Inhibition of Proliferation, Migration, and Invasion of Keloid Fibroblasts By miR-183-5p Through Downregulating EGR1 / Inhibición de la Proliferación, Migración e Invasión de Fibroblastos Queloides por miR-183-5p Mediante la Regulación Negativa de EGR1

SUMMARY: Keloid scar is a unique benign fibroproliferative tumor of the human skin. Previously, it was reported that early growth response 1 (EGR1), a transcription factor, promotes keloid fibrosis; however, the mechanism by which EGR1 modulates keloid formation was not elaborated. In this research, the specific function and the microRNA (miRNA) regulatory network of EGR1 in keloids was examined. Keloid fibroblasts (KFs) were transfected with EGR1-small interfering RNA (siEGR1), EGR1-overexpression plasmid (pcDNA3.1-EGR1), and microRNA (miR-183-5p)-mimics to regulate the expression of EGR1 and miR-183-5p. The study employed dual-luciferase reporter assays to explore the targeting regulation of miR-183-5p on EGR1. Additionally, Western blotting, flow cytometry, qRT-PCR, cell count kit-8 (CCK-8), transwell, and wound healing assays, and RNA sequencing were conducted. EGR1 was upregulated in KFs, and EGR1 silencing diminished proliferation, fibrosis, migration, invasion, and apoptosis of cells. In KFs, the expression of miR- 183-5p was reduced, leading to the inhibition of cell proliferation, migration, and invasion. Conversely, it enhanced apoptosis. By targeting EGR1, miR-183-5p partially counteracted the impact of EGR1 on migration, invasion, and fibrosis in KFs. The findings imply that miR-183-5p suppresses keloid formation by targeting EGR1. As a result, EGR1 holds promise as a potential therapeutic target for preventing and treating keloids.

La cicatriz queloide es un tumor fibroproliferativo benigno único de la piel humana. Anteriormente, se informó que la respuesta de crecimiento temprano 1 (EGR1), un factor de transcripción, promueve la fibrosis queloide; sin embargo, no se explicó el mecanismo por el cual EGR1 modula la formación de queloides. En esta investigación, se examinó la función específica y la red reguladora de microARN (miARN) de EGR1 en queloides. Se transfectaron fibroblastos queloides (KF) con ARN de interferencia pequeño de EGR1 (siEGR1), plásmido de sobreexpresión de EGR1 (pcDNA3.1-EGR1) y miméticos de microARN (miR-183-5p) para regular la expresión de EGR1 y miR-183. -5p. El estudio empleó ensayos de indicador de luciferasa dual para explorar la regulación dirigida de miR-183-5p en EGR1. Además, se realizaron pruebas de transferencia Western, citometría de flujo, qRT-PCR, kit de recuento celular-8 (CCK-8), transwell y curación de heridas, y secuenciación de ARN. EGR1 estaba regulado positivamente en KF, y el silenciamiento de EGR1 disminuyó la proliferación, fibrosis, migración, invasión y apoptosis de las células. En KF, la expresión de miR- 183-5p se redujo, lo que llevó a la inhibición de la proliferación, migración e invasión celular. Por el contrario, mejoró la apoptosis. Al apuntar a EGR1, miR-183-5p contrarrestó parcialmente el impacto de EGR1 en la migración, invasión y fibrosis en KF. Los hallazgos implican que miR-183-5p suprime la formación de queloides al apuntar a EGR1. Como resultado, EGR1 es prometedor como objetivo terapéutico potencial para prevenir y tratar los queloides.

Pathogenic characteristics of bloodstream infections in patients with hematological diseases and the impact of stem cell transplantation on them / 中国热带医学

