Reparação da parede abdominal com membranas acelulares de pericárdio bovino - Parte I - Achados clínicos e radiológicos / Abdominal wall repair with acellular bovine pericardial membranes - Part I - Clinical and radiological findings

Introdução: Reduzindo os índices de recidiva de forma impactante, o emprego de biomateriais como "telas de reforço" na reparação de diferentes defeitos da parede abdominal tornou-se rotina quase obrigatória para o sucesso dessas reparações. A partir da década de 1990 houve a introdução de matrizes biológicas acelulares, iniciando-se assim uma nova era na reparação dos defeitos da parede abdominal. O objetivo é avaliar a funcionalidade do pericárdio bovino acelularizado em reparações da parede abdominal. Método: Trinta pacientes foram submetidos a reparação de defeitos da parede abdominal, com biopróteses acelulares de pericárdio bovino, perfazendo um total de 40 implantes anatomicamente individualizados. O seguimento médio foi de 31 meses, sendo os pacientes avaliados clinicamente e radiologicamente. Em três casos foram feitas biópsias das áreas implantadas permitindo análise histológica do material. Resultados: Não se observou recidiva das herniações em nenhum dos casos, tanto clinica como radiologicamente. Também não houve registro de hematomas, infecções ou qualquer fenômeno de natureza reacional local ou sistêmica. Radiologicamente, não foi possível visualizar as matrizes no local de implantação em qualquer dos períodos de pós-operatório analisados. Conclusão: As matrizes mostraram similaridade às demais membranas biológicas descritas na literatura internacional. Representando uma importante atualização e evolução conceitual, as membranas acelulares de pericárdio bovino podem ser incorporadas ao arsenal terapêutico nas reparações de parede abdominal.

Introduction: Reducing recurrence rates significantly, the use of biomaterials as "reinforcement meshes" in the repair of different abdominal wall defects has become an almost mandatory routine for the success of these repairs. From the 1990s onwards, acellular biological matrices were introduced, thus beginning a new era in the repair of abdominal wall defects. The objective is to evaluate the functionality of the acellularized bovine pericardium in abdominal wall repairs. Method: Thirty patients underwent repair of abdominal wall defects using acellular bovine pericardium bioprostheses, making a total of 40 anatomically individualized implants. The average follow-up was 31 months, with patients being evaluated clinically and radiologically. In three cases, biopsies were taken from the implanted areas, allowing histological analysis of the material. Results: No recurrence of herniations was observed in any of the cases, both clinically and radiologically. There were also no records of bruises, infections or any phenomenon of a local or systemic reaction nature. Radiologically, it was not possible to visualize the matrices at the implantation site in any of the postoperative periods analyzed. Conclusion: The matrices showed similarity to other biological membranes described in the international literature. Representing an important update and conceptual evolution, acellular bovine pericardial membranes can be incorporated into the therapeutic arsenal in abdominal wall repairs.

Reparação da parede abdominal com membranas acelulares de pericárdio bovino - Parte II - Análises histológicas e morfométricas / Repair of the abdominal wall with acellular bovine pericardial membranes - Part II - Histological and morphometric analyses

Introdução: Análise histológica é a principal ferramenta de avaliação de biopróteses acelulares, em sua maioria em caráter experimental. O objetivo é analisar histologicamente a matriz acelular de pericárdio bovino em reparações de parede abdominal implantada em humanos. Método: De uma série de 30 reparações com a membrana, 3 pacientes foram submetidas a revisão cirúrgica não relacionada aos implantes, aos 13, 22 e 23 meses de pós-operatório, obtendo-se biópsias das áreas previamente implantadas. Além da avaliação dos aspectos básicos de biocompatibilidade e neoformação tecidual, as lâminas foram digitalizadas e submetidas a análise computadorizada com o software ImageJ para quantificação da cinética de degradação das membranas, associada à análise da dimensão fractal das amostras. Os valores obtidos para porcentagens de membrana residual tiveram suas médias comparadas por análise de variância (ANOVA) e pelo teste T de Student não pareado, também utilizado para os valores da quantificação da dimensão fractal. Resultados: Foi demonstrada a biocompatibilidade do material, com neoformação tecidual, deposição de colágeno e tecido celularizado de aspecto normal, sem reações locais importantes. Fragmentos residuais da membrana foram quantificados em 40%±7% aos 13 meses, em 20%±6% aos 22 meses e em 17%±6% aos 23 meses de pós-operatório, com a análise da dimensão fractal indicando uma progressiva degradação dos implantes, com significância estatística entre 13 meses e as amostras tardias. Conclusão: Os resultados atestaram a funcionalidade do pericárdio bovino acelular sob diferentes níveis de estresse mecânico nas reparações da parede abdominal em humanos.

