Up-regulation of GINS1 highlighted a good diagnostic and prognostic potential of survival in three different subtypes of human cancer / A regulação positiva de GINS1 destacou um bom potencial diagnóstico e prognóstico de sobrevivência em três subtipos diferentes de câncer humano

Abstract Cancer is a fatal malignancy and its increasing worldwide prevalence demands the discovery of more sensitive and reliable molecular biomarkers. To investigate the GINS1 expression level and its prognostic value in distinct human cancers using a series of multi-layered in silico approach may help to establish it as a potential shared diagnostic and prognostic biomarker of different cancer subtypes. The GINS1 mRNA, protein expression, and promoter methylation were analyzed using UALCAN and Human Protein Atlas (HPA), while mRNA expression was further validated via GENT2. The potential prognostic values of GINS1 were evaluated through KM plotter. Then, cBioPortal was utilized to examine the GINS1-related genetic mutations and copy number variations (CNVs), while pathway enrichment analysis was performed using DAVID. Moreover, a correlational analysis between GINS1 expression and CD8+ T immune cells and a the construction of gene-drug interaction network was performed using TIMER, CDT, and Cytoscape. The GINS1 was found down-regulated in a single subtypes of human cancer while commonly up-regulated in 23 different other subtypes. The up-regulation of GINS1 was significantly correlated with the poor overall survival (OS) of Liver Hepatocellular Carcinoma (LIHC), Lung Adenocarcinoma (LUAD), and Kidney renal clear cell carcinoma (KIRC). The GINS1 was also found up-regulated in LIHC, LUAD, and KIRC patients of different clinicopathological features. Pathways enrichment analysis revealed the involvement of GINS1 in two diverse pathways, while few interesting correlations were also documented between GINS1 expression and its promoter methylation level, CD8+ T immune cells level, and CNVs. Moreover, we also predicted few drugs that could be used in the treatment of LIHC, LUAD, and KIRC by regulating the GINS1 expression. The expression profiling of GINS1 in the current study has suggested it a novel shared diagnostic and prognostic biomarker of LIHC, LUAD, and KIRC.

Resumo O câncer é uma doença maligna fatal e sua crescente prevalência mundial exige a descoberta de biomarcadores moleculares mais sensíveis e confiáveis. Investigar o nível de expressão de GINS1 e seu valor prognóstico em cânceres humanos distintos, usando uma série de abordagens in silico em várias camadas, pode ajudar a estabelecê-lo como um potencial biomarcador de diagnóstico e prognóstico compartilhado de diferentes subtipos de câncer. O mRNA de GINS1, a expressão da proteína e a metilação do promotor foram analisados ​​usando UALCAN e Human Protein Atlas (HPA), enquanto a expressão de mRNA foi posteriormente validada via GENT2. Os valores prognósticos potenciais de GINS1 foram avaliados por meio do plotter KM. Em seguida, o cBioPortal foi utilizado para examinar as mutações genéticas relacionadas ao GINS1 e as variações do número de cópias (CNVs), enquanto a análise de enriquecimento da via foi realizada usando DAVID. Além disso, uma análise correlacional entre a expressão de GINS1 e células imunes T CD8 + e a construção de uma rede de interação gene-droga foi realizada usando TIMER, CDT e Cytoscape. O GINS1 foi encontrado regulado negativamente em um único subtipo de câncer humano, enquanto comumente regulado positivamente em 23 outros subtipos diferentes. A regulação positiva de GINS1 foi significativamente correlacionada com a sobrevida global pobre (OS) de Carcinoma Hepatocelular de Fígado (LIHC), Adenocarcinoma de Pulmão (LUAD) e Carcinoma de Células Claras Renais de Rim (KIRC). O GINS1 também foi encontrado regulado positivamente em pacientes LIHC, LUAD e KIRC de diferentes características clínico-patológicas. A análise de enriquecimento de vias revelou o envolvimento de GINS1 em duas vias diversas, enquanto poucas correlações interessantes também foram documentadas entre a expressão de GINS1 e seu nível de metilação do promotor, nível de células imunes T CD8 + e CNVs. Além disso, também previmos poucos medicamentos que poderiam ser usados ​​no tratamento de LIHC, LUAD e KIRC, regulando a expressão de GINS1. O perfil de expressão de GINS1 no estudo atual sugeriu que é um novo biomarcador de diagnóstico e prognóstico compartilhado de LIHC, LUAD e KIRC.

Effects of tumor derived exosomes on T cells markers expression / Efeitos de exossomos derivados de tumor na expressão de marcadores de células T

Exosomes are 30-120nm bio particles transferred from donor to recipient cells leading to modification in their regulatory mechanisms depending upon the coded message in the form of loaded biomolecule. Cancer cells derived exosomes the true representatives of the parent cells have been found to modify the tumor surrounding/distinct regions and participate in metastasis, angiogenesis and immune suppression. Tis study was aimed to study the effects of tumor mice derived exosomes on the normal mice spleen isolated T cells by using co-culture experiments and flow cytometer analysis. We mainly focused on some of the T cells population and cytokines including IFN-γ, FOXP3+ regulatory T (Treg) cells and KI67 (proliferation marker). Overall results indicated random changes in different set of experiments, where the cancer derived exosomes reduced the IFN-γ expression in both CD4 and CD8 T cells, similarly the Treg cells were also found decreased in the presence of cancer exosomes. No significant changes were observed on the Ki67 marker expression. Such studies are helpful in understanding the role of cancer exosomes in immune cells suppression in tumor microenvironment. Cancer exosomes will need to be validated in vivo and in vitro on a molecular scale in detail for clinical applications.

