Regenerative endodontic therapy based on tissue engineering mediating by inflammatory microenvironment / 中国组织工程研究

BACKGROUND:Recently,regenerative endodontic therapy is a promising alternative to the maturation of tissue engineering.Inflammatory microenvironment plays a key role in regulating pulp regeneration. OBJECTIVE:To focus on the change in the inflammatory pulp microenvironment,the balance between inflammation and regeneration,and the research advances in tissue-engineered regenerative endodontic therapy within the context of the inflammatory microenvironment to provide a reference for future investigations into regenerative endodontic therapy. METHODS:We conducted a literature search on PubMed and CNKI using search terms"pulp regeneration,inflammation,regenerative endodontic therapy,tissue engineering"in Chinese and English for articles published between 2013 and 2023.The review finally included 61 relevant articles. RESULTS AND CONCLUSION:(1)The changes in the microenvironment of pulpitis involve a complex interplay of cellular and molecular reactions,which,as inflammation progresses,ultimately the microenvironment hinders tissue repair more than facilitates it.(2)Inflammation can promote dental pulp regeneration through stem cell recruitment and activate the complement system,but it can also hinder the regenerative process through immunosuppression and fibrosis.(3)Tissue engineering's three components(stem cells,growth factors,scaffold materials)collaborate to balance inflammation and regeneration,for example,by using interleukin-6 to regulate dental pulp stem cells and foster a regenerative environment.(4)Current research has been largely silent on infection and inflammation issues.The mechanisms underlying changes in the microenvironment of pulpitis are still not fully understood.One promising avenue for improving the clinical applicability of regenerative dental pulp therapy is to achieve precise regulation of the inflammatory-regeneration balance and create a regenerative microenvironment by synergistically leveraging the three elements of tissue engineering.However,this field of investigation still exhibits significant gaps in understanding,necessitating further exploration into innovative strategies for facilitating dental pulp regeneration under inflammation.As such,additional animal experimentation and randomized clinical trials are required to establish a robust foundation for the clinical practice of tissue engineering-based regenerative dental pulp therapy.

The role of glucose metabolism reprogramming and its targeted therapeutic agents in inflammation-related diseases / 药学学报

Cells undergo glucose metabolism reprogramming under the influence of the inflammatory microenvironment, changing their primary mode of energy supply from oxidative phosphorylation to aerobic glycolysis. This process is involved in all stages of inflammation-related diseases development. Glucose metabolism reprogramming not only changes the metabolic pattern of individual cells, but also disrupts the metabolic homeostasis of the body microenvironment, which further promotes aerobic glycolysis and provides favourable conditions for the malignant progression of inflammation-related diseases. The metabolic enzymes, transporter proteins, and metabolites of aerobic glycolysis are all key signalling molecules, and drugs can inhibit aerobic glycolysis by targeting these specific key molecules to exert therapeutic effects. This paper reviews the impact of glucose metabolism reprogramming on the development of inflammation-related diseases such as inflammation-related tumours, rheumatoid arthritis and Alzheimer's disease, and the therapeutic effects of drugs targeting glucose metabolism reprogramming on these diseases.

Overview of the main biological mechanisms linked to changes in periodontal ligament stem cells and the inflammatory microenvironment / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Periodontitis is a complex chronic inflammatory disease. The invasion of pathogens induces the inflammatory microenvironment in periodontitis. Cell behavior changes in response to changes in the microenvironment, which in turn alters the local inflammatory microenvironment of the periodontium through factors secreted by cells. It has been confirmed that periodontal ligament stem cells (PDLSCs) are vital in the development of periodontal disease. Moreover, PDLSCs are the most effective cell type to be used for periodontium regeneration. This review focuses on changes in PDLSCs, their basic biological behavior, osteogenic differentiation, and drug effects caused by the inflammatory microenvironment, to provide a better understanding of the influence of these factors on periodontal tissue homeostasis. In addition, we discuss the underlying mechanism in detail behind the reciprocal responses of PDLSCs that affect the microenvironment.