Introduction: Histological analysis is the main tool for evaluating acellular bioprostheses, mostly on an experimental basis. The objective is to histologically analyze the acellular matrix of bovine pericardium in abdominal wall repairs implanted in humans. Method: From a series of 30 repairs with the membrane, 3 patients underwent surgical revision unrelated to the implants at 13, 22, and 23 months postoperatively, obtaining biopsies of the previously implanted areas. In addition to evaluating the basic aspects of biocompatibility and tissue neoformation, the slides were digitalized and subjected to computerized analysis with the ImageJ software to quantify the kinetics of membrane degradation associated with the analysis of the fractal dimension of the samples. The values obtained for percentages of residual membrane had their means compared by analysis of variance (ANOVA) and the unpaired Student's T test, also used for the fractal dimension quantification values. Results: The biocompatibility of the material was demonstrated, with tissue neoformation, collagen deposition, and cellularized tissue with a normal appearance without important local reactions. Residual fragments of the membrane were quantified at 40%±7% at 13 months, at 20%±6% at 22 months, and at 17%±6% at 23 months postoperatively, with the analysis of the fractal dimension indicating a progressive degradation of implants, with statistical significance between 13 months and late samples. Conclusion: The results confirmed the functionality of the acellular bovine pericardium under different levels of mechanical stress in abdominal wall repairs in humans.

Effect of Guipitang on ERK1/2 and p38 MAPK in Rats with Myocardial Ischemia / 中国实验方剂学杂志

ObjectiveTo explore the therapeutic effect and mechanism of Guipitang on rats with myocardial ischemia. MethodFifty SD rats were divided into five groups： a control group， a model group， low and high-dose Guipitang （7.52， 15.04 g·kg-1） groups， and a trimetazidine group （0.002 g·kg-1）. By intragastric administration of vitamin D3 and feeding rats with high-fat forage and injecting isoproterenol， the rat model of myocardial ischemia was established. After drug treatment of 15 d， an electrocardiogram （ECG） was performed to analyze the degree of myocardial injury. A fully automatic biochemical analyzer was used to detect the changes in the serum levels of total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）. Hematoxylin-eosin （HE） staining and Masson staining were used to observe myocardial histopathological changes. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect cardiomyocyte apoptosis. Western blot was adopted to detect the protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， phospho-ERK1/2 （p-ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， phospho-p38 MAPK （p-p38 MAPK）， B-cell lymphoma-2 （Bcl-2）-associated X （Bax）， Bcl-2， and cleaved cysteine aspartate proteolytic enzyme （cleaved Caspase-3）. ResultCompared with the control group， the ECG S-T segment decreased in the model group. The serum levels of TC， TG， and LDL-C were increased significantly （P<0.05）. The arrangement of myocardial tissue was disordered， and the proportion of cardiomyocyte apoptosis increased. The protein levels of cleaved Caspase-3， Bax， and p-p38 MAPK in the heart were increased， and the Bcl-2 expression was decreased （P<0.05）. Compared with the model group， the S-T segment downward shift was restored in the low and high-dose Guipitang groups and trimetazidine group， and the levels of TC， TG， and LDL-C were decreased. The protein expression of cleaved Caspase-3 and Bax in the heart dropped， and p-p38 MAPK and p-ERK1/2 protein expressions increased significantly （P<0.05）. The degree of myocardial injury was alleviated， and the proportion of cardiomyocyte apoptosis decreased. Bcl-2 protein expression was increased significantly in the low-dose Guipitang group （P<0.05）. ERK1/2 and p38 MAPK proteins had no significant difference among different groups. ConclusionGuipitang could alleviate myocardial injury and inhibit cardiomyocyte apoptosis in rats by activating the expression of ERK1/2 and p38 MAPK.

Lactadherin immunoblockade in small extracellular vesicles inhibits sEV-mediated increase of pro-metastatic capacities