Os exossomos são biopartículas de 30-120 nm transferidas de células doadoras para células receptoras, levando à modificação em seus mecanismos reguladores, dependendo da mensagem codificada na forma de biomolécula carregada. Verificou-se que exossomos derivados de células cancerosas ­ os verdadeiros representantes das células-mãe ­ modificam as regiões circundantes / distintas do tumor e participam da metástase, angiogênese e imunossupressão. Este estudo teve como objetivo estudar os efeitos de exossomos derivados de camundongos com tumor nas células T isoladas de baço de camundongos normais, usando experimentos de cocultura e análise de citômetro de fluxo. Concentrou-se, principalmente, em algumas populações de células T e citocinas, incluindo IFN-γ, células T reguladoras FOXP3 + (Treg) e KI67 (marcador de proliferação). Os resultados gerais indicaram mudanças aleatórias em diferentes conjuntos de experimentos, em que os exossomos derivados de câncer reduziram a expressão de IFN-γ em células T CD4 e CD8, da mesma forma que as células Treg também foram encontradas diminuídas na presença de exossomos de câncer. Nenhuma mudança significativa foi observada na expressão do marcador Ki67. Esses dados são úteis para a compreensão do papel dos exossomos do câncer na supressão de células do sistema imunológico no microambiente tumoral. Exossomos de câncer precisarão ser validados in vivo e in vitro em escala molecular com detalhes para aplicações clínicas.

Phytochemical profile and biological activities of Bromelia antiacantha extracts / Perfil fitoquímico e atividades biológicas de extratos de Bromelia antiacantha

Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

Effect of Ermiaosan on Expression of Th17/Treg Cells in DBA/1 Mice with Collagen-induced Arthritis / 中国实验方剂学杂志

ObjectiveTo observe the effect of classic prescription Ermiaosan （EMS） on the differentiation of T helper 17 （Th17） /regulatory T （Treg） cells in collagen-induced arthritis （CIA） DBA/1 mice. MethodDBA/1 mice were randomized into normal group， CIA group， EMS （5.4 g·kg-1） group， and methotrexate （MTX，0.5 mg·kg-1） group according to the body weight. DBA/1 mice in CIA group， EMS group， and MTX group were immunized with equal volume of bovine type Ⅱ collagen and complete Freund's adjuvant on the first day and were immunized with equal volume of bovine type Ⅱ collagen and incomplete Freund's adjuvant on the 21st day to induce CIA. On the day of the secondary immunization， intragastric administration started and lasted 28 days （three times/week for MTX group， and once/day for other groups）. The symptoms of CIA mice such as joint redness and swelling were observed from the 22nd day， and the arthritis was scored. After the sampling on the 49th day， synovitis of CIA mice was observed based on hematoxylin-eosin （HE） staining. Double-labeling immunofluorescence （IF） method was used to detect the expression of Th17 cell marker IL-17 and Treg cell marker forkhead transcription factor P3 （FoxP3） in CD4+T cells in CIA mouse joints. The proportion of Th17 and Treg cells in the spleen and lymph nodes of mice was detected by flow cytometry. ResultCompared with the normal group， CIA group had obvious synovitis， disordered joint structure， severely damaged articular cartilage and bone， serious bone erosion （P<0.01）， high Th17/Treg value in joint tissue （P<0.01）， high proportion of Th17 cells in spleen and lymph nodes （P<0.01）， and low proportion of Treg cells （P<0.01）. Compared with CIA group， EMS group and MTX group had normal joint structure， mild bone erosion and bone destruction， complete and smooth joint surface， low Th17/Treg value （P<0.01）， low proportion of Th17 cells in spleen and lymph nodes （P<0.01）， and high proportion of Treg cells in spleen and lymph nodes （P<0.01）. ConclusionEMS regulates the balance of Th17/Treg， inhibits the expression of Th17 cells， and promotes the expression of Treg cells in CIA mice， thereby exerting therapeutic effect on RA.

Prevention and Treatment of Diabetic Nephropathy by Regulating Endoplasmic Reticulum Stress with Traditional Chinese Medicine： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN） is one of the serious and common microvascular complications of diabetes mellitus （DM） and the main cause of end-stage renal disease （ESRD）. Endoplasmic reticulum stress （ERS） is a common stress defense mechanism in eukaryotic cells. In the ERS state， cells activate the unfolded protein response （UPR） to enhance the folding of unfolded proteins and the degradation of misfolded proteins， so as to restore the normal physiological function of the endoplasmic reticulum and avoid cell damage. However， excessive or chronic persistent ERS can induce apoptosis， inflammation， oxidative stress and other pathways to eventually cause cell damage. In recent years， a large number of studies have confirmed that ERS is closely associated with the occurrence and development of DN. In the case of DN， ERS is involved in the damage or protection of podocytes， glomerular mesangial cells， renal tubular epithelial cells， and glomerular endothelial cells. The regulation of ERS has become one of the hotspots in the prevention and treatment of DN and has received extensive attention in the field of traditional Chinese medicine. This paper systematically expounds the role of ERS in the occurrence and development of DN and summarizes the ERS-targeted regulation of DN by traditional Chinese medicine， with a view to providing certain research ideas for the prevention and control of DN with traditional Chinese medicine.

