RESUMEN
Objective: The investigate the effects and mechanism of exosomes derived from human umbilical vein endothelial cells (HUVECs) on wound healing in diabetes rabbits. Methods: The experimental research methods were used. The primary vascular endothelial cells (VECs) and human skin fibroblasts (HSFs) were extracted from skin tissue around ulcer by surgical excision of two patients with diabetic ulcer (the male aged 49 years and the female aged 58 years) admitted to Xiangya Third Hospital of Central South University in June 2019. The cells were successfully identified through morphological observation and flow cytometry. The HUVEC exosomes were extracted by ultracentrifugation and identified successfully by morphological observation, particle size detection, and Western blotting detection. Twenty female 3-month-old New Zealand rabbits were taken to create one type 2 diabetic full-thickness skin defect wound respectively on both sides of the back. The wounds were divided into exosomes group and phosphate buffer solution (PBS) group and treated accordingly, with 20 wounds in each group, the time of complete tissue coverage of wound was recorded. On PID 14, hematoxylin-eosin staining or Masson staining was performed to observe angiogenesis or collagen fiber hyperplasia (n=20). The VECs and HSFs were co-cultured with HUVEC exosomes for 24 h to observe the uptake of HUVEC exosomes by the two kinds of cells. The VECs and HSFs were divided to exosome group treated with HUVEC exosomes and PBS group treated with PBS to detect the cell proliferation on 4 d of culture with cell count kit 8, to detect and calculate the cell migration rate at 24 and 48 h after scratch by scratch test, to detect the cell migration number at 24 h of culture with Transwell test, and to detect the mRNA expressions of nuclear factor-erythroid 2-related factor 2 (NRF2) and transcription activating factor 3 (ATF3) by real time fluorescence quantitative reverse transcription polymerase chain reaction. Besides, the number of vascular branches and vascular length were observed in the tube forming experiment after 12 h of culture of VECs (n=3). The VECs and HSFs were taken and divided into PBS group and exosome group treated as before, and NRF2 interference group, ATF3 interference group, and no-load interference group with corresponding gene interference. The proliferation and migration of the two kinds of cells, and angiogenesis of VECs were detected as before (n=3). Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and least significant difference test. Results: The time of complete tissue coverage of wound in exosome group was (17.9±1.9) d, which was significantly shorter than (25.2±2.3) d in PBS group (t=4.54, P<0.05). On PID14, the vascular density of wound in PBS group was significantly lower than that in exosome group (t=10.12, P<0.01), and the collagen fiber hyperplasia was less than that in exosome group. After 24 h of culture, HUVEC exosomes were successfully absorbed by VECs and HSFs. The proliferative activity of HSFs and VECs in exosome group was significantly higher than that in PBS group after 4 d of culture (with t values of 54.73 and 7.05, respectively, P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs (with t values of 3.42 and 11.87, respectively, P<0.05 or P<0.01) and VECs (with t values of 21.42 and 5.49, respectively, P<0.05 or P<0.01) in exosome group were significantly higher than those in PBS group. After 24 h of culture, the migration numbers of VECs and HSFs in exosome group were significantly higher than those in PBS group (with t values of 12.31 and 16.78, respectively, P<0.01). After 12 h of culture, the mRNA expressions of NRF2 in HSFs and VECs in exosome group were significantly higher than those in PBS group (with t values of 7.52 and 5.78, respectively, P<0.05 or P<0.01), and the mRNA expressions of ATF3 were significantly lower than those in PBS group (with t values of 13.44 and 8.99, respectively, P<0.01). After 12 h of culture, the number of vascular branches of VECs in exosome group was significantly more than that in PBS group (t=17.60, P<0.01), and the vascular length was significantly longer than that in PBS group (t=77.30, P<0.01). After 4 d of culture, the proliferation activity of HSFs and VECs in NRF2 interference group was significantly lower than that in PBS group and exosome group (P<0.05 or P<0.01); the proliferation activity of HSFs and VECs in ATF3 interference group was significantly higher than that in PBS group (P<0.05 or P<0.01) and significantly lower than that in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in ATF3 interference group were significantly higher than those in PBS group (P<0.05 or P<0.01) and significantly lower than those in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in NRF2 interference group were significantly lower than those in PBS group and exosome group (P<0.05 or P<0.01). After 24 h of culture, the migration numbers of VECs and HSFs in ATF3 interference group were significantly more than those in PBS group (P<0.05) and significantly less than those in exosome group (P<0.05 or P<0.01); the migration numbers of VECs and HSFs in NRF2 interference group were significantly less than those in PBS group and exosome group (P<0.01). After 12 h of culture, the vascular length and number of branches of VECs in NRF2 interference group were significantly decreased compared with those in PBS group and exosome group (P<0.01); the vascular length and number of branches of VECs in ATF3 interference group were significantly increased compared with those in PBS group (P<0.01) and were significantly decreased compared with those in exosome group (P<0.01). Conclusions: HUVEC exosomes can promote the wound healing of diabetic rabbits by promoting the proliferation and migration of VECs and HSFs, and NRF2 and ATF3 are obviously affected by exosomes in this process, which are the possible targets of exosome action.