BACKGROUND: Tumor-derived small extracellular vesicles (sEVs) can promote tumorigenic and metastatic capacities in less aggressive recipient cells mainly through the biomolecules in their cargo. However, despite recent advances, the specific molecules orchestrating these changes are not completely defined. Lactadherin is a secreted 0protein typically found in the milk fat globule membrane. Its overexpression has been associated with increased tumorigenesis and metastasis in breast cancer (BC) and other tumors. However, neither its presence in sEVs secreted by BC cells, nor its role in sEV-mediated intercellular communication have been described. The present study focused on the role of lactadherin-containing sEVs from metastatic MDA-MB-231 triple-negative BC (TNBC) cells (sEV-MDA231) in the promotion of pro-metastatic capacities in non-tumorigenic and non-metastatic recipient cells in vitro, as well as their pro-metastatic role in a murine model of peritoneal carcinomatosis. RESULTS: We show that lactadherin is present in sEVs secreted by BC cells and it is higher in sEV-MDA231 compared with the other BC cell-secreted sEVs measured through ELISA. Incubation of non-metastatic recipient cells with sEV- MDA231 increases their migration and, to some extent, their tumoroid formation capacity but not their anchorage-independent growth. Remarkably, lactadherin blockade in sEV-MDA231 results in a significant decrease of those sEV-mediated changes in vitro. Similarly, intraperitoneally treatment of mice with MDA-MB-231 BC cells and sEV-MDA231 greatly increase the formation of malignant ascites and tumor micronodules, effects that were significantly inhibited when lactadherin was previously blocked in those sEV-MDA231. CONCLUSIONS: As to our knowledge, our study provides the first evidence on the role of lactadherin in metastatic BC cell-secreted sEVs as promoter of: (i) metastatic capacities in less aggressive recipient cells, and ii) the formation of malignant ascites and metastatic tumor nodules. These results increase our understanding on the role of lactadherin in sEVs as promoter of metastatic capacities which can be used as a therapeutic option for BC and other malignancies.

ENAM gene polymorphisms associated with dental anomalies in individuals with cleft lip and palate

Aim: This study aimed to investigate the occurrence of enamelin gene (ENAM) single nucleotide polymorphisms (SNP) and ENAM polymorphism association with dental anomalies (DA) in individuals with unilateral or bilateral cleft lip and palate (CLP). Methods: Saliva samples were collected from 147 individuals aged between 6 and 15 years-old, both genders, and divided into 4 groups: Group 1 (G1) - CLP and DA; Group 2 (G2) - CLP without DA; Group 3 (G3) - without CLP with DA; Group 4 (G4) - without CLP and DA. The genomic DNA was extracted from saliva samples and the following ENAM SNPs markers were genotyped: rs3796703, rs3796704, rs3796705, rs7671281, rs2609428, and rs35951442. Fisher exact and Pearson's Chi-square tests statistically analyzed the results (α=5%). Results: Individuals without CLP with DA (Group 3 - 19.2%) showed statistically higher prevalence of SNP rs2609428 heterozygotes (p=0.006) than individuals with CLP and DA (Group 1 - 0%). Individuals without CLP (10%) exhibited statistically higher prevalence of mutated heterozygotes/homozygous (p=0.028) than in individuals with CLP (1.3%). Conclusion: SNP rs2609428 marker of ENAM gene may be associated with dental anomalies in individuals without cleft lip and palate

Rol de trampas extracelulares de neutrófilos en el pronóstico de las infecciones del sistema respiratorio / Role of neutrophil extracellular traps in the prognosis of respiratory tract infections

Las trampas extracelulares de neutrófilos (NET, por sus siglas en inglés) han surgido recientemente como un vínculo potencial entre la inmunidad y la inflamación, que podría cumplir un papel clave en la patogénesis de las infecciones de vías respiratorias. El objetivo de esta revisión es determinar su rol como marcador pronóstico en enfermedades infecciosas de vías respiratorias. Para la elaboración de este artículo de revisión narrativa se consultaron las publicaciones disponibles a través de una búsqueda automatizada en las bases de datos de PubMed, Scopus y Embase. Las concentraciones elevadas de trampas extracelulares de neutrófilos (cfADN, complejos de mieloperoxidasas-ADN) en pacientes con cuadro clínico grave por infecciones de vías respiratorias, se relacionan con una estancia hospitalaria más larga, periodo prolongado de administración de antibióticos, aumento del riesgo de ingreso a la UCI, necesidad de ventilación mecánica, disfunción orgánica e incluso la muerte (p ≤ 0,05). A pesar de no contar con un parámetro de medición estandarizado, el exceso de trampas extracelulares de neutrófilos se corresponde con la gravedad del daño tisular observado en pacientes con infecciones de vías respiratorias, esto revela el importante rol pronóstico de la respuesta de los neutrófilos y del proceso de la NETosis en las enfermedades infecciosas pulmonares

Neutrophil extracellular traps (NET) have recently emerged as a potential link between immunity and inflammation, which could play a key role in the pathogenesis of respiratory tract infections. This review aims to determine the role of neutrophil extracellular traps as prognostic markers in respiratoria tract infectious diseases. For this article a literature review was undertaken, consulting available publications through an automated search in PubMed, Scopus, and Embase databases. High concentrations of neutrophil extracellular traps (cfDNA, Myeloperoxidase-DNA complexes) in patients with severe clinical presentation due to respiratory tract infections are related to a longer length of hospital stay, prolonged period of antibiotic administration and increased risk of admission to the ICU, need for mechanical ventilation, organ dysfunction and even death (p ≤ 0.05). Despite not having a standardized measurement parameter, the excess of neutrophil extracellular traps corresponds to the severity of tissue damage observed in patients with respiratory tract infections, revealing the important prognostic role of the neutrophil response and NETosis process in pulmonary infectious diseases

