer patients is limited to some extent. Therefore，for the past few years，there were more researches on the specific receptor imaging agents for breast cancer patients. This paper reviews the development and clinical application of new molecular targeted positron drugs in PET imaging of breast cancer.

Abstract:The rapid development of pediatric intensive care medicine has significantly improved the success treatment of critically ill children in recent years. Although some children could survive，they are prone to secondary problems such as decreased motor ability and cognitive dysfunction，which seriously affect the quality of life of these children. A large number of studies have confirmed the feasibility and effectiveness of early rehabilitation treatment for critically ill children. In this article，we reviewed the objects，timing of intervention，specific rehabilitation techniques，and rehabilitation evaluation of early children rehabilitation in intensive care unit，aiming to provide clinical treatment for early rehabilitation treatment in pediatric intensive care unit.