Effects of Jianpiyiqi Decoction on Proliferation,Invasion and Apoptosis of Human Liver Cancer Cell Huh-7 in Inflammatory Microenvironment / 世界科学技术-中医药现代化

Objective To explore the effect of JianpiYiqi Decoction on the proliferation,invasion and apoptosis of Human hepatoma Huh-7 cells under the constructed inflammatory microenvironment stimulated by lipopolysaccharide(LPS)and adenosine triphosphate(ATP),and to detect the interleukin-1β(IL-1β)and interleukin-18(IL-18)Expression level in cells.Methods Construction drug-containing serum of JianpiYiqi.Adopt LPS and ATP were added to the cell culture medium to stimulate hepatoma cells and construct the inflammatory microenvironment.The cells were divided into blank group,model group,VX-765 group(10 μmol·L-1),low concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction low dosedrug-containing serum),Medium concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction Medium dose drug-containing serum),and high concentration of traditional Chinese medicine group(15%JianpiYiqi Decoction high dose drug-containing serum).Adopt CCK-8 method was used to detect the proliferation level of cells in each group after cell intervention;The apoptosis rate of cells in each group was detected by Annexin V-FITC/PI method;Adopt Transwell method was used to detect the level of cell invasion in each group;adopt PI single staining was used to detect the cell cycle level in each group;Adopt ELISA method and Western blot method were used to detection of IL-1β and 1l-18 expression level in Huh-7 cells.Results Compared with the blank group,in the model group stimulated by LPS and ATP,the proliferation level,invasion level,IL-1β and IL-18 expression level of Huh-7 cells were higher(P<0.05),and the apoptosis level was lower(P<0.05).Compared with other groups,the Medium concentration of traditional Chinese medicine group and the high concentration of traditional Chinese medicine group could effectively inhibit the proliferation and invasion level of Huh-7 cells,block the cell proliferation cycle,reduce the cell survival rate(P<0.05),significantly induce apoptosis of Huh-7 cells(P<0.05),and reduce the expression levels of IL-1β and IL-18 in Huh-7 cells(P<0.05).Conclusion The drug-containing serum JianpiYiqi.The medium and high concentrations of JianpiYiqi Decoction drug-containing serum can inhibit the proliferation and invasion,block the cell cycle and induce apoptosis of human liver cancer Huh-7 cells by improving the inflammatory microenvironment in liver cancer Huh-7 cells.Its mechanism may be related to the inhibition of liver cancer cells.Analyze the levels of inflammatory factors IL-1β and IL-18 to achieve the therapeutic purpose of primary liver cancer.

Effect of fudosteine on lung cancer cells in an inflammatory microenvironment based on metabolomics / 药学学报

GC-MS metabolomics was used to investigate the effects of fudosteine on lung cancer A549 cells in an inflammatory microenvironment. Eleven metabolites (malic acid, isoleucine, lactose, galactinol, creatinine, gluconic acid, oleic acid, phosphate, S-carboxymethyl-L-cysteine, uridine and tagatose) were identified in the metabolomics results and could be used as biomarkers of fudosteine treatment. Pathway enrichment analysis showed that the metabolic pathways of amino acids including isoleucine, valine, leucine, glycine, serine and threonine were significantly altered, as were the metabolic pathways of carbohydrates such as galactose and pentose phosphate. Fudosteine significantly reduced the level of inflammatory factors in A549 cells and corrected the inflammatory microenvironment by interfering with the effects of amino acid metabolites and amino acid metabolism pathways. This study reveals that fudosteine may be able to inhibit the continuous inflammatory response and prevent the further progression of lung cancer by suppressing the inflammatory microenvironment.

Application of reactive soft tissue preservation in extraction sockets of periodontitis: recent progress and perspectives / 口腔疾病防治

@#Chronic periodontitis is a prevalent disease; if left untreated, it is a main indication for tooth extraction and can lead to tooth loss. The reactive soft tissue, formed as a result of the immune response to chronic inflammation, is left in the compromised socket. The major concern is how to deal with the residual reactive soft tissue. Conservative thought states that the reactive soft tissue should be completely debrided. In addition, novel practices concerning the reactive soft tissue were proposed in recent trials, which demonstrated that there might be merits for soft and hard tissue regeneration with preservation of the reactive soft tissue. Studies have shown that mesenchymal stem cells exist in inflammatory reactive soft tissue, stressing their potential in tissue regeneration. Although the therapeutic value is highly promising, the specific components of the reactive soft tissue and the standard on whether it should be preserved need further investigation.