Inhibition of ANGPT2 activates autophagy during hypertrophic scar formation via PI3K/AKT/mTOR pathway

Abstract Background Hypertrophic scar (HS), a fibroproliferative disorder caused by aberrant wound healing following skin injuries such as burns, lacerations and surgery, is characterized by invasive proliferation of fibroblasts and excessive extracellular matrix (ECM) accumulation. The dysregulation of autophagy is the pathological basis of HS formation. Previously, angiopoietin-2 (ANGPT2) was found to be overexpressed in HS fibroblasts (HSFs) compared with normal skin fibroblasts. However, whether ANGPT2 participates in the process of HS formation and the potential molecular mechanisms are not clear. Objective This study is intended to figure out the role of ANGPT2 and ANGPT2-mediated autophagy during the development of HS. Methods RT-qPCR was used to detect ANGPT2 expression in HS tissues and HSFs. HSFs were transfected with sh-ANGPT2 to knock down ANGPT2 expression and then treated with MHT1485, the mTOR agonist. The effects of sh-ANGPT2 or MHT1485 on the proliferation, migration, autophagy and ECM accumulation of HSFs were evaluated by CCK-8 assay, Transwell assay and western blotting. The expression of PI3K/Akt/mTOR pathway-related molecules (p-PI3K, p-Akt and p-mTOR) was assessed by western blotting. Results ANGPT2 expression was markedly upregulated in HS tissues and HSFs. ANGPT2 knockdown decreased the expression of p-PI3K, p-Akt and p-mTOR. ANGPT2 knockdown activated autophagy and inhibited the proliferation, migration, and ECM accumulation of HSFs. Additionally, the treatment of MHT1485, the mTOR agonist, on ANGPT2-downregulated HSFs, partially reversed the influence of ANGPT2 knockdown on HSFs. Study limitations The study lacks the establishment of more stable in vivo animal models of HS for investigating the effects of ANGPT2 on HS formation in experimental animals. Conclusions ANGPT2 downregulation represses growth, migration, and ECM accumulation of HSFs via autophagy activation by suppressing the PI3K/Akt/mTOR pathway. Our study provides a novel potential therapeutic target for HS.

Bioinformatics-based design of a fusion vaccine with CTLA-4 variable region to combat Brucella

Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.

Scaffolds Vasculares Descelularizados Provenientes de Vasos Sanguíneos de Placenta Bovina / Decellularized Vascular Scaffolds Derived from Bovine Placenta Blood Vessels

Resumo Fundamento As doenças associadas ao aparelho circulatório são as principais causas de morbidade e mortalidade em todo o mundo, implicando a necessidade de implantes vasculares. Assim, a produção de biomateriais vasculares tem se mostrado uma alternativa promissora às terapias utilizadas em estudos e pesquisas relacionados à fisiologia vascular. Objetivos O presente projeto visa ao desenvolvimento artificial de vasos sanguíneos pela recelularização de scaffolds vasculares derivados de vasos placentários bovinos. Métodos A superfície corioalantoide da placenta bovina foi utilizada para produzir biomateriais descelularizados. Para a recelularização, 2,5 x 104 células endoteliais foram semeadas acima de cada fragmento de vaso descelularizado durante três ou sete dias, quando a cultura foi interrompida e os fragmentos foram fixados para análise de adesão celular. Biomateriais descelularizados e recelularizados foram avaliados por histologia básica, microscopia eletrônica de varredura e imuno-histoquímica. Resultados o processo de descelularização produziu vasos que mantiveram a estrutura natural e o conteúdo de elastina, e não foram observadas células e gDNA remanescentes. Além disso, células precursoras endoteliais se ligaram ao lúmen e à superfície externa do vaso descelularizado. Conclusão nossos resultados mostram a possibilidade de usos futuros desse biomaterial na medicina cardiovascular, como, por exemplo, no desenvolvimento de vasos artificiais.

Background Diseases associated with the circulatory system are the main causes of worldwide morbidity and mortality, implying the need for vascular implants. Thus, the production of vascular biomaterials has proven to be a promising alternative to therapies used in studies and research related to vascular physiology. Objectives The present project aims to achieve the artificial development of blood vessels through the recellularization of vascular scaffolds derived from bovine placental vessels. Methods The chorioallantoic surface of the bovine placenta was used to produce decellularized biomaterials. For recellularization, 2.5 x 104 endothelial cells were seeded above each decellularized vessel fragment during three or seven days, when culture were interrupted, and the fragments were fixed for cell attachment analysis. Decellularized and recellularized biomaterials were evaluated by basic histology, scanning electron microscopy, and immunohistochemistry. Results The decellularization process produced vessels that maintained natural structure and elastin content, and no cells or gDNA remains were observed. Endothelial precursor cells were also attached to lumen and external surface of the decellularized vessel.Conclusion: Our results show a possibility of future uses of this biomaterial in cardiovascular medicine, as in the development of engineered vessels.

