Upregulation of PGC-1α expression by pioglitazone mediates prevention of sepsis-induced acute lung injury

Abstract The imbalance between pro-inflammatory M1 and anti-inflammatory M2 macrophages plays a critical role in the pathogenesis of sepsis-induced acute lung injury (ALI). Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) may modulate macrophage polarization toward the M2 phenotype by altering mitochondrial activity. This study aimed to investigate the role of the PGC-1α agonist pioglitazone (PGZ) in modulating sepsis-induced ALI. A mouse model of sepsis-induced ALI was established using cecal ligation and puncture (CLP). An in vitro model was created by stimulating MH-S cells with lipopolysaccharide (LPS). qRT-PCR was used to measure mRNA levels of M1 markers iNOS and MHC-II and M2 markers Arg1 and CD206 to evaluate macrophage polarization. Western blotting detected expression of peroxisome proliferator-activated receptor gamma (PPARγ) PGC-1α, and mitochondrial biogenesis proteins NRF1, NRF2, and mtTFA. To assess mitochondrial content and function, reactive oxygen species levels were detected by dihydroethidium staining, and mitochondrial DNA copy number was measured by qRT-PCR. In the CLP-induced ALI mouse model, lung tissues exhibited reduced PGC-1α expression. PGZ treatment rescued PGC-1α expression and alleviated lung injury, as evidenced by decreased lung wet-to-dry weight ratio, pro-inflammatory cytokine secretion (tumor necrosis factor-α, interleukin-1β, interleukin-6), and enhanced M2 macrophage polarization. Mechanistic investigations revealed that PGZ activated the PPARγ/PGC-1α/mitochondrial protection pathway to prevent sepsis-induced ALI by inhibiting M1 macrophage polarization. These results may provide new insights and evidence for developing PGZ as a potential ALI therapy.

Higher-order aberrations and visual quality after incision lenticule extraction surgery with intraoperative angle kappa adjustments between small and large kappa patients: A 2-year follow-up

Purpose: To evaluate the postoperative visual outcomes, that is, corneal higher?order aberrations (HOAs) and visual quality, of patients with an angle kappa greater than 0.30 mm who underwent angle kappa adjustment during small?incision lenticule extraction (SMILE) 2 years after surgery compared to eyes with an angle kappa less than 0.30 mm. Methods: This was a retrospective study and included 12 patients from October 2019 to December 2019 who underwent the SMILE procedure for correction of myopia and myopic astigmatism and had one eye with a large kappa angle and another eye with a small kappa angle. Twenty?four months after surgery, an optical quality analysis system (OQAS II; Visiometrics, Terrassa, Spain) was used to measure the modulation transfer function cutoff frequency (MTFcutoff), Strehl2D ratio, and objective scatter index (OSI). HOAs were measured with a Tracey iTrace Visual Function Analyzer (Tracey version 6.1.0; Tracey Technologies, Houston, TX, USA). Assessment of subjective visual quality was achieved using the quality of vision (QOV) questionnaire. Results: At 24 months postoperatively, the mean spherical equivalent (SE) refraction was ? 0.32 ± 0.40 and ? 0.31 ± 0.35 in the S?kappa group (kappa <0.3 mm) and the L?kappa group (kappa ?0.3 mm), respectively (P > 0.05). The mean OSI was 0.73 ± 0.32 and 0.81 ± 0.47, respectively (P > 0.05). There was no significant difference in MTFcutoff and Strehl2D ratio between the two groups (P > 0.05). Total HOA, coma, spherical, trefoil, and secondary astigmatism were not significantly different (P > 0.05) between the two groups. Conclusion: Adjustment of angle kappa during SMILE helps reduce the decentration, results in less HOAs, and promotes visual quality. It provides a reliable method to optimize the treatment concentration in SMILE.

Research progress on the role of environmentally induced DNA methylation in congenital cleft lip and/or palate / 口腔疾病防治

@#Congenital cleft lip and/or palate (CL/P) is a common malformation of maxillofacial development. At present, it is believed that the etiology of congenital cleft lip and palate mainly results from genetic factors and environmental factors. Epigenetic changes induced by environmental factors may be the key factor in the occurrence of fetal congenital malformations. As one of the important epigenetic modifications, DNA methylation has been widely and deeply studied in many fields, but as a link between the individual and the environment, its application in CL/P is limited. Existing studies have shown that DNA methylation is closely related to the occurrence of cleft lip and palate. Stimulation of folate deficiency, smoking, pollutant exposure and other environmental factors can induce changes in the state of DNA methylation, thus affecting gene expression in the development of lip and palate and leading to the occurrence of deformities.