Cognition of Ovarian Cancer Microenvironment in Traditional Chinese Medicine from Perspective of "Yin-Yang Balance" / 中国实验方剂学杂志

Ovarian cancer， one of gynecological malignancies， is often diagnosed at the late stage because of the atypical early symptoms and has become a major killer of women. Research has found that the co-evolution of tumor cells and their surrounding microenvironment is an important cause for the occurrence and development of ovarian cancer. It is believed in traditional Chinese medicine （TCM） that Yin and Yang are the roots of everything， and their balance， namely Yin being at peace and Yang being compact （“Yin Ping Yang Mi”） is a sign of good health. The mutual opposition， restriction， and rooting of Yin and Yang as well as their waning and waxing and transformation are the keys to maintaining the balance. In TCM， ovarian cancer falls into the category of abdominal mass， which results from the struggle between healthy Qi and evil Qi. When the healthy Qi is deficient and the evil Qi is excessive， the balance between Yin and Yang will be destroyed， triggering the body and ovarian cancer microenvironment as well as the relevant factors in the inflammatory microenvironment to be mutually opposed， restricted， and transformed， highly consistent with the dynamic development of Yin and Yang. At present， the studies concerning TCM intervention in the inflammatory microenvironment of ovarian cancer mostly focus on the signaling pathways to reveal the advantages of TCM multiple components against cancer cells via multiple targets， but they fail to explain the TCM efficacy from the perspective of Yin-Yang balance. Therefore， guided by the concept of Yin-Yang balance， this paper macroscopically and microscopically explored the effects of the changed factors in inflammatory microenvironment on the occurrence and development of ovarian cancer， and put forward that the prevention and control principles of ovarian cancer should lie in the "adjustment of Yin-Yang balance"， accompanied by healthy Qi reinforcement and pathogen elimination. This paper has laid the foundation for the elucidation of modern research achievements regarding the ovarian cancer microenvironment with TCM theory and provide a theoretical basis for the clinical treatment of ovarian cancer with integrated TCM and western medicine.

Effects of manual loading on calcitonin gene-related peptide and nerve growth factor in rats with chronic low back pain / 中国骨伤

OBJECTIVE@#To observe the analgesic effect of manipulation loading on chronic low back pain (CLBP) model rats and the expression of inflammatory factors in psoas major muscle tissue, and to explore the improvement of manipulation on local inflammatory microenvironment.@*METHODS@#Thirty two SPF male SD rats weighing 340-360g were randomly divided into blank group, sham operation group, chronic low back pain model group and treatment group, with 8 rats in each group. In the model group, L@*RESULTS@#There was no significant difference in PWT and PWL between the blank group and the sham operation group after modeling (@*CONCLUSION@#Local massage loading has analgesic effect on CLBP rats, at the same time, it can inhibit the content of CGRP and NGF in psoas muscle tissue of CLBP rats, and improve the local inflammatory microenvironment.

Effect of macrophage dysfunction on diabetic wound healing: a review of research progress / 中华显微外科杂志

Macrophages play a crucial role in inflammatory, proliferative and reconstructive phases in wound healing. A dysfunction of macrophage could lead to a delay of healing. As the most common type of chronic wounds, the diabetic wound may ultimately result in a delayed or failed wound healing due to a high glucose microenvironment and abnormal metabolic environment. Such abnormal metabolic environment may lead to the aberrant macrophage polarization, abnormal secretion of cytokines and aberrant phagocytic function hence cause prolonged inflammation with excessive oxidative stress reaction. As the consequence, the inflammatory phase in diabetic wound is lengthy while the proliferative and reconstructive phases are usually delayed. The diabetic wound may result in enormous financial burden to patients, the healthcare and the society. This study briefly reviewed the recent research progresses at home and abroad, and analysed and summarized the roles of the dysfunction of macrophage polarization, secretion and phagocytosis in the process of diabetic wound healing.