Progress of Research on Tumor Extracellular Matrix and Discoidin Domain Receptor 1 / 肿瘤防治研究

Tumor extracellular matrix (ECM) is the center component of tumor microenvironment (TME), ECM diversity constitutes the inherent heterogeneity of TME that contributes to tumor growth, dormancy, drug resistance, and metastasis. Discoidin domain receptor 1 is one of the ECM receptors that interact with multiple ECM ligands. It also regulates the occurrence and development of tumors. Accordingly, DDR1 plays an increasingly important role in the prevention, diagnosis, and treatment of cancer. In this review, we primarily summarize the research of ECM and its receptors with components, regulation, cell receptors, and signaling pathways in tumor progression.

Research progress in the role of mesenchymal stem cell sheets and their derivatives in bone defect repair / 中华创伤杂志

Bone defects are mostly caused by severe trauma, infection, tumor resection and congenital malformations, which adversely affect their health and quality of life. So far, the bone defects are mainly filled with autologous or allogeneic bone grafting, which has problems such as donor shortage, secondary bone injury and scarring. In recent years, the rise of bone tissue engineering has provided a new way for repair of bone defects, in which mesenchymal stem cell (MSC) sheets prepared by using the principle of tissue engineering can well solve the above problems of autologous or allogeneic bone grafting. With the development of preparation technology, new bone defect repair materials such as decellularized extracellular matrix (ECM) sheets and MSC/ECM clumps have been derived on the basis of MSC sheets. Therefore, the authors reviewed the preparation and the role of MSC sheets and their derivatives in bone defect repair, hoping to provide a reference for basic research and clinical treatment related to bone defect repair.

Research progress on the relationship between extracellular vesicles and male infertility / 中国综合临床

Infertility is gradually becoming a major problem affecting health worldwide, and male factors also play an important role in infertility. Extracellular vesicles (EVs) are ultramicro membranous vesicles released by cells during activation or apoptosis, which play an important role in cell communication. Relevant studies have shown that extracellular vesicles contain a variety of bioactive substances and participate in infertility related pathophysiological processes by influencing the content of intercellular transmission. Therefore, we reviewed the relationship between extracellular vesicles and male infertility, and expounded the occurrence and potential treatment of male infertility from another perspective.

The diagnostic value of KRAS gene mutation in appendiceal adenocarcinoma / 中国医师杂志

Objective:To analyze the mutation characteristics of Kirsten rat sarcoma virus oncogene homology (KRAS) gene in patients with appendiceal adenocarcinoma and its relationship with the activity of Ras Raf Mitogen activated protein kinase/extracellular regulated protein kinase (MAPK/ERK) signaling pathway.Methods:A total of 41 patients with appendiceal adenocarcinoma who were treated in the Lishui Central Hospital from January 2014 to January 2020 were selected as the observation group, and 50 patients with Appendicitis who were operated at the same time were randomly selected as the control group. Clinical and follow-up data were collected, and the mutation of the KRAS gene in the patient′s tissue was measured using the snapshot method. The expression of key proteins in the MAPK/ERK signaling pathway in cancer tissue was measured using Western blotting (WB) assay. We compared the clinical characteristics and prognosis of patients with KRAS mutation and non KRAS mutation appendiceal adenocarcinoma.Results:The KRAS gene mutation rate in the observation group was higher than that in the control group (41.5% vs 10.0%), and the expression levels of p-ARAF/ARAF, p-MEK1/MEK1, and p-ERK1/ERK1 proteins were also higher than those in the control group. The differences between the groups were statistically significant (all P<0.05). The protein expression levels of p-ARAF/ARAF, p-MEK1/MEK1, p-ERK1/ERK1 in KRAS mutation patients in the observation group were significantly higher than those in non KRAS mutation patients. The proportion of stage IV, positive rates of carcinoembryonic antigen (CEA), carbohydrate antigen (CA)199 and CA125 in KRAS mutation patients were higher than those in non KRAS mutation patients, and the survival time and progression free survival time were shorter than those in non KRAS mutation patients, with statistical significance (all P<0.05). Conclusions:The mutation rate of KRAS in appendix adenocarcinoma is high, and the activation of MAPK/ERK signaling pathway caused by KRAS mutation may play a role in the pathogenesis of appendix adenocarcinoma, which has the value of in-depth research.