Research progress on the regulation of epithelial cell senescence by Sirt1 / 口腔疾病防治

@#In the process of enamel development, premature senescence and apoptosis of ameloblasts are important causes of hereditary enamel hypoplasia. Silence information regulator 2-related enzyme 1 (Sirt1) is a nicotinamide adenosine dinucleotide (NAD+)-dependent deacetylase that has been widely reported to be involved in the regulation of cell senescence. This paper reviews the research progress of Sirt1 regulating epithelial cell senescence, starting with the structural characteristics of Sirt1, and further expounds on the relationship between Sirt1 and senescence. When epithelial cells are stimulated, Sirt1 affects the senescence of epithelial cells in many ways, such as mitochondrial dysfunction. Sirt1 participates in regulating mitochondrial function and metabolic homeostasis, and telomere length is negatively related to senescence. Sirt1 regulates the expression of telomere reverse transcriptase needed for telomere extension, thus positively regulating telomere homeostasis. DNA damage will undergo damage repair, unrepaired DNA damage will cause cell senescence, and the Sirt1/p53 pathway can inhibit epithelial cell senescence by reducing DNA damage. Senescent cells are the source of chronic inflammation, and chronic inflammation can also promote aging in many ways. Sirt1 inhibits epithelial cell senescence by relieving inflammatory symptoms. In future research, we can focus on the effect of Sirt1 on ameloblast senescence and explore its specific mechanism of action on ameloblasts to find a breakthrough in the etiology and treatment of enamel hypoplasia.

Effect of mechano-growth factor on proliferation and differentiation of periodontal ligament stem cells / 口腔疾病防治

Objective@#To investigate the effects and molecular mechanisms of mechano-growth factor (MGF) on the proliferation and differentiation of periodontal ligament stem cells (PDLSCs).@* Methods@#PDLSCs were obtained using magnetic bead sorting. Flow cytometry was performed to identify biomarkers. The clonogenicity and multidifferentiation potential of PDLSCs were identified by colony-forming unit, osteogenic and adipogenic differentiation assays. A CCK8 assay was used to detect the cell activity under different concentrations of mechano-growth factors (MGF-Ct24E). Western blot was used to detect the protein expression of proliferating cell nuclear antigen(PCNA), Scleraxis, collagen type I alpha 1 (COL1A1）, Osterix, Yes-associated protein (YAP) and phosphorylation Yes-associated protein (P-YAP）in PDLSCs. YAP protein expression was observed by immunofluorescence. Knockdown of YAP expression by a siRNA, detected the expression of PCNA, Scleraxis and COL1A1under MGF-Ct24E in PDLSCs.@*Results @#PDLSCs showed high expression of stem cell markers (CD29, CD90 and CD105) and low expression of hematopoietic markers (CD34 and CD45). PDLSCs also have a strong ability to clone. Red calcium junctions were observed by Alizarin red staining, and red lipid droplets were observed by Oil red O staining. After treatment with 50 ng/mL and 100 ng/mL MGF-Ct24E for 24 h, the cell activity of periodontal ligament stem cells was significantly enhanced (P<0.05). The protein expression of PCNA, Scleraxis and COL1A1 was significantly upregulated, and the protein expression of Osterix was significantly downregulated (P<0.05). After treatment with 50 ng/mL mechano-growth factor for 24 h, the phosphorylation of YAP protein was significantly enhanced (P<0.05), and YAP protein was observed to accumulate in the nucleus by immunofluorescence. Following inhibition of YAP expression, PCNA and Scleraxis had significantly downregulated expression caused by MGF-Ct24E (P<0.05). @*Conclusion@#MGF promotes proliferation and fibrogenesis via upregulation of YAP activities.

Autophagy of Vascular Endothelial Cells Influences Diabetic Macroangiopathy： Based on Theory of Qi Deficiency and Stagnation / 中国实验方剂学杂志

The basic pathological change of diabetic macroangiopathy is atherosclerosis （AS）， which is mainly associated with vascular endothelial cells （VECs） injury， oxidative stress， glucose and lipid metabolism disorders， hemorheological abnormalities， and endoplasmic reticulum stress. The injury and dysfunction of VECs are the initiating factors of diabetic macroangiopathy. Autophagy is a subcellular self-protection mechanism that regulates basic intracellular metabolism through lysosome-mediated degradation of proteins and damaged organelles to maintain homeostasis. Insufficient autophagy of VECs leads to enhanced inflammation， apoptosis， and oxidative stress of VECs， which promotes AS. According to the theory of traditional Chinese medicine （TCM）， diabetic macroangiopathy corresponds to the syndrome of internal deficiency and pathogen invasion， with Qi deficiency and stagnation as the key pathogenesis. Qi deficiency is the root cause， and Qi stagnation is the manifestation. The disease occurs with the initial cause of nutrient-defense disharmony and instability of vessels， the main cause of the deficiency of kidney Qi and the lack of source for generation and transformation， the internal cause of Qi and blood loss in the viscera and the stagnation of Qi， blood， and fluid， and the superficial cause of the stagnation of pathological products and the damage of vessels. Autophagy is a microscopic manifestation of Qi， which has the function of dispelling pathogens and maintaining homeostasis. Insufficient autophagy of VECs leads to Qi deficiency and stagnation， and the gradual deficiency and heavy stagnation of Qi lead to insufficient autophagy， which form a vicious cycle. Modern research has demonstrated that regulating the autophagy of VECs is the main way to prevent and treat AS， and TCM can exert the therapeutic effect in a multi-target and multi-pathway manner. Therefore， based on the theory of Qi deficiency and stagnation， the method of tonifying deficiency of and removing stagnation can be adopted to select prescriptions for regulating the autophagy of VECs and treating AS， which can slow down the procession of diabetic macroangiopathy.