Investigation of Mechanism of Tryptanthrin Inhibiting Breast Cancer in Mice Based on Proteomics / 中国实验方剂学杂志

Objective:A systematical study on the anti-breast cancer mechanism of tryptanthrin in breast cancer-bearing mice was done by Label-free proteomics. Method:UPLC-MS was used to detect the expressed-proteins of tryptanthrin inhibiting breast cancer in mice， chromatographic separation was achieved on the Ionoptics nano UPLC C18 column （0.075 mm×250 mm， 1.6 μm）， and gradient elution was performed with 0.1% formic acid aqueous solution-0.1% formic acid acetonitrile solution as mobile phase. Data acquisition was carried out in electrospray ionization （ESI） under the positive ion mode， the scanning range was m/z 100-1 700， MaxQuant 1.6.5.0 was used for database retrieval. Label-free proteomics with high resolution mass spectrometry was used to screen differentially expressed proteins between the model group of 4T1 breast cancer mice and oral administration group of tryptanthrin （100 mg·kg-1）. The proteomics of tryptanthrin against breast cancer was carried out. Result:A total of 3 997 proteins were identified in this proteomics research， and 2 911 proteins were quantifiable. A total of 750 differentially expressed proteins were identified between the model group and the tryptanthrin group， 286 proteins were up-regulated and 464 proteins were down-regulated. Gene ontology analysis showed that these differentially expressed proteins were mainly involved in biological processes of proliferation， cell migration， apoptosis， immunity， angiogenesis， inflammatory regulation， etc. Kyoto encyclopedia of genes and genomes pathway analysis further indicated that these proteins were mainly concentrated in T cell receptors， B cell receptors， Toll-like receptors， nuclear transcription factor-κB （NF-κB）， Ras proteins， interleukin-17， tumor necrosis factor， phosphatidylinositol 3-kinase/protein kinase B （PI3K-Akt）， mitogen-activated protein kinase （MAPK） and other signaling pathways. Conclusion:The differentially expressed proteins closely related to anti-breast cancer effect of tryptanthrin on 4T1 breast cancer mice are effectively screened out， including up-regulating proteins of leukocyte differentiation antigen 14 （CD14）， prostaglandin G/H synthase 2 （PTGS2）， E3 ubiquitin-protein ligase and down-regulating proteins of CD44， heat shock 70 kDa protein 1A （HSPA1A）， macrophage migration inhibitory factor （MIF）， NF-κB， ribosomal protein S6 kinase alpha-4 （RPS6KA4） and high mobility group protein B1 （HMGB1）. These findings suggest that tryptanthrin can inhibit breast cancer in mice mainly through regulating tumor inflammatory microenvironment.

Progress in Metabolic Reprogramming of Tumor-associated Macrophages in Tumor Microenvironment / 中国实验方剂学杂志

Occupying more than half of the tumor volume in a variety of solid tumors, tumor-associated macrophages (TAMs) are an important part of the tumor microenvironment (TME) with high plasticity and heterogeneity. In the early stages of tumor development, TAMs mediate antitumor effect through phagocytosis and their antioxidant functions. However, in order to meet the needs of self-renewal and proliferation, malignant tumor cells continuously adjust their metabolic patterns, leading to the accumulation of metabolites such as lactate, reactive oxygen species, nitric oxide, arachidonic acid and prostaglandin in the TME, which results in the changes in its inflammatory profiles, thereby altering the metabolism and function of TAMs and ultimately promoting the tumor development. Therefore, further understanding of the metabolism and immune responses of TAMs in the TME during tumor progression is warranted and the investigation may lead to identification of novel potential targets for cancer immunotherapy. This review aims to clarify the close relationship between TAMs metabolism and TME immune response, to reveal the mechanism of tumor immunosuppression produced by TAMs metabolism, and to provide new treatment ideas and approaches for tumor immunotherapy.