Intervention of modified Shenling Baizhu San on peritoneal fibrosis induced by peritoneal dialysate with different sugar concentration in rats / 中华危重病急救医学

Objective:To investigate the effect of modified Shenling Baizhu San on the pathological changes and extracellular matrix (ECM) in rats with peritoneal fibrosis induced by peritoneal dialysate fluid (PDF) with different sugar concentration and its mechanism.Methods:Seventy male Sprague-Dawley (SD) rats were randomly divided into control group, different sugar content PDF model groups and corresponding traditional Chinese medicine intervention groups, with 10 rats in each group. Peritoneal fibrosis model was reproduced by intraperitoneal injection of 100 mL·kg -1·d -1 PDF containing 1.5%, 2.5% and 4.25% sugar once a day for 8 weeks. The rats in the control group were given the same amount of normal saline. The rats in the traditional Chinese medicine intervention groups were treated with gavage of 10 mL/kg of modified Shenling Baizhu San (containing 2.014 g crude drug per liter) immediately after modeling. The PDF model groups and the control group were given the same amount of normal saline by gavage. After 8 weeks, the peritoneal ultrafiltration volume of rats in each group was measured. The peritoneal tissues were collected and stained with hematoxylin-eosin (HE), and the structural changes and thickness of the parietal peritoneum were observed under a light microscope. After Masson staining, the deposition of collagen fibers was observed under a light microscope. Western blotting was used to detect the protein expressions of E-cadherin,?α-smooth muscle actin (α-SMA) and Vimentin, the main components of ECM in parietal peritoneum. The positive expressions of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and transforming growth factor-β1 (TGF-β1) were detected by immunohistochemical staining. Results:Compared with the control group, PDF with different sugar contents could induce peritoneal fibrosis in rats, and the degree of fibrosis was gradually aggravated with the increase of sugar content, which was manifested as peritoneal thickening, increased collagen fiber deposition, decreased peritoneal ultrafiltration volume, down-regulated expressions of E-cadherin and MMP-9 in peritoneal tissue, and up-regulated expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1, and the pathological changes and ECM accumulation in peritoneal tissues were more serious in 4.25% PDF model group. After the intervention of modified Shenling Baizhu San, compared with the corresponding PDF model groups, the peritoneal fibrosis of rats was improved to varying degrees, and the effect of the 4.25% PDF+traditional Chinese medicine intervention group was more significant, the parietal peritoneum was significantly thinner (μm: 101.86±16.01 vs. 140.65±10.13, P < 0.05), collagen fiber deposition was significantly reduced, peritoneal ultrafiltration volume was significantly increased (mL: -0.01±3.45 vs. -3.53±1.84, P < 0.05), the expressions of E-cadherin and MMP-9 in peritoneal tissues were significantly up-regulated [E-cadherin protein (E-cadherin/β-actin): 0.84±0.08 vs. 0.28±0.05, MMP-9 ( A value): 0.60±0.15 vs. 0.37±0.01, both P < 0.05], and the expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1 were significantly down-regulated [α-SMA protein (α-SMA/β-actin): 0.36±0.08 vs. 1.05±0.09, Vimentin protein (Vimentin/β-actin): 0.53±0.07 vs. 1.19±0.04, TIMP-1 ( A value): 0.49±0.06 vs. 0.87±0.02, TGF-β1 ( A value): 0.67±0.04 vs. 0.89±0.10, all P < 0.05]. Conclusions:The degree of peritoneal fibrosis gradually increased with the increase of PDF sugar content in rats. Modified Shenling Baizhu San can improve peritoneal fibrosis induced by PDF with different sugar contents in rats, and the mechanism is related to the changes in the expression of fibrosis markers and ECM accumulation.

Effects of NLRP3 inflammasome activation on the proliferation, migration and extracellular matrix deposition of pancreatic stellate cells / 中华胰腺病杂志