Effect of Polydatin on Growth，Apoptosis， and ROS/p38 MAPK Signaling Pathway of Myeloma Cells / 中国实验方剂学杂志

ObjectiveTo analyze the effects of polydatin on myeloma cell growth，apoptosis， and reactive oxygen species（ROS）/p38 mitogen-activated protein kinase（MAPK） signaling pathway. MethodHuman multiple myeloma （MM） cell line U266 cells were cultured in vitro，and the effects of polydatin at 0，20，40，80，160，200 mg·L-1 on the growth of U266 cells were detected by cell counting kit-8（CCK-8）assay. The half-maximal inhibitory concentration（IC50）was calculated. U266 cells in the logarithmic growth phase were randomly divided into a control group， low- and high-dose polydatin （80 and 160 mg·L-1） groups， and a bortezomib （75 nmol·L-1） group. After treatment with corresponding drugs，the cell viability of each group was determined by CCK-8 assay. The apoptosis rate of each group was measured by flow cytometry. The levels of inflammatory factors， such as tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β）， and ROS in each group were measured by enzyme-linked immunosorbent assay （ELISA）. The protein expression levels of apoptosis-related factors， including cysteine aspartate-specific protease-9（Caspase-9），B-cell lymphoma-2-associated X protein（Bax），p38 MAPK，and phosphorylated （p）-p38 MAPK in each group were detected by Western blot. ResultCompared with the results in the control group， polydatin of different concentrations could inhibit the growth of U266 cells （P<0.05），and the effect was potentiated with the increase in the concentration，with IC50 of 156.54 mg·L-1. Compared with the control group，the groups with drug treatment showed blunted cell viability （P<0.05） and increased apoptosis rate，TNF-α，IL-1β，and ROS levels， protein expression levels of Caspase-9， Bax，and p-p38 MAPK/p38 MAPK （P<0.05）. Compared with the low-dose polydatin group， the high-dose polydatin group and the bortezomib group showed improved indicators mentioned above （P<0.05）， and there was no significant difference between the high-dose polydatin group and the bortezomib group. ConclusionPolydatin can activate the ROS/p38 MAPK signaling pathway，promote the expression of inflammatory factors，inhibit the growth of U266 cells，and promote their apoptosis.

Shoutaiwan Ameliorates Oxidative Damage of Human Chorionic Trophoblast Cells by Regulating Nrf2 Signaling Pathway to Treat Recurrent Abortion / 中国实验方剂学杂志

ObjectiveTo study the protective effect of Shoutaiwan-containing serum on the human chorionic trophoblast cells （HTR-8/Svneo） exposed to hydrogen peroxide （H2O2） via the nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway and to decipher the underlying mechanism. MethodThe H2O2 solutions of 25， 50， 100， 200， 400 μmol·L-1 were used to treated the HTR-8/Svneo cells. The cell counting kit-8 （CCK-8） was employed to measure the proliferation of the cells and further determine the optimal concentration of H2O2 solution for modeling and the drug-containing serum. The cells were divided into a blank group， a model group， a dydrogesterone group， and a Shoutaiwan group. The effect of drug-containing serum on H2O2-induced proliferation of HTR-8/Svneo cells was detected by CCK-8 assay. The intracellular reactive oxygen species （ROS） in each group was determined by enzyme-linked immunosorbent assay （ELISA）. Western blot was employed to determine the protein levels of Nrf2， heme oxygenase-1 （HO-1）， quinone oxidoreductase 1 （NQO1）， cysteine-containing aspartate-specific protease-3 （Caspase-3）， and B cell lymphoma/leukemia-2 （Bcl-2）. Cellular immunofluorescence was employed to detect the expression of Nrf2 and Bcl-2. Real-time quantitative polymerase chain reaction （Real-time PCR） was carried out to examine the mRNA level of Nrf2， Caspase-3， and Bcl-2 associated X protein （Bax）. ResultThe optimal concentration of H2O2 for modeling was 50 μmol·L-1， and the optimal concentration of drug-containing serum was 10%. Compared with the blank group， the modeling decreased the proliferation of cells （P<0.01）， increased the intracellular ROS （P<0.01）， down-regulated the protein levels of Nrf2， HO-1， NQO1， and Bcl-2 （P<0.05， P<0.01）， up-regulated the protein levels of Caspase-3 and Bax （P<0.01）， down-regulated the mRNA level of Nrf2 （P<0.01）， and up-regulated the mRNA levels of Caspase-3 and Bax （P<0.05， P<0.01）. Compared with the model group， Shoutaiwan-containing serum increased the proliferation of cells （P<0.01）， reduced the intracellular ROS （P<0.01）， up-regulated the protein levels of Nrf2， HO-1， NQO1， and Bcl-2 （P<0.01）， down-regulated the protein levels of Caspase-3 and Bax （P<0.05， P<0.01）， up-regulated the mRNA level of Nrf2 （P<0.05）， and down-regulated the mRNA levels of Caspase-3 and Bax （P<0.05， P<0.01）. ConclusionShoutaiwan-containing serum can inhibit H2O2-induced apoptosis by activating the Nrf2 signaling pathway and has protective effect on human chorionic trophoblast cells.