Effect of Xihuangwan on NLRP3 Inflammatory Bodies and Their Products and Tumor Proliferation of Lung Cancer A549 Bearing Nude Mice in Inflammatory Microenvironment / 中国实验方剂学杂志

Objective:To study the antitumor effect of Xihuangwan on A549 lung cancer nude mice in inflammatory microenvironment, and explore the effect of Xihuangwan on the expressions of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammatory bodies and their products in serum and tumor tissue of A549 lung cancer nude mice. Method:The lung cancer A549 cell model was established in nude mice with lung cancer, and the lung cancer A549 cell model was established in inflammatory microenvironment by adding lipopolysaccharide (LPS) + adenosine triphosphate (ATP) to the culture medium. After modeling, the rats were randomly divided into blank group (equal volume of normal saline), positive drug control group (MCC950 solution, 0.79 g·kg-1), and low, medium, high-dose Xihuangwan groups (0.39, 0.78, 1.95 g·kg-1). The rats were administered orally once a day for 21 days, and then sacrificed. The tumor tissues were stripped to measure the tumor body. The expressions of NLRP3, malondialdehyde（MDA）, interleukin (IL)-1β, IL-18 and NLRP3 were detected by Western blot, and the levels of IL-1β, IL-18 and MDA were detected by enzyme linked immunosorbent assay（ELISA）. Result:Compared with the blank group, the tumor inhibition rates in the positive drug control group and the low, medium and high dose Xihuangwan group were 39.21%, 31.72%, 42.24% and 55.68%. ELISA showed that the high-dose Xihuangwan group could significantly reduce the expressions of MDA, IL-1β and IL-18 in the serum of nude mice (P<0.05). Western blot showed that the high-dose Xihuangwan group could effectively reduce the protein expressions of MDA, IL-1β, IL-18 and NLRP3 in tumor of nude mice (P<0.05), the results of immunohistochemistry showed that the expression rate of NLRP3 in the tumor tissues of nude mice was reduced in the positive drug group and each dose of Xihuangwan group (P<0.05). Conclusion:Xihuangwan can inhibit the growth of tumor tissue of A549 cells bearing lung cancer in nude mice. The mechanism may be that it can inhibit the growth of tumor cells by inhibiting the expression of NLRP3 inflammatory bodies, IL-1β, IL-18, MDA tables, and then inhibiting the inflammatory microenvironment of tumor cells.

Effect of Testudinis Carapax et Plastrum Aqueous Extract Regulating NF-κB Inflammation Microenvironment on Osteogenic Differentiation of MC3T3-E1 / 中国实验方剂学杂志

Objective::To investigate the role and mechanism of Testudinis Carapax et Plastrum aqueous extract in promoting osteogenic differentiation of mouse preosteoblast cell line(MC3T3-E1) by regulating nuclear transcription factor-κB(NF-κB) inflammation microenvironment. Method::MC3T3-E1 cells were cultured in vitro, and osteogenic induction (OI) was performed. Testudinis Carapax et Plastrum was prepared and treated the cells. Cells were devided into control group, osteogenic induction group and Testudinis Carapax et Plastrum (20 mg·L-1)with osteogenic induction group. The proliferation of MC3T3-E1 was detected by cell counting kit-8 (CCK-8), and the optimum concentration of intervention was determined. MC3T3-E1 differentiation and osteogenic mineralization were assayed using alkaline phosphatase (ALP) and Alizarin red staining (ARS), respectively. The expressions of NF-κB p65, NF-κB p105, interleukin-6(IL-6), ALP and Collagen-Ⅰ(COL-Ⅰ) mRNA were detected by Real-time PCR. Result::The results of CCK-8 showed that the proliferation of MC3T3-E1 did not change statistically with time, but it showed an upward trend, while the proliferation at 20 mg·L-1 was more obvious than other groups. The ALP and ARS showed that the positive staining rate of osteogenic induction group and Testudinis Carapax et Plastrum with osteogenic induction group were higher than control group.Real-time PCR results showed that on the 7th day in culture, the expression of NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01), and the expression of ALP and COL-Ⅰ mRNA was significantly upregulated(P<0.05), on the 14th day, the expression of NF-κB p65, NF-κB p105 and IL-6 mRNA in Testudinis Carapax et Plastrum with osteogenic induction group was significantly lower than that in control group (P<0.01). The expression of ALP and COL-Ⅰ mRNA was significantly increased (P<0.05, P<0.01). Conclusion::Testudinis Carapax et Plastrum aqueous extract can promote osteogenic differentiation of MC3T3-E1 via a mechanism associated with the regulation of inhibition of NF-κB inflammatory microenvironment.