@#Abstract: Objective To investigate the epidemiological characteristics of pathogens causing bloodstream infection in hematology patients during treatment and to compare the effects of allogeneic hematopoietic stem cell transplantation (HSCT) on them, so as to provide evidence for the diagnosis and treatment of bloodstream infection. Methods A total of 292 cases with bloodstream infection in hematology wards of the PLA General Hospital were collected from 2017 to 2021, which were divided into HSCT group and N-HSCT group according to whether performed HSCT or not. The epidemiological characteristics and influence of pathogenic bacteria in blood stream infection were analyzed and compared between the two groups. Results A total of 362 strains of pathogenic bacteria were collected from 292 cases, including 106 strains in HSCT group (84 cases) and 256 strains in N-HSCT group (208 cases). Bloodstream infections were more common in acute myeloid leukemia (130/392, 44.52%), followed by non-Hodgkin's lymphoma (74/292, 25.34%). The rate of once bloodstream infection in HSCT group was higher than that in N-HSCT Group, but the rate of twice bloodstream infections in N-HSCT group was higher. Gram-negative Bacilli were the most common pathogens (56.08%), with Escherichia coli being absolutely dominant (109/362, 30.11%), followed by Klebsiella pneumoniae (39/362, 10.77%). Coagulase-negative staphylococci (CoNS) (107/362, 29.56%) were the most common Gram-positive cocci. The detection rate of fungi in HSCT group (10/106, 9.43%) was significantly higher than that in N-HSCT Group (3.52%). The drug resistance rate of the common pathogenic bacteria was at a high level, and there was a certain proportion of multi-drug resistant strains (except for Pseudomonas aeruginosa). The resistance rates of CoNS to penicillin, gentamicin, moxifloxacin, clindamycin and rifampicin in HSCT group were higher than those in N-HSCT Group. The resistance rate of Escherichia coli to piperacillin/tazobactam, cephalosporins and etapenem in HSCT group was significantly higher than that in N-HSCT group. Conclusions The pathogens of blood stream infection in hematology patients are complicated and various. It is difficult for clinical diagnosis and treatment to detect multiple infections and multiple pathogens. HSCT patients have a higher risk of fungal bloodstream infection and more multi-drug resistant strains detected. Therefore, the identification of bloodstream infection and multi-drug resistant strains associated with HSCT patients should prompt surveillance.

Comparison of primary keloid fibroblast cultivation methods and the characteristics of fibroblasts cultured from keloids, keloid-surrounding tissues, and normal skin tissues / Comparación de los métodos de cultivo de fibroblastos queloides primarios y las características de los fibroblastos cultivados a partir de queloides, tejidos circundantes queloides y tejidos cutáneos normales

SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes α -SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts.

RESUMEN: La identificación de un cultivo de fibroblastos queloides primarios, siempre ha sido una medida fundamental para estudiar los mecanismos de la enfermedad queloide. La calidad del cultivo de células primarias puede afectar directamente los resultados de otros experimentos. Este estudio se realizó para investigar las condiciones óptimas de crecimiento, incluido el tiempo óptimo de almacenamiento y el tiempo de tratamiento con colagenasa, para modelos de cultivo celular in vitro y los métodos adecuados para la separación epidermis-dermis en diferentes tejidos. Se recogieron de los pacientes tejidos queloides, tejidos circundantes queloides y tejidos cutáneos normales, para cultivo primario de fibroblastos. Se implementaron y compararon dos métodos, explante de tejido y digestión con colagenasa. Los niveles de expresión de los genes relacionados con queloides α -SMA, Col1 y Col3 se evaluaron en células cultivadas usando ambos métodos, para verificar las cualidades de las células primarias. Se realizó un análisis comparativo entre los dos métodos y entre los tres tejidos diferentes utilizados. La contaminación de bacterias y lípidos se minimizó inmediatamente después de que se procesaron las muestras. Se requirieron varios métodos de eliminación de la epidermis y diferentes tiempos de digestión con colagenasa en los tejidos para generar resultados óptimos. Los resultados de la PCR en tiempo real mostraron que los niveles de expresión de ARNm de genes relacionados con queloides en fibroblastos cultivados se correlacionaban con su perfil de expresión in vivo, como se informó en estudios anteriores. Los resultados de este studio indicaron varios puntos clave en el cultivo de fibroblastos queloides primarios y han demostrado la correlación en la expresión génica entre fibroblastos queloides in vivo y fibroblastos queloides primarios in vitro.

Efficacy of modified endoscopic release in treatment of carpal tunnel syndrome / 中国内镜杂志

Objective To explore a new technology system with single-portal and without special equipment to complete the arthroscopic treatment of carpal tunnel syndrome. Methods All the patients with CTS were randomly divided into two groups, modified endoscopic carpal tunnel release (MECTR) group (22 cases) and open carpal tunnel release (OCTR) group (24 cases). Nine parameters were evaluated, which including operation time, intraoperative complications, two-point discrimination 3 months after the operation, hospitalization time, scar pain score, incision infection rate, each patient's symptom amelioration (Kelly grading), the time needed to resume normal lifestyle and activity, symptoms of sympathetic dystrophy. Results No significant difference was observed between the MECTR group and OCTR group in regard to incision infection rate, intraoperative complications, two-point discrimination,symptoms of sympathetic dystrophy and clinical symptoms amelioration. In comparison to OCTS, MECTR significantly decreased operation time, hospitalization time, scar pain score and the time needed to resume normal lifestyle and activity. Conclusion MECTR for treatment of carpal tunnel syndrome has higher patient satisfaction, shorter operation time and hospitalization time, earlier return to work or normal lifestyle, less postoperative scar pain, so it is an effective method for the treatment of idiopathic carpal tunnel syndrome.