Expression and significance of pigment epithelium-derived factor and vascular endothelial growth factor in lens epithelial cells of diabetic patients with cataract / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the expression levels of pigment epithelium-derived factor(PEDF)and vascular endothelial growth factor(VEGF)in lens epithelial cells(LECs)of diabetic patients with age-related cataract, and explore the pathogenesis of diabetic with age-related cataract.METHODS: Retrospectively study. A total of 30 patients with age-related cataract(ARC group)and 30 patients with type 2 diabetes mellitus combined with age-related cataract(DC group)who treated at the Department of Ophthalmology of the First Affiliated Hospital of Bengbu Medical College from August 2020 to April 2021 were collected. Anterior capsule specimens with a diameter of 5.5-6.0mm in the central region of the intraoperative eye were collected during phacoemulsification. The protein expression levels of PEDF and VEGF in LECs were detected by Western-blot. Quantitative real-time PCR(qRT-PCR)was used to detect the relative mRNA expression levels of PEDF and VEGF.RESULTS:PEDF and VEGF were expressed in LECs of both groups, and the relative expression of VEGF mRNA in DC group was 1.364±0.062, which was higher than that of ARC group(1.000±0.0; P&#x003C;0.01). The relative mRNA expression level of PEDF was 0.398±0.053, which was significantly lower than 1.000±0.0 in the ARC group(P&#x003C;0.001). The expression of VEGF protein in LECs was 2.053±0.026 in DC group and 1.680±0.064 in ARC group, respectively. Meanwhile, the expression of PEDF protein was 0.579±0.045 in DC group and 1.058±0.007 in ARC group(all P&#x003C;0.01).CONCLUSION:The changes of PEDF and VEGF expression levels in LECs may be related to the occurrence and development of cataract in diabetic patients.

Progress on the molecular mechanism of action in retinal pigment epithelium during the development of myopia / 国际眼科杂志(Guoji Yanke Zazhi)

Myopia is currently one of the eye diseases that seriously threaten patients' vision worldwide, and its occurrence and development is a complex mechanism. It has been found that retinal pigment epithelium(RPE)cells play a key role in the progression of myopia. RPE cells mainly regulate cell function by regulating the expression of intracellular growth factor and matrix metalloproteinase-2(MMP-2)through the signal pathway of mammalian target of rapamycin(mTOR). At the same time, RPE cells can also be regulated by dopamine receptor agonists, so that cell function changes. When dopamine receptor activation weakened, RPE cell function will be impaired, thus promoting the development of myopia. Studies have shown that the expression of acetylcholine and all-trans retinoic acid in RPE cells can regulate the secretion of growth factors by RPE cells, and the growth factors act on scleral fibroblasts, thus indirectly regulating the course of myopia. Additionally, some studies have shown that RPE cells can coordinate the regulation of γ-aminobutyric acid on scleral cells and indirectly regulate the course of myopia. Besides, the expression of microRNA(microRNA)in RPE cells, such as microRNA-328 and microRNA-29a, was found through previous studies that they can affect the content and composition of extracellular matrix by regulating the expression of MMP-2 in RPE cells, thus leading to the occurrence and development of myopia. Therefore, the expression of multiple signaling pathways and miRNA in RPE cells are closely related to the occurrence and development of myopia. This article reviews the research progress of the molecular mechanism of RPE in the development of myopia, with a view to provide some theoretical basis for the specific molecular mechanism in the development of myopia.

Research progress on the mechanism of ferroptosis in ocular disease associated with retinal pigment epithelial cell injury / 国际眼科杂志(Guoji Yanke Zazhi)

Ferroptosis is a newly identified programmed cell death characterized by lipid peroxidation and iron accumulation. In recent years, with the proposal of the concept of ferroptosis as well as the deepening of its mechanism research, great breakthroughs have been made in the exploration of the pathogenesis of ocular diseases related to the function decline of retinal pigment epithelium cells(RPECs), such as age-related macular degeneration(AMD), retinitis pigmentosa(RP)and diabetic retinopathy(DR). This article reviews the basic concept of ferroptosis, the main mechanism of ferroptosis in RPECs and the role of ferroptosis regulation in the development of RPECs-related ocular diseases, hoping to provide references for the study of the pathogenesis of RPECs-related ocular diseases.

Advances in neuroprotection of Sigma-1 receptor in retinal ganglion cell / 国际眼科杂志(Guoji Yanke Zazhi)

With complicated pathogenesis and limited treatment options, optic neuropathy is one of the major blinding diseases characterized by damaged retinal ganglion cells(RGC)and axons. Sigma-1 receptor(S1R)is a chaperone protein mainly located in the endoplasmic reticulum membrane, which is abundant in retina and highly expressed in the ganglion cell layer. S1R has been noted increasingly as a novel target for the treatment of neuro-degenerations. More studies have shown that S1R is a pluripotent modulator including Ca2+ homeostasis, endoplasmic reticulum stress response, oxidative stress response, neurotrophic factor secretion and glial cell activation, indicating that S1R can generate significant impacts on neuroprotection in neurodegenerative diseases. Additionally, S1R also has neuroprotective effects against RGC loss and dysfunction both in vivo and in vitro, reversing loss partially and maintaining structural integrity, while the absence of S1R exacerbates the disease or increases vulnerability to degenerative diseases. This article intends to review the progress and mechanisms in neuroprotection of S1R in RGC, aiming to provide a new target for the treatment of optic neuropathy.