Propofol inhibits myocardial injury induced by microvesicles derived from hypoxiareoxygenated endothelial cells via lncCCT4-2/CCT4 signaling

BACKGROUND: Ischemia-reperfusion (IR) induces increased release of extracellular vesicles in the heart and exacerbates myocardial IR injury. We have previously shown that propofol attenuates hypoxia/reoxygenation (HR)-induced injury in human umbilical vein endothelial cells (HUVECs) and that microvesicles derived from propofol-treated HUVECs inhibit oxidative stress in endothelial cells. However, the role of microvesicles derived from propofol post-treated HUVECs ((HR + P)-EMVs) in IR-injured cardiomyocytes is unclear. In this study, we aimed to investigate the role of (HR + P)-EMVs in cardiac IR injury compared to microvesicles derived from hypoxic/reoxygenated HUVECs (HR-EMVs) and to elucidate the underlying mechanisms. METHODS: Hypoxia/reoxygenation (HR) models of HUVECs and AC16 cells and a mouse cardiac IR model were established. Microvesicles from HR-injured HUVECs, DMSO post-treated HUVECs and propofol post-treated HUVECs were extracted by ultra-high speed centrifugation, respectively. The above EMVs were co-cultured with HR-injured AC16 cells or injected intracardially into IR mice. Flow cytometry and immunofluorescence were used to determine the levels of oxidative stress and apoptosis in cardiomyocytes. Apoptosis related proteins were detected by Western blot. Echocardiography for cardiac function and Evans blue-TTC staining for myocardial infarct size. Expression of lncCCT4-2 in EMVs and AC16 cells was analysed by whole transcriptome sequencing of EMVs and RT-qPCR. The molecular mechanism of inhibition of myocardial injury by (HR + P)-EMVs was elucidated by lentiviral knockdown of lncCCT4-2, plasmid overexpression or knockdown of CCT4, and actinomycin D assay. RESULTS: In vitro and in vivo experiments confirmed that HR-EMVs exacerbated oxidative stress and apoptosis in IR-injured cardiomyocytes, leading to increased infarct size and worsened cardiac function. Notably, (HR + P)-EMVs induced significantly less oxidative stress and apoptosis in IR-injured cardiomyocytes compared to HR-EMVs. Mechanistically, RNA sequencing of EMVs and RT-qPCR showed that lncCCT4-2 was significantly upregulated in (HR + P)-EMVs and cardiomyocytes co-cultured with (HR + P)-EMVs. Reduction of lncCCT4-2 in (HR + P)-EMVs enhanced oxidative stress and apoptosis in IR-injured cardiomyocytes. Furthermore, the anti-apoptotic activity of lncCCT4-2 from (HR + P)-EMVs was achieved by increasing the stability of CCT4 mRNA and promoting the expression of CCT4 protein in cardiomyocytes. CONCLUSIONS: Our study showed that (HR + P)-EMVs uptake by IR-injured cardiomyocytes upregulated lncCCT4-2 in cardiomyocytes and promoted CCT4 expression, thereby inhibiting HR-EMVs induced oxidative stress and apoptosis. Highlights Microvesicles from hypoxic/reoxygenated HUVECs (HR-EMVs) exacerbated oxidative stress and apoptosis in IR-injured cardiomyocytes. Microvesicles from propofol post-treated HUVECs ((HR + P)-EMVs) induced diminished oxidative stress and apoptosis in IR-injured cardiomyocytes compared with microvesicles from hypoxic/reoxygenated HUVECs (HR-EMVs). lncCCT4-2 was significantly highly expressed in (HR + P)-EMVs and cardiomyocytes co-cultured with (HR + P)-EMVs, and reduction of lncCCT4-2 in (HR + P)-EMVs enhanced oxidative stress and apoptosis in IR-injured cardiomyocytes. lncCCT4-2 inhibited HR-EMVs induced oxidative stress and apoptosis in HR-injured AC16 cells by increasing the stability of CCT4 mRNA and promoting the expression of CCT4 protein in AC16 cells.

The clinical treatment of sports to relieve the difficulty of falling asleep / O tratamento clínico de esportes para aliviar a dificuldade em adormecer / O tratamiento clínico de deportes para aliviar la dificultad en dormirse