Macrophages′ influence on preadipocytes in the inflammatory microenvironment of adipose tissue / 中华整形外科杂志

Fat regeneration plays an important role in maintaining fat volume after transplantation, and preadipocyte is the primary source of fat regeneration. Inflammatory microenvironment plays an important role in regulating fat regeneration. As a key role in this process, the effect of macrophages on preadipocytes is a research hotspot. In this paper, we summarized the previous studies in related fields at home and abroad, and made a briefly review of the effects of macrophages on adipogenesis, proliferation and survival of preadipocytes.

Endoplasmic reticulum-mitochondrial contact regulates osteogenic differentiation of periodontal ligament stem cells via mitofusion 2 in inflammatory microenvironment / 中华口腔医学杂志

Objective@#To investigate the effect of endoplasmic reticulum (ER)-mitochondria coupling and the expression of mitofusion 2 (Mfn2) in periodontal ligament stem cells (PDLSC), so as to provide a theoretical basis and therapeutic target for promoting periodontal regeneration in treatment of periodontitis.@*Methods@#The periodontal ligament tissue was scraped from extracted intact human teeth and teeth with periodontitis collected in the Department of Oral and Maxillofacial Surgery, the Fourth Military Medical University. The health PDLSC (H-PDLSC) and inflammatory PDLSC (P-PDLSC) were obtained respectively from the primary culture of the human teeth and cloned using 1imited diluted method. The level of ER-mitochondrial coupling was observed by transmission electron microscopy and organelle-specific fluorescence staining. Quantitative real-time PCR (qPCR) was used to detect the expression of Mfn2 in H-PDLSC and P-PDLSC. Tumor necrosis factor alpha (TNF-α) was used to simulate the inflammatory microenvironment. H-PDLSC was cultured in normal medium and media containing 5 and 10 mg/L TNF-α, named as H-PDLSC group, H-PDLSC+TNF-α (5 mg/L) group and H-PDLSC+TNF-α (10 mg/L) group, respectively. At the 7th day, qPCR was applied to detect the mRNA level of Mfn2. The expression of Mfn2 in P-PDLSC was down-regulated by small interfering RNA siMfn2. The osteogenic differentiation of P-PDLSC and P-PDLSC+siMfn2 were examined by qPCR at the 7th day, and alizarin red staining and cetyl pyridine chloride quantitative analysis at the 28th day after osteogenic induction.@*Results@#The level of ER-mitochondrial coupling in the P-PDLSC group (the length of the coupling structure/mitochondrial perimeter was 0.55±0.10, the length of the coupling structure/endoplasmic reticulum perimeter was 0.44±0.08) was significantly higher than that in the H-PDLSC group (P<0.01). The co-localization of endoplasmic reticulum and mitochondria of P-PDLSC group was 0.71±0.09, which was significantly higher than that of H-PDLSC group (P<0.01). The expression level of Mfn2 in P-PDLSC (1.46±0.10) was higher than that in H-PDLSC (0.99±0.08). The expression levels of Mfn2 in H-PDLSC+TNF-α (5 mg/L) and H-PDLSC+TNF-α (10 mg/L) groups were 1.28±0.19, 1.54±0.43, respectively, which were both significantly higher than that in H-PDLSC (0.82±0.14) (P<0.01). P-PDLSC transfected with siMfn2 down-regulated the expression of Mfn2, and the osteogenic differentiation ability of P-PDLSC was restored. The results showed that the expression of alkaline phosphatase, Runt-related transcription factor-2 (RUNX2) and osteocalcin mRNA in P-PDLSC+siMfn2 group were significantly higher than that of the control group (P<0.01). The alizarin red staining and quantitative results of cetyl pyridinium chloride were consistent with the qPCR results.@*Conclusions@#In the microenvironment of inflammation, ER-mitochondrial coupling and the expression of Mfn2 of PDLSC increased, which might lead to a decrease in osteogenic differentiation of PDLSC. The specific mechanism needs to be further studied.