Protective effect of Modified Taohong Siwu Decoction on hypoxia injury to retinal Müller cells / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the protective effect of Modified Taohong Siwu Decoction(MTSD)on hypoxia injury to retinal Müller cells rMC-1.METHODS: Retinal Müller cells rMC-1 were interfered with the MTSD drug-containing serum under hypoxia condition, and were randomly divided into control group(21%O2), hypoxia model group(1%O2), MTSD drug-containing serum low-dose(1%O2+5% medicated serum), medium-dose(1%O2+10% medicated serum)and high-dose(1%O2+15%medicated serum)groups. Cell viability was detected by CCK-8 method, and secretion of vascular endothelial growth factor(VEGF)and pigment epithelium derived factor(PEDF)was detected by ELISA. The protein expressions of p-STAT3, STAT3 and hypoxia-inducible factor-1α(HIF-1α)were detected by Western blot, and the gene expressions of VEGF, PEDF, STAT3 and HIF-1α were detected by real-time polymerase chain reaction(PCR).RESULTS: The viability of rMC-1 cells was significantly inhibited when cultured at 1%O2 for 48h compared with that of control group(P&#x003C;0.05), while it was improved in both low and medium dose of MTSD groups(P&#x003C;0.05). The viability of rMC-1 cells in high dose group was not improved in hypoxia condition(P&#x003E;0.05). The low and medium doses of MTSD could reduce the protein expressions of VEGF in supernatant of rMC-1 cells under hypoxia condition(P&#x003C;0.05), while the protein expressions of PEDF could not be increased(P&#x003E;0.05). The above two dose groups down-regulated the protein levels of both p-STAT3 and HIF-1α(P&#x003C;0.05), and the inhibition effect of low dose group was better than that of medium dose group(P&#x003C;0.05). The medium dose of MTSD could up-regulate STAT3 protein level after hypoxia culture in rMC-1 cells(P&#x003C;0.05). The low and medium doses of MTSD significantly down-regulated VEGF gene level(P&#x003C;0.05)and up-regulated PEDF gene level after hypoxia culture in rMC-1 cells(P&#x003C;0.05), and the function in the low dose group was superior to that in the medium dose group(P&#x003C;0.05). The low dose of MTSD could down-regulate STAT3 and HIF-1α gene levels after hypoxia culture in rMC-1 cells(P&#x003C;0.05).CONCLUSION: Probably by inhibiting the STAT3/HIF-1α pathway, the drug-containing serum of MDST down-regulated the expression of VEGF protein and gene in hypoxia-induced retinal Müller cells, rMC-1, up-regulated the gene expression of PEDF, and alleviated the hypoxia injury to the cells.

Synergistic effect of sorafenib with Platycladus orientalis (L) leaf extract on cervical cancer

The extracts of Platycladus orientalis (L.) Franco leaves have shown promising anti-cancer, anti-oxidant and anti-inflammatory potency with the traditional knowledge of healing HPV associated warts. The purpose of this research is to assess the synergistic activity of sorafenib and Platycladus orientalis (L) leaf extraction on cervical cancer cells. The cytotoxicity efficiency of different concentrations of Sorafenib and ethanol extract of Platycladus orientalis (L.) leaves were tested on HeLa cells by MTT and Trypan blue assays. The synergistic effect of the IC50 concentrations of Sorafenib and Platycladus orientalis (L.) on HeLa cell by MTT assay, and mRNA expression levels of tumor suppressor tazarotene-induced gene 3 (TIG3), proliferating cell nuclear antigen (PCNA) gene and apoptosis modulator (Bcl-2) gene by RT-PCR were evaluated with individual treatments. Combination treatment showed a relatively more expression of TIG3 and less expression of Bcl-2 and PCNA was observed. Growth factor-induced MAPKP activation was arrested by compound combination treatment, which and suppression of proliferation-induced apoptosis of cervical cancer cells. Based on the our results, the combination of sorafenib and crude leaf extract from Platycladus orientalis (L.) can effectively suppress cervical cancer cell growth, thereby providing an interesting rationale for further clinical trials and in-vivo studies.