ABSTRACT Introduction: Difficulty falling asleep is connected to the malfunctioning of the sleep and wakefulness mechanism of the human body caused by various reasons. There are a series of adverse reactions resulting from abnormal or poor quality of sleep during sleep per se. This symptom severely affects an individual's physical condition and mental health. Objective: To explore the effect of physical exercise on patients with difficulty falling asleep. Methods: Mathematical statistics were used to analyze 60 patients with difficulty falling asleep. We divided the patients into a sports group and a control group. The patients in the sports group took sports training, while the control group did not. After two weeks of intervention and comparison, we used mathematical statistics to evaluate the groups' cognitive function. Results: After physical exercise, the patient's sleep quality was higher and sleep disorders were reduced. Conclusion: Physical activity is a simple and popular way of exercising. It is effective in improving the cognitive function of patients with difficulty falling asleep. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: A dificuldade em adormecer está ligada ao mau funcionamento do mecanismo do ciclo sono-vigília no corpo humano e é causada por diversos motivos. Existe uma serie de reações adversas que resultam da má qualidade - ou qualidade anormal - do sono, especificamente durante o sono. Esses sintomas afetam seriamente a condição física e a saúde mental das pessoas. Objetivo: Explorar o efeito de exercícios físicos em pacientes com dificuldade em adormecer. Métodos: Estatísticas matemáticas foram usadas para analisar 60 pacientes com dificuldade em adormecer. Dividimos os pacientes em um grupo de esportes e outro de controle. Os pacientes no grupo de esportes tiveram treinamento esportivo, enquanto o grupo de controle não. Após duas semanas de intervenção e comparação, usamos estatísticas matemáticas para avaliar a função cognitiva dos grupos. Resultados: Após o exercício físico, houve melhora na qualidade de sono dos pacientes e os transtornos do sono diminuíram. Conclusão: A atividade física é uma forma simples e popular de se exercitar. É eficaz na melhora da função cognitiva de pacientes com dificuldade de adormecer. Nível de evidência II; Estudos terapêuticos - investigação de resultados de tratamento.

RESUMEN Introducción: La dificultad en dormirse se relaciona al mal funcionamiento del mecanismo del ciclo sueño-vigilia en el cuerpo humano y es causada por diversos motivos. Existe una serie de reacciones adversas que resultan de la mala calidad o calidad anormal del sueño, durante el sueño en sí. Esos síntomas afectan seriamente la condición física y la salud mental de las personas. Objetivo: Explorar el efecto de ejercicios físicos en pacientes con dificultad en dormirse. Métodos: Dividimos los pacientes en un grupo de deportes y otro de control. Los pacientes en el grupo de deportes tuvieron entrenamiento deportivo, mientras el grupo de control no. Tras dos semanas de intervención y comparación, usamos estadísticas matemáticas para evaluar la función cognitiva de los grupos. Resultados: Tras el ejercicio físico, hubo mejoría en la calidad del sueño de los pacientes y los trastornos del sueño disminuyeron. Conclusión: La actividad física es una forma simple y popular de ejercitarse. Es eficaz en la mejoría de la función cognitiva de pacientes con dificultad de dormirse. Nivel de evidencia II; Estudios terapéuticos - investigación de resultados de tratamiento.

Investigation on the influence of periodontal disease in gestation on small for gestational age / 口腔疾病防治

Objective @#The purpose of this study was to investigate the relevant social and environmental factors affecting the occurrence of periodontal diseases during pregnancy in pregnant women and to analyze the influence of the periodontal status of women in the second trimester of pregnancy on small for gestational age (SGA) delivery.@*Methods@# A total of 215 pregnant women were enrolled in this study in the Department of Periodontology of the West China Hospital of Stomatology of Sichuan University from May 2015 to May 2018. Periodontal parameters, such as bleeding on probing (BOP), probing depth (PD) and clinical attachment loss (CAL), were recorded at 16-24 weeks of gestational age. Subjects were divided into the periodontitis (n=32) group, gingivitis (n=171) group and periodontally healthy (n=12) group according to their periodontal conditions. With the patient′s informed consent, the patient decided whether to receive periodontal treatment. Basic and socioeconomic information was collected through questionnaires. After delivery, subjects were divided into the SGA group and non-SGA group according to their birth results. The periodontal clinical indicators, questionnaire results and delivery results were compared among the groups.@*Results @#The mean PD (P=0.005, r=-0.192) and BOP% (P=0.003, r=-0.199) were negatively correlated with economic income. The family income in the periodontitis group was significantly lower than that in the healthy group and the gingivitis group (P < 0.05). The flossing use rate was significantly higher in the healthy group than that in the gingivitis group (P < 0.05). A total of 106 pregnant women received scaling and root planing, while 109 patients only received oral hygiene instruction. After delivery, SGA occurred in 23 cases (10.7%), and there were no significant difference in SGA incidence among the three groups (P > 0.05). PD ≥ 5 mm% and PD ≥ 4 mm% (P < 0.05) were significantly higher in the SGA group than in the non-SGA group. There was no significant difference in SGA incidence between the treated group and the untreated group (P > 0.05).@*Conclusion@#Family income and dental flossing use have an impact on the incidence of periodontal diseases during pregnancy. The severity of periodontitis in pregnant women is correlated with the incidence of SGA.