Effects of Xiaoyan Decoction on TGF-β/Smad3/MMP-9 signaling pathway in SP cells / 中草药

Objective: To investigate the specific mechanism of Xiaoyan Decoction in the metastasis of non-small cell lung cancer. Methods: The detection method of Western blotting method and quantitative PCR Xiaoyan Decoction intervention expression of many inflammatory proteins and cytokines in the lower lung cancer stem Xiaoyan Decoction explore the possible mechanism of effect of non small cell lung cancer metastasis. Results: Western blotting detection showed that the expression of IL-6, TGF-β, p-Smad3, and MMP-9 protein in both experimental group and control group decreased significantly (P < 0.05), the difference was statistically significant; RT-PCR showed that the experimental group compared with the control group IL-6, TGF-β, TNF-α, MMP-2, alpha beta MMP-9 gene expression decreased significantly (P < 0.05), the difference was statistically significant. Conclusion: Xiaoyan Decoction mainly affects the metastasis of non-small cell lung cancer by inhibiting the transmission of TGF-β/Smad3 signaling pathway in the inflammatory microenvironment of lung cancer.

IL-6 and TNF-α promote the differentiation of human BMSCs to TFAs by mimicry inflammatory microenvironment / 基础医学与临床

Objective To observe whether BMSCs differentiate into TAFs in inflammatory microenvironment simula-ted by IL-6 and TNF-α.Methods The experiment was divided into 9 groups:the BMSCs group,the 50 ng/mL, 100 ng/mL IL-6 intervention group, the 50 ng/mL,100 ng/mL TNF-α intervention group, the 50 ng/mL IL-6+50 ng/mL TNF-α intervention group,the 50 ng/mL IL-6+100 ng/mL TNF-α intervention group,the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group and the 100 ng/mL IL-6+100 ng/mL TNF-α intervention group; After continuous induction for 42 days, BMSCs cell morphology and cycle, TAFs-tagged α-SMA and FAP protein were examined by phase-contrast microscope,flow cytometry and Western blot method. Results Compared with the normal control group, the 100 ng/mL IL-6+50 ng/mL TNF-α intervention group BMSC cell proliferation was significantly promoted;G1phase decreased in proportion and S phase increased;α-SMA and FAP protein expression was signifi-cantly increased (P＜0.05).Conclusions Microenvironment simulated by the 100 ng/mL IL-6+50 ng/mL TNF-α may induce the abnormal change of the BMSCs morphological and proliferative characteristics. TAFs reference mole-cule α-SMA and FAP expression is increased.

Effects of inflammatory microenvironment mediated by macrophage on the proliferation and osteogenic differentiation of periodontal ligament cells / 口腔疾病防治

Objective@#The present study investigated the effects of the inflammatory microenvironment mediated by macrophages on the proliferation and osteogenic differentiation of periodontal ligament cells (PDLCs).@*Methods@#Conditioned medium containing inflammatory factors was collected following macrophage activation with 1 μg/mL lipopolysaccharide (LPS). PDLCs were isolated from healthy teeth and cultured in conditioned medium (LPS-CM group) or normal medium (control group), and the proliferation of PDLCs was detected using the MTT assay. The cells were cocultured with an osteogenic inducer for 3 d and 7 d, and the alkaline phosphatase (ALP) activity of PDLCs was detected using an ALP kit. The mRNA expression levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and collagen I (COL-I) were detected using real-time PCR, and the protein levels of RUNX2, OCN, and COL-I were detected using Western blotting. Mineralization nodules were observed using Alizarin red staining after osteoinduction for 14 d. @*Results@#The OD value of PDLCs in the LPS-CM group was lower than that in the control group (P < 0.05). The mRNA levels of RUNX2, OCN, and COL-I in the LPS-CM group were lower than those in the control group (P < 0.05). In addition to the OCN 3 d group (t = 2.75, P = 0.056), the protein expression of RUNX2, OCN, and COL-I in the LPS-CM group was lower than that in the control group (P < 0.05). However, the ALP activity of the LPS-CM group was higher than that of the control group, which was 1.58-fold greater (t = 5.91, P = 0.030) at 3 d and 1.29-fold greater (t = 6.01, P = 0.046) at 7 d. The number of calcified nodules in the LPS-CM group was significantly less than that in the control group (t = 8.63, P = 0.048). @*Conclusion@# The inflammatory microenvironment mediated by macrophages may inhibit the proliferation and osteogenic differentiation of PDLCs.