Objective:To investigate the effects of NOD-like receptor protein 3(NLRP3) inflammasome activation on the proliferation, migration and extracellular matrix desposition of activated pancreatic stellate cells(PSCs).Methods:The rat PSCs were isolated, cultured and identified, and were divided into control group or LPS group based on the pretreatment with LPS (10 μg/ml for 24 hours) or without. The expression of NLRP3 inflammasome associated molecules in PSCs culture medium was detected by ELISA. The PSCs with NLRP3 inhibition were constructed by shRNA carrying lentivirus infection and were divided into LPS+ negative control group and LPS+ lentivirus group based on whether the cells were treated with LPS and infected by lentivirus or not. The alteration in cell proliferation and migration were detected by CCK-8 kit and transwell chamber method. The expression of extracellular matrix α-SMA and collagen in PSCs was detected by immunofluorescence staining and the expression of TGF-β mRNA was analyzed by RT-qPCR.Results:The cytoplasm of PSCs which were cultured for 24 hours was rich in bright annular lipid droplets, and the cells expressed desmin. After 7 days of culture, the cell became larger in size, the lipid droplets basically disappeared, and the cells were activated and expressed α-SMA. The expression of caspase-1, IL-1β and IL-18 in the supernatant of PSCs culture medium in LPS group were significantly higher than those in control group (1.55±0.04 vs 0.65±0.03), (2.02±0.04 vs 1.05±0.05) and (1.70±0.05 vs 0.97±0.03), respectively. After inhibiting by lentivirus infection, the expression of NLRP3 in the lentivirus group (0.25±0.04) was significantly lower than that in negative control group (0.68±0.05). In control group, LPS group, LPS+ negative control group and LPS+ lentivirus group, the A490 values was 0.61±0.02, 1.15±0.06, 0.96±0.05, and 0.56±0.01, respectively; the migrating PSCs number was (64.12±4.58), (121.67±8.02), (111.67±4.67) and (69.67±8.08)/HF, respectively; the relative expression of α-SMA was 0.78±0.05, 4.12±0.04, 3.81±0.06 and 0.88±0.05, respectively; the relative expression of collagen was 0.65±0.03, 3.43±0.02, 2.67±0.02 and 0.48±0.03, respectively; and the expression of TGF-β mRNA was 0.22±0.03, 0.89±0.01, 0.86±0.03 and 0.43±0.02, respectively. The A490 value, the migrating cells number, the expression of α-SMA, collagen and the expression of TGF-β mRNA in LPS group and LPS+ negative control group was significantly higher than those in control group and LPS+ lentivirus group, and all the differences were statistically significant (all P value <0.05). Conclusions:NLRP3 inflammasome activation may accelerate the extracellular matrix deposition and pancreatic fibrogenesis by promoting PSCs proliferation and migration ability via regulating the biological functions.

Recent progress in aptamer-based microfluidics for the detection of circulating tumor cells and extracellular vesicles / 药物分析学报

Liquid biopsy is a technology that exhibits potential to detect cancer early,monitor therapies,and predict cancer prognosis due to its unique characteristics,including noninvasive sampling and real-time analysis.Circulating tumor cells(CTCs)and extracellular vesicles(EVs)are two important components of circu-lating targets,carrying substantial disease-related molecular information and playing a key role in liquid biopsy.Aptamers are single-stranded oligonucleotides with superior affinity and specificity,and they can bind to targets by folding into unique tertiary structures.Aptamer-based microfluidic platforms offer new ways to enhance the purity and capture efficiency of CTCs and EVs by combining the advantages of microfluidic chips as isolation platforms and aptamers as recognition tools.In this review,we first briefly introduce some new strategies for aptamer discovery based on traditional and aptamer-based micro-fluidic approaches.Then,we subsequently summarize the progress of aptamer-based microfluidics for CTC and EV detection.Finally,we offer an outlook on the future directional challenges of aptamer-based microfluidics for circulating targets in clinical applications.

Characterization of platelet-derived extracellular vesicles induced by differential platelet activation pathways / 中国输血杂志

【Objective】 To investigate the effects of platelet activation pathways on the characterization of platelet-derived extracellular vesicles(PEVs). 【Methods】 Whole blood from healthy donors was prepared into platelet suspension by centrifugation. Platelets were randomly divided into six groups, the ctrl group added no extra stimulus, and the other five groups were treated with collagen, adenosine diphosphate(ADP), thrombin, Ca2+ ionophore and freeze-thaw cycles (F-T) to activate platelets. Platelet-derived exosomes(PEXOs) and microvesicles(PMVs) were isolated by differential centrifugations, and then were determined by nano-flow cytometry and electron microscopy. The protein markers of PEXOs also were identified by western-blot. The protein concentration and content of PEXOs were also detected. 【Results】 CD9, CD81, TSG101 proteins were detected in all of the PEXOs, which had no calnexin. Both PEXOs and PMVs had CD41; PEXOs were cup-holder-like bilayer membrane vesicles under a transmission electron microscope, while PMVs were irregular membranous structure; 85%-95% of PEXOs were<100nm, 87%-94% of PMVs were 100-300nm. The concentration of exosomes and microvesicles in the F-T group was the highest(205.67±65.27 and 102.73±15.48), followed by the Ca2+ ionophore group(44.42±17.07 and 11.4±4.81). Although in the same size range, the numbers of PEVs induced by different activation conditions varied. The protein concentration of PEXOs in the F-T group(1.11±0.51) was higher than that in the control group(0.32±0.39), ADP group(0.41±0.31) and thrombin group(0.38±0.37), while the total protein(125.40±58.32) was higher than that in other three groups(the ctrl group 25.53±25.96 vs ADP group 37.21±15.73 vs thrombin group 36.28±24.18 vs Ca2+ ionophore group 47.09±23.29). 【Conclusion】 The biological characteristics of PEVs are affected by platelets activation pathways, whose ability to induce protein-packing into exosomes may also be relevant to the particle size. The freeze-thaw cycles can induce high concentrations of extracellular vesicles, which may be an ideal method for the preparation of PEVs.