Effect of lncRNA CCAT2 on proliferation and cell cycle of cervical cancer CaSki cells / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of long non-coding RNA (lncRNA)-CCAT2 on the proliferation and cell cycle of cervical cancer cells. Methods: The expression of CCAT2 in 3 cervical cancer cell lines (HeLa, C-33A, and CaSki) was detected by qPCR and the cell line with the highest expression level was selected for subsequent experiments. CCAT2 overexpression and interference vectors were designed and synthesized. After transfection, qPCR was performed to detect the transfection efficiency. The cells were divided into 5 groups: control, sh-EV (empty vector), overExp-EV , sh-CCAT2, and overExp-CCAT2. MTT assay was performed to evaluate cell viability. Flow cytometry was performed to measure cell cycle. WB was performed to detect the expressions of Ki67, cyclin D1, and cyclin dependent kinase 4 (CDK4). Results: Among HeLa, C-33A, and CaSki cells, the highest expression of CCAT2 was found in CaSki cells. CCAT2 overexpression and interference vectors were successfully transfected into the CaSki cells. Compared with the control group, the cells viability and proliferation in the sh-CCAT2 group was significantly decreased (all P<0.01), the proportion of cells in the G1 phase was significantly increased (P<0.01), and the expression levels of Ki67, cyclin D1, and CDK4 were significantly decreased (all P<0.01). However, in the overExp-CCAT2 group, the cell proliferation was enhanced and the expression levels of Ki67, cyclin D1, and CDK4 were significantly increased (all P<0.01). Conclusion: CCAT2 affects proliferation and cell cycle of cervical cancer cells by regulating the expressions of their associated proteins.

A comparison of the sealing abilities between Biodentine and MTA as root-end filling materials and their effects on bone healing in dogs after periradicular surgery

ABSTRACT Objectives: To compare the sealing ability and biocompatibility of Biodentine with mineral trioxide aggregate (MTA) when used as root-end filling materials. Methodology: The Cell Counting Kit-8 (CCK-8) assay was used to compare the cytotoxicity of MTA and Biodentine. Twenty-one extracted teeth with a single canal were immersed in an acidic silver nitrate solution after root-end filling. Then, the volume and depth of silver nitrate that infiltrated the apical portion of the teeth were analyzed using micro-computed tomography (micro-CT). Seventy-two roots from 3 female beagle dogs were randomly distributed into 3 groups and apical surgery was performed. After six months, the volume of the bone defect surrounding these roots was analyzed using micro-CT. Results: Based on the results of the CCK-8 assay, MTA and Biodentine did not show statistically significant differences in cytotoxicity (P>0.05). The volume and the depth of the infiltrated nitrate solution were greater in the MTA group than in the Biodentine group (P<0.05). The volume of the bone defect was larger in the MTA group than in the Biodentine group. However, the difference was not significant (P>0.05). The volumes of the bone defects in the MTA and Biodentine groups were smaller than the group without any filling materials (P<0.05). Conclusions: MTA and Biodentine exhibited comparable cellular biocompatibility. Biodentine showed a superior sealing ability to MTA in root-end filling. Both Biodentine and MTA promoted periradicular bone healing in beagle dog periradicular surgery models.

Expression, purification and polyclonal antibody preparation of GST-Ccd1 fusion protein / 中国应用生理学杂志

<p><b>AIM</b>By using of Escherichia coli DH5alpha to express GST-Ccd1 fusion protein and which was purified by affinity chromatographic separation. The purified target protein was used to immunize rabbits to prepare polyclonal antibody.</p><p><b>METHODS</b>The previously constructed recombinant prokaryotic expression vector pGEX-5X-1-Ccd1 was transformed into Escherichia coli DH5alpha and was induced to expression by IPTG. The recombinant target protein was expressed with soluble state in Escherichia coli expression system which was separated and purified by chromatographic column stuffed with glutathione Sepharose 4B. The prepared antigen was used to immunize rabbits to get anti-Ccd1 specific rabbit original polyclonal antibody.</p><p><b>RESULTS</b>ELISA data demonstrated that the antibody titer of the serum was up to 1:40 000. Immunohistochemistry analysis indicated that the "home-made" antibody had a specific interaction with Ccd1 protein and which could be used for extended experimental research.</p><p><b>CONCLUSION</b>The anti-Ccd1 polyclonal antibody we had prepared had a high quality of potency and specificity. The antibody was demonstrated by experiments that which could totally fulfill the requirement of immunoblotting and immunohistochemistry study of Ccd1. The antibody provided an useful experimental tool to profoundly research the tissue expression profile, intercellular location and biological function of Ccd1.</p>