Mitofusin-1 regulates osteogenic differentiation of periodontal ligament stem cells / 实用口腔医学杂志

Objective: To detect the expression of mitofusion-1(Mfn1) in periodontal ligament stem cells (PDLSCs) isolated from healthy and periodontitis tissue and to study the effect of Mfn1 on the osteogenic differentiation of PDLSCs. Methods: PDLSCs were isolated from the healthy and periodontitis human samples(H-PDLSCs and P-PDLSCs). IL-1β was applied to mimic the inflammation microenvironment(H-PDLSCs + IL-1β). RT-PCR was used to detect the expression of Mfn1 in HPDLSCs, P-PDLSCs and H-PDLSCs + IL-1β. The expression of Mfn1 in P-PDLSCs was down-regulated by siRNA of Mfn1 (siMfn1). The osteogenic differentiation of the cells was examined by RT-PCR, alizarin red staining and cetyl pyridine chloride quantitative analysis. Results: The expression level of Mfn1 in P-PDLSCs and H-PDLSCs + IL-1β (5 μg/ml) groups was higher than that in H-PDLSCs(P＜ 0. 05). When the expression of Mfn1 in P-PDLSCs was down-regulated by siMfn1 the osteogenic differentiation ability of P-PDSLCs was restored(P＜ 0. 05). Conclusion: Inflammation may promote Mfn1 expression in PDLSCs and inhibite the osteogenic differentiation of P-PDLSCs.

Effect of PIAS1 on Epithelial-mesenchymal Transition of Gastric Cancer under Inflammatory Microenvironment / 胃肠病学

Background:Protein inhibitor of activated STAT 1( PIAS1 )is an important regulator for inflammatory signaling network,which is low expressed in gastric cancer and associated with development of cancer,but its mechanism has not been elucidated. Aims:To investigate the effect of PIAS1 on epithelial-mesenchymal transition( EMT)of gastric cancer under inflammatory microenvironment. Methods:Recombinant adenovirus Ad5/F35-PIAS1 and Ad5/F35-null were constructed and transfected into gastric cancer cell line SGC-7901,mRNA and protein expressions of PIAS1 were detected by RT-PCR and Western blotting,respectively. SGC-7901 cells were divided into IL-6 treatment group,Ad5/F35-PIAS1﹢IL-6 treatment group and Ad5/F35-null﹢IL-6 treatment group. Cell proliferation was measured by MTT method,migration and invasion capacities were assessed by wound healing test and Transwell chamber invasion assay. Protein expressions of E-cadherin,Snail,Twist,Vimentin and P-p38MAPK were assessed by Western blotting. Results:The transfection of Ad5/F35-PIAS1 significantly increased the expressions of PIAS1 mRNA and protein in SGC-7901 cells. Compared with IL-6 treatment group and Ad5/F35-null﹢IL-6 treatment group,capacities of cell proliferation,migration and invasion were significantly decreased(P ﹤0. 01);protein expressions of Snail,Twist,Vimentin and P-p38MAPK were significantly decreased while expression of E-cadherin protein was significantly increased in Ad5/F35-PIAS1 ﹢IL-6 treatment group ( P﹤0. 01). No significant differences in above-mentioned indices were found between IL-6 treatment group and Ad5/F35-null﹢IL-6 treatment group(P﹥0. 05). Conclusions:PIAS1 could inhibit EMT of gastric cancer cells under inflammatory microenvironment,and may play an important role in inhibition of tumor invasion and metastasis.