Asunto(s)
Diabetes Mellitus , Exosomas , Animales , Femenino , Humanos , Masculino , Conejos , Colágeno/metabolismo , Exosomas/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Hiperplasia/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , ARN Mensajero/metabolismo , Úlcera , Cicatrización de Heridas , Persona de Mediana EdadRESUMEN
Objective: To investigate the efficacy and safety of a new cervical artificial disc prosthesis in the treatment of cervical degenerative diseases. Methods: The clinical data of 18 patients with single-level cervical degenerative diseases who underwent three dimensional printed anatomical bionic cervical disc replacement at Department of Spinal Surgery,Honghui Hospital,Xi'an Jiaotong University from May 2019 to July 2020 were analyzed retrospectively. There were 7 males and 11 females,aged (45±8) years old(range:28 to 58 years).The surgical segment was located at C3ï¼4 level in 2 cases, C4ï¼5 level in 5 cases, C5ï¼6 level in 9 cases, and C6ï¼7 level in 2 cases.The clinical and radiographic outcomes were recorded and compared at preoperative,postoperative times of one month and twelve months.The clinical assessments contained Japanese orthopedic association (JOA) score,neck disability index (NDI) and visual analogue scale (VAS).Imaging assessments included range of motion (ROM) of cervical spine, prosthesis subsidence and prosthesis anteroposterior migration.Repeated measurement variance analysis was used for comparison between groups,and paired t test was used for pairwise comparison. Results: All patients underwent the operation successfully and were followed up for more than 12 months.Compared with preoperative score,the JOA score,NDI and VAS were significantly improved after surgery (all P<0.01).There was no significant difference in postoperative ROM compared with 1-and 12-month preoperative ROM (t=1.570,P=0.135;t=1.744,P=0.099). The prosthesis subsidence was (0.29±0.13) mm (range: 0.18 to 0.50 mm) at 12-month postoperatively.The migration of prosthesis at 12-months postoperatively were (0.71±0.20) mm (range: 0.44 to 1.08 mm).There was no prosthesis subsidence or migration>2 mm at 12-month postoperatively. Conclusion: Three dimensional printed anatomical biomimetic cervical artificial disc replacement has a good early clinical effect in the treatment of cervical degenerative diseases, good mobility can be obtained while maintaining stability.
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Degeneración del Disco Intervertebral , Disco Intervertebral , Reeemplazo Total de Disco , Adulto , Biomimética , Vértebras Cervicales/cirugía , Femenino , Estudios de Seguimiento , Humanos , Disco Intervertebral/cirugía , Degeneración del Disco Intervertebral/cirugía , Masculino , Persona de Mediana Edad , Rango del Movimiento Articular , Estudios Retrospectivos , Reeemplazo Total de Disco/métodos , Resultado del TratamientoRESUMEN
Whiteflies (Bemisia tabaci) are phloem feeders, and some invasive species are composed of cryptic species complexes that cause extensive crop damage, particularly via the direct transmission of plant viruses. Apoptosis is a type of programmed cell death essential for organismal development and tissue homeostasis. The caspases belong to a family of cysteine proteases that play a central role in the initiation of apoptosis in many organisms. Here, we employed a comprehensive genomics approach to identity caspases in B. tabaci Middle East Asia Minor 1 (MEAM1), an invasive whitefly that carries a cryptic species complex that is devastating to crops. Four caspase genes were identified, and their motif compositions were predicted. Structures were relatively conserved in both putative effector and initiator caspases. Expression patterns of caspase genes differed across insect developmental stages. Three caspase genes were induced immediately after ultraviolet (UV) treatment. Expression levels of Bt-caspase-1 and Bt-caspase-3b increased in the midgut and salivary glands during apoptosis induced by UV treatments, whereas silencing of both genes reduced UV-triggered apoptosis. Our study demonstrates that Bt-caspase-1 and Bt-caspase-3b, respectively, act as putative initiator and effector apoptotic caspases in the MEAM1 whitefly.
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Apoptosis , Caspasas/metabolismo , Hemípteros/efectos de la radiación , Secuencia de Aminoácidos , Animales , Caspasas/genética , Hemípteros/enzimología , Hemípteros/genética , Filogenia , Dominios Proteicos , Rayos UltravioletaRESUMEN
TNF alpha induced protein 3 (TNFAIP3), a member of zinc finger protein family, is a gene whose expression level is promptly induced by the tumor necrosis factor. In this study, the clinical significance of TNFAIP3 was analyzed based on available samples in The Cancer Genome Atlas database. TNFAIP3 downregulation was associated with distant metastasis and worse patient prognosis. TNFAIP3-overexpressing and TNFAIP3-knockdown NPC cell line models were established through plasmid-mediated overexpression and small interfering RNA (siRNA), respectively. Cell migration and invasion capacities were evaluated by wound-healing and transwell assays. Functional studies indicated that TNFAIP3 knockdown promoted migration and invasion, whereas TNFAIP3 overexpression alleviated these functions. Western blot analysis was used to examine protein changes from TNFAIP3 overexpression and knockdown, in which TNFAIP3 promoted the protein expression of E-cadherin and suppressed vimentin expression. Our data suggested that TNFAIP3 inhibited migration and invasion by suppressing epithelial mesenchymal transition in NPC.