Anatomy and histology of the digestive tract and immunolocalization of Npy in the fish Cichlasoma dimerus (Cichliformes: Cichlidae) / Anatomía e histología del tracto digestivo e inmunolocalización del Npy (péptido orexigénico) en el pez Cichlasoma dimerus (Cichliformes: Cichlidae)

Abstract Introduction: The digestive tract of Neotropical cichlids has not been deeply studied, and it is a fundamental topic for understanding fish physiology, nutrition, trophic associations, and evolution. Objective: To describe anatomically and histologically the digestive tract of the Neotropical cichlid fish Cichlasoma dimerus and to immunolocalize the orexigenic peptide (Npy) along the intestine. Methods: We euthanized 14 adult individuals and fixed the organs in Bouin´s solution; we stained 7 μm thick paraffin sections for general description and with Alcian Blue (pH = 2.5, AB) and Periodic acid-Schiff (PAS) to identify acid or neutral glycoconjugates, respectively. Additionally, we performed immunohistochemistry for Npy in 3 adult individuals. We manually counted PAS- and AB-positive cells, and Npy-immunoreactive cells per fold. Results: There is a short oesophagus, a sac-like stomach, and a tubular intestine with two loops. The oesophagus has a stratified epithelium with a high density of PAS- and AB-positive goblet cells and striated muscle fibers in the tunica muscularis. The stomach mucosa is formed by simple columnar epithelium. The intestine has a simple columnar epithelium, with brush border and interspersed PAS- and AB-positive goblet cells, and Npy-immunoreactive cells. There is an ileorectal valve in the transition between the posterior intestine and the rectum. This last gut portion has goblet cells and a thicker tunica muscularis. Conclusions: C. dimerus shares features with other Neotropical cichlids, but the goblet cells and gastric glands distribution seems to be unique for the species. To our understanding, this is the first work to describe Npy-immunoreactive cells distribution in the intestine of a Neotropical cichlid fish.

Resumen Introducción: El tracto digestivo de los cíclidos neotropicales no ha sido profundamente estudiado y es un tema fundamental para entender la fisiología, nutrición, asociaciones tróficas y evolución de los peces. Objetivo: Describir anatómica e histológicamente el tracto digestivo del pez cíclido neotropical Cichlasoma dimerus e inmunolocalizar el péptido orexigénico (Npy) a lo largo del intestino. Métodos: Sacrificamos 14 individuos adultos y fijamos los órganos en solución de Bouin; teñimos secciones de parafina de 7 μm de espesor para una descripción general y con azul alcián (pH = 2.5, AB) y ácido periódico-Schiff (PAS) para identificar glicoconjugados ácidos o neutros, respectivamente. Además, en 3 individuos adultos se realizaron inmunohistoquímicas contra Npy. Contamos manualmente las células PAS y AB positivas, y las células inmunorreactivas a Npy por pliegue. Resultados: Hay un esófago corto, un estómago en forma de saco y un intestino con dos vueltas. El esófago tiene un epitelio estratificado con una alta densidad de células caliciformes PAS- y AB- positivas y fibras esqueléticas estriadas en las capas musculares. La mucosa del estómago está revestida por epitelio simple cilíndrico. El epitelio intestinal es simple cilíndrico con chapa estriada y células caliciformes PAS- y AB- positivas intercaladas, y células inmunorreactivas a Npy. Hay una válvula ileorrectal en la transición entre el intestino posterior y el recto. Esta última porción intestinal tiene células caliciformes y una túnica muscular más gruesa. Conclusiones: C. dimerus comparte características con otros cíclidos neotropicales, pero la distribución de las células caliciformes y las glándulas gástricas, serían rasgos propios de esta especie. A nuestro entender, este es el primer trabajo que describe la distribución de células inmunorreactivas a Npy en el intestino de un pez cíclido neotropical.

Sinovite vilonodular pigmentada da articulação temporomandibular: uma revisão integrativa / Pigmented villonodular synovitis of the temporomandibular joint: an integrative review / Sinovitis villonodular pigmentada de la articulación temporomandibular: una revisión integradora

Objetivo: Avaliar os casos nos quais os pacientes apresentassem SVNP na ATM, incluindo aspectos clínicos, imaginológicos, histopatológicos e tratamento. Metodologia: Trata-se de uma revisão integrativa com dados obtidos nas bases de dados SciELO, PubMed, Medline e Lillacs entre 1982 e 2021, através dos descritores: "Case report", "Temporomandibular Joint", "Pigmented Villonodular Synovitis". Critérios de inclusão: estudos de relato de caso, textos completos disponíveis, idiomas de publicação em inglês, português ou espanhol. Critérios de exclusão: estudos sem presença de aspectos clínicos, relatos não localizados na ATM, artigos de metanálise, revisão sistemática e de literatura, ensaios clínicos, capítulos de livro, dissertações e teses. Dos 156 resultados, apenas 23 compuseram a revisão. Resultados: Como tratamento, a ressecção total através da cirurgia aberta é recomendada. Os sintomas mais comuns foram: dor na mastigação, trismo, dor pré auricular, dormência, parestesia, perca auditiva e inchaço da glândula parótida. Conclusão: Os aspectos imaginológicos revelam erosão de fossa glenóide e côndilo, histopatologicamente, células gigantes com depósito de hemossiderina, e o tratamento recomendado, ressecção via cirurgia aberta com posterior curetagem... (AU)

Objective: To evaluate the cases in which patients presented PVNS in the TMJ, including clinical, imaging, histopathological and treatment aspects. Methodology: This is an integrative review with data obtained from the SciELO, PubMed, Medline and Lillacs databases between 1982 and 2021, using the descriptors: "Case report", "Temporomandibular Joint", "Pigmented Villonodular Synovitis". Inclusion criteria: case report studies, full texts available, languages of publication in English, Portuguese or Spanish. Exclusion criteria: studies without the presence of clinical aspects, reports not located in the TMJ, meta analysis articles, systematic and literature reviews, clinical trials, book chapters, dissertations and theses. Of the 156 results, only 23 made up the review. Results: As a treatment, total resection through open surgery is recommended. The most common symptoms were: chewing pain, trismus, pre-auricular pain, numbness, paresthesia, hearing loss and parotid gland swelling. Conclusion: The imaging findings reveal erosion of the glenoid fossa and condyle, histopathologically, giant cells with hemosiderin deposits, and the recommended treatment, resection via open surgery with subsequent curettage... (AU)

