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This study aims to explore the molecular mechanisms of miR-206-3p in regulating Hsp90aa1 and its involvement in the central nervous system (CNS) injury in heat stroke. Weighted gene co-expression network analysis (WGCNA) was performed on the GSE64778 dataset of heat stroke to identify module genes most closely associated with disease characteristics. Through the selection of key genes and predicting upstream miRNAs using RNAInter and miRWalk databases, the regulatory relationship between miR-206-3p and Hsp90aa1 was determined. Through in vitro experiments, various methods, including bioinformatics analysis, dual-luciferase reporter gene assay, RIP experiment, and RNA pull-down experiment, were utilized to validate this regulatory relationship. Furthermore, functional experiments, including CCK-8 assay to test neuron cell viability and flow cytometry to assess neuron apoptosis levels, confirmed the role of miR-206-3p. Transmission electron microscopy, real-time quantitative PCR, DCFH-DA staining, and ATP assay were employed to verify neuronal mitochondrial damage. Heat stroke rat models were constructed, and mNSS scoring and cresyl violet staining were utilized to assess neural functional impairment. Biochemical experiments were conducted to evaluate inflammation, brain water content, and histopathological changes in brain tissue using H&E staining. TUNEL staining was applied to detect neuronal apoptosis in brain tissue. RT-qPCR and Western blot were performed to measure gene and protein expression levels, further validating the regulatory relationship in vivo. Bioinformatics analysis indicated that miR-206-3p regulation of Hsp90aa1 may be involved in CNS injury in heat stroke. In vivo, animal experiments demonstrated that miR-206-3p and Hsp90aa1 co-localized in neurons of the rat hippocampal CA3 region, and with prolonged heat stress, the expression of miR-206-3p gradually increased while the expression of Hsp90aa1 gradually decreased. Further in vitro cellular mechanism validation and functional experiments confirmed that miR-206-3p could inhibit neuronal cell viability and promote apoptosis and mitochondrial damage by targeting Hsp90aa1. In vivo, experiments confirmed that miR-206-3p promotes CNS injury in heat stroke. This study revealed the regulatory relationship between miR-206-3p and Hsp90aa1, suggesting that miR-206-3p could regulate the expression of Hsp90aa1, inhibit neuronal cell viability, and promote apoptosis, thereby contributing to CNS injury in heat stroke.
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BACKGROUND: Deep learning provides an efficient automatic image recognition method for small bowel (SB) capsule endoscopy (CE) that can assist physicians in diagnosis. However, the existing deep learning models present some unresolved challenges. AIM: To propose a novel and effective classification and detection model to automatically