Amino acid metabolism of fibroblasts involved in the occurrence and development of pulmonary fibrosis / 药学学报

Pulmonary fibrosis is a common pathological change in many chronic lung diseases, and its pathogenesis and characteristics are mainly caused by repeated lung alveolar injury leading to abnormal activation of fibroblasts and the accumulation of large amounts of extracellular matrix (ECM) deposition. Fibroblasts are not only responsible for constituting the interstitial structure of the lung but are also involved in the post-injury repairment in healthy lung tissue. In contrast, fibroblasts show a typical pro-fibrotic metabolic phenotype after differentiation into myofibroblasts during the development of pulmonary fibrosis. To synthesis large amount of collagen, the myofibroblasts have a strong metabolism characteristic of serine/glycine, glutamine, proline, and arginine. At the same time, the myofibroblast get the ability to resist cell apoptosis. As an important cell type for collagen degradation, fibroblasts reuse the amino acids of collagen to maintain cell metabolism. However, the myofibroblasts cannot degrade the ECM due to the suppression of autophagy activity, thus accelerating the progression of pulmonary fibrosis. This review attempts to summarize how amino acid metabolism of fibroblasts influence the pulmonary fibrosis.

Cross - species regulation and underlying mechanisms of parasite - derived non-coding RNAs: a review / 中国血吸虫病防治杂志

Parasite-derived non-coding RNAs (ncRNAs) not only contribute to life activities of parasites, and microRNA (miRNA), long non-coding RNA (lncRNA), and circular RNA (circRNA) may generate a competitive endogenous RNA (ceRNA) regulatory network with host miRNAs and mRNAs via extracellular vesicles, thereby participating in infection and pathogenic processes. This article presents an overview of characterizing ncRNAs derived from parasites and the cross-species regulatory role of parasite-derived ncRNAs in host gene expression and its underlying mechanisms.

Intervention effects and mechanism of Toddalia asiatica on cardiovascular damage in rats with collagen-induced arthritis / 中国药房

OBJECTIVE To investigate the intervention effects and potential mechanism of Miao medicine Toddalia asiatica on cardiovascular damage in rats with collagen-induced arthritis （CIA） based on vitamin D （VD） and neutrophil extracellular traps （NETs）. METHODS SD rats were randomly divided into the normal group （9 rats） and the modeling group （61 rats）. CIA model was prepared by multi-point injection of type Ⅱ bovine collagen+Fisher’s incomplete adjuvant； the model rats were randomly divided into the model group， methotrexate group （positive control， 1.5 mg/kg， twice a week）， vitamin D group [pathway validation， 1 000 IU/（kg·d）， once a day]， T. asiatica low-dose， medium-dose and high-dose groups [0.54， 1.08， 2.16 g/（kg·d）， calculated by crude drug， once a day]， with 9 rats in each group； they were given relevant medicine intragastrically for 4 consecutive weeks. Arthritis index scoring was performed after modeling and before administration， and plantar thickness was measured before and after the last administration； the histopathological changes of ankle joint， heart and abdominal aorta were observed in rats； the serum contents of myeloperoxidase （MPO）， interleukin-6 （IL-6） and 25-hydroxyvitamin D3 [25（OH）D3] were detected； the expressions of peptidylarginine deiminase 4 （PAD4）， NETs markers [citrullinated histone H3（CitH3）， MPO]， VD-related indicators [vitamin D receptor （VDR）， 25-hydroxyvitamin D-1α-hydroxylase （CYP27B1）] and IL-6 were determined in cardiac tissue. RESULTS Compared with the normal group， the plantar thickness of the arthritis index increased significantly in the model group （P＜0.01）. The obvious inflammatory cell infiltration and fibrous tissue hyperplasia were found in the ankle joint， the obvious myocardial fiber （No.2019YFC171250101） vacuoles and thickening of some surrounding blood vessel walls were found in the heart tissue， and the endothelial detachment was found in the abdominal aorta. The contents of MPO and IL-6 in serum increased significantly（P＜0.01），while the level of 25（OH）D3 decreased significantly （P＜0.01）； the protein expressions of PAD4， CitH3， MPO and IL-6 in myocardial tissue up-regulated significantly （P＜0.01）， while protein expression of VDR and CYP27B1 changed to acertain extent without significance （P＞0.05）. Compared with the model group， the pathological changes of ankle joints and cardiac tissue in rats were significantly improved in administration groups， and the above indicators were generally reversed （P＜0.05 or P＜0.01）. CONCLUSIONS T. asiatica can improve rheumatoid arthritis symptoms and cardiovascular damage by inhibiting the formation of NETs and inflammatory response， the mechanism of which may be associated with the regulation of VD expression.