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Transición Epitelial-Mesenquimal , Carcinoma Nasofaríngeo/genética , Neoplasias Nasofaríngeas/genética , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa/genética , Antígenos CD/metabolismo , Cadherinas/metabolismo , Línea Celular Tumoral , Movimiento Celular , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Invasividad Neoplásica , ARN Interferente Pequeño , Vimentina/metabolismoRESUMEN
OBJECTIVE: Sepsis presents great threat to human health. Here we aimed to understand its pathogenesis and discover effective therapeutic targets. MATERIALS AND METHODS: Gene expression data for sepsis samples was compared with healthy controls to identify differentially expressed genes (DEGs), then we constructed PPI network to detect the network clustering. Gene Ontology (GO) analysis was also done to identify over-represented biological pathways. RESULTS: KEGG pathway analysis revealed that PI3K-AKT signaling pathway, chemokine signaling pathway and MAPK signaling pathway were significantly over-represented in these DEGs. Given proteins work together to exert certain biological functions, network analysis was done to identify genes closely associated with these DEGs. Using Fisher's exact test, 8 genes such as NEDD8, CUL1 and CUL3 were screened out. CONCLUSIONS: Overall, our findings not only supplement the knowledge about sepsis, but also provide a number of potential biomarkers for diagnosis and treatment.
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Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Sepsis/genética , Estudios de Casos y Controles , Perfilación de la Expresión Génica , Humanos , Sistema de Señalización de MAP Quinasas , Fosfatidilinositol 3-Quinasas/metabolismo , Sepsis/metabolismo , Sepsis/patología , Transducción de SeñalRESUMEN
A total of 28 patients with clinically localized prostate cancer (PCa) underwent extraperitoneal laparoscopic radical prostatectomy (EP-LRP). The mean operative duration was 309 (287-600) minutes. Estimated blood loss ranged from 380 to 1000 (mean 480) ml. At 3 to 5 days postoperatively, the catheter was removed. No open conversion was required and no patient presented postoperative complications. PSA level was less than 0.1 ng/ml at 3 months after surgery in all patients. At a mean follow-up of 10 (6-16) months, there were no biochemical failures. The extraperitoneal technique potentially decreased the risk of intra-abdominal complications and better approximated than open retropubic radical prostatectomy. In conclusion, EP-LRP is an effective, safe and precise technique.
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Laparoscopía/métodos , Prostatectomía/métodos , Neoplasias de la Próstata/cirugía , Humanos , Masculino , Complicaciones PosoperatoriasRESUMEN
AIM: To put forward criteria for the pressure assessment in the operation of intercavernous embedding of bulboperineal urethra for the treatment of urinary incontinence after prostatic operation. METHODS: A F14 urethral catheter is inserted during the operation and upon suturing the corpora cavemosa centrally, the catheter is slowly pushed in and pulled out in order that the operator feels a certain degree of close-fit resistance. The degree of tightness of the stitches, which regulate the compression pressure, is adjusted in accordance with this close-fit sensation. To further ascertain the adequacy of the force of compression, the bladder is filled with 300 ml physiological saline and observe the appropriateness (size and continuity) of the outflow stream when the lower abdomen is depressed with a pressure of 80-90 cm H2O. The operation was given to six patients suffered from urinary incontinence for 20 or more months after prostatic operation. RESULTS: Five cases achieved complete recovery, while the therapeutic effect of the 6th one was not satisfactory. A second stage operation was carried out 3 months later with the addition of one more stitch both proximally and distally to reinforce the compression force. The condition was improved dramatically. The follow-up period averaged 3.5 years. CONCLUSION: The adequacy of the compression pressure exerted by the juxtaposed corpora cavernosa is the key point determining the outcome of the operation. The measures for assessing the compression pressure suggested by the authors are helpful in obtaining the good results of the present paper (6/6 success) as compared with 25/34 success in the previous report.
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Complicaciones Posoperatorias/cirugía , Hiperplasia Prostática/cirugía , Uretra/cirugía , Incontinencia Urinaria/cirugía , Anciano , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Presión , Cateterismo Urinario/métodos , Urodinámica , Procedimientos Quirúrgicos Urológicos Masculinos/métodosRESUMEN
We studied 128 patients with active peptic ulcer diagnosed by gastro-endoscopy four weeks after treatment with Xi Lei powder, PGE2 levels in both serum and gastro-duodenal mucosa were significantly higher than that before the treatment, the difference was significant (P < 0.01). The rate of negative conversion of Helicobacter pylori (HP) showed in 63.3% of cases. The distributed density of HP significantly reduced, the difference was significant (P < 0.01). These results indicated that Xi Lei powder could heal ulcer probably owing to the increasing of the PGE2 level in gastroduodenal mucosa, it inhibited the growth and reproduction of HP, and eliminated the HP, but did not inhibit the secretion of gastric acid and release the gastrin.