Frontalis muscle flap suspension surgery for the treatment of blepharoptosis based on the anatomical study of the frontal muscle nerve in the third of the eyebrow / Cirugía de suspensión del colgajo muscular frontal en el tratamiento de blefaroptosis basado en el estudio anatómico del nervio del músculo occipitofrontal en el tercio de la ceja

The aim of this study was to investigate the course of the supraorbital nerve and temporal branch of the facial nerve, and to verify the clinical security of cutting the frontalis muscle flap to treat blepharoptosis in one-third of the eyebrow. Twenty cadavers were dissected. The relationship of the supraorbital nerve and the course of the frontotemporal branch of the facial nerve with the head and neck muscles was evaluated. Forty patients underwent clinical frontal muscular flap suspension surgery for the treatment of blepharoptosis. The postoperative curative and complication rates were determined. The courses of the supraorbital nerve and frontotemporal branch of the facial nerve were observed to determine a relatively safe area in one-third of the eyebrow. The average width of the zone was 25.0±3.5 mm. In forty cases, satisfactory results were achieved in correcting blepharoptosis by cutting the frontal muscular flap in the middle of eyebrow within the wide range of 17±2.1 mm. No secondary sensory and motor dysfunctions occurred. One-third of the eyebrow (eyebrow center, within 17±2.1 mm) was a relatively safe area and allowed for the prevention of damage to the temporal branch of the facial nerve inside the supraorbital nerve and supraorbital artery and the outer frontotemporal branch of the facial nerve.

El objetivo de este estudio fue investigar el curso del nervio supraorbital y la rama temporal del nervio facial, para verificar la seguridad clínica de cortar el vientre frontal del músculo occipitofrontal (colgajo de músculo frontal) para tratar la blefaroptosis en un tercio de la ceja. Veinte cadáveres fueron disecados. Se evaluó la relación del nervio supraorbital y el curso de la rama temporal del nervio facial con los músculos de la cabeza y cuello. Cuarenta pacientes fueron sometidos a la cirugía de confección del colgajo del músculo frontal para el tratamiento de la ptosis palpebral. Se determinaron las tasas de curación y de complicaciones postoperatorias. Se observaron los cursos del nervio supraorbital y la rama temporal del nervio facial para determinar un área relativamente segura en un tercio de la ceja. El ancho medio de la zona fue 25,0±3,5 mm. En cuarenta casos, se lograron resultados satisfactorios en la corrección de la blefaroptosis con el colgajo del músculo frontal en la mitad de la ceja en un rango de 17±2,1 mm. No se produjeron disfunciones sensoriales o motoras secundarias. El tercio de la ceja (centro del entrecejo, dentro de 17±2,1 mm) es una zona relativamente segura y permite la prevención de daños al ramo temporal del nervio facial ubicada medial al nervio supraorbitario y a la arteria supraorbitaria, además del ramo temporal lateral del nervio facial.

Anti-HCMV and KSHV effect of a trapping ligand antagonist for Herpesvirus-encoded GPCR.

We have found and synthesized a trapping ligand peptide H22-LP (the conservative sequence is NAHCALL) from a random phage library according to the broad-spectrum trapping receptor H22, which derived from the residue 14-35 near the N-terminal region of receptor US28 on HCMV. In this study, we will evaluate its potential as an efficient antagonist of US28 and the anti-virus activity, acting as a broad spectrum chemokine receptors antagonist. Stable expression of US28 and ORF74 in NIH/3T3 cells were successfully constructed in vitro. Flow cytomety was used to determine the concentration of Ca2+ induced by H22-LP, and the binding of H22-LP and US28 was confirmed by enzyme-linked immunosorbent assay (ELISA). Antivirus activity of H22-LP on HCMV and KSHV was evaluated by anti-virus experiments. Our data suggest that H22-LP is an effectual antagonist of receptor US28 of HCMV and ORF74 of KSHV in the transfection assay, and it has potential to inhibit infection of HCMV and KSHV. These results provide support for the development of anti-virus strategies based on targeted inhibiting the infection of herpesvirus.