Objetivo: Evaluar los casos en que los pacientes presentaron SVNP en la ATM, incluyendo aspectos clínicos, imagenológicos, histopatológicos y tratamiento. Metodología: Se trata de una revisión integradora con datos obtenidos de las bases de datos SciELO, PubMed, Medline y Lillacs entre 1982 y 2021, utilizando los descriptores: "Caso clínico", "Articulación temporomandibular", "Sinovitis villonodular pigmentada". Criterios de inclusión: estudios de casos clínicos, textos completos disponibles, idiomas de publicación en inglés, portugués o español. Criterios de exclusión: estudios sin aspectos clínicos, informes no localizados en la ATM, artículos de metanálisis, revisiones sistemáticas y de literatura, ensayos clínicos, capítulos de libros, disertaciones y tesis. De los 156 resultados, 23 conformaron la revisión. Resultados: Como tratamiento se recomienda la resección total mediante cirugía abierta. Los síntomas más frecuentes fueron: dolor masticatorio, trismus, dolor preauricular, entumecimiento, parestesia, hipoacusia e inflamación de glándula parótida. Conclusión: Los hallazgos imagenológicos revelan erosión de fosa glenoidea y cóndilo, histopatológicamente células gigantes con depósitos de hemosiderina y el tratamiento recomendado, resección abierta con posterior curetaje... (AU)

Effect of crosstalk between Th17 and Th9 cells on the activation of dermal vascular smooth muscle cells in systemic scleroderma and regulation of tanshinone IIA

Abstract Background: To evaluate the effect of T-helper 17 (Th17) cells and Th9 cells on the activation of dermal vascular smooth muscle cells (DVSMCs) in systemic scleroderma (SSc) and regulation of tanshinone IIA. Methods: The expression of interleukin 17 receptor (IL-17R) and interleukin 9 receptor (IL-9R) in the skin of SSc patients was assessed by immunofluorescence. The expression of IL-9 and IL-9R mRNA in peripheral blood mononuclear cells (PBMCs) of SSc patients were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The proportion of Th9 cells in PBMCs of SSc patients was sorted by flow cytometry. The effect of IL-9 on the differentiation of Th17 and IL-17 on that of Th9 was detected by flow cytometry. The proportion of Th9 and Th17 cells in SSc patients was detected by flow cytometry. The level of collagen I, III, α-SMA, IL-9R, IL-17R, JNK, P38, and ERK were analyzed using western blot (WB). Results: Th9 cells were highly expressed in SSc. IL-9 stimulated the differentiation of immature T cells into Th17 cells. IL-17 induced the differentiation of immature T cells intoTh9 cells. Tanshinone IIA inhibited the differentiation of immature T lymphocytes into Th17 and Th9. WB showed that the combined action of IL-17 and IL-9 upregulated the inflammation and proliferation of DVSMCs. Anti-IL17, anti-IL9, and tanshinone IIA inhibited the functional activation of DVSMCs. Study limitations: For Th17, Th9 and vascular smooth muscle cells, the study on the signal pathway of their interaction is not thorough enough. More detailed studies are needed to explore the mechanism of cell-cell interaction. Conclusions: The current results suggested that Th17 and Th9 cells induced the activation of DVSMCs in SSc through crosstalk in vitro, and tanshinone IIA inhibited the process.

Luschka and the carotid body / Luschka e o corpo carotídeo

The 'carotid body' is a small structure sited at the bifurcation of the common carotid artery. The macroscopic features of the carotid body, and items of the extrinsic nervous, and vascular supplies, were initially described by Taube (1743), complemented by a number of authors that followed, proceeding until Luschka (1962), who added the first microscopic study. The macroscopic features of the carotid body, including location, extrinsic innervation, and vascular supply, then provided, were described in a relatively satisfactory manner. However, despite Luschka's great and admirable effort, the microscopic findings seem to be flawed, what can be ascribed to the technical limitations at the time, and the artifacts due to the used procedures. Nevertheless, there is no doubt that Luschka and his forerunners provided an important step for forthcoming research on the carotid body, and its innervation.

O 'corpo carotídeo' é uma pequena estrutura situada na bifurcação da artéria carótida comum. Os aspectos macroscópicos do corpo carotídeo e itens sobre o suprimento nervoso e vascular extrínsecos foram descritos inicialmente por Taube (1743), complementados por um certo número de autores que seguiram, prosseguindo até Luschka (1962), que acrescentou o primeiro estudo microscópico. Os aspectos macroscópicos do corpo carotídeo, incluindo localização, inervação extrínseca e suprimento vascular, então providos, foram descritos de modo relativamente satisfatório. Entretanto, apesar do grande e admirável esforço de Luschka, os achados microscópicos aparecem falhos, o que pode ser atribuído às limitações técnicas daquele tempo e a artefatos devidos aos procedimentos utilizados. Todavia, não há dúvida que Luschka e seus precursores proveram um importante passo para pesquisas que vieram sobre o corpo carotídeo e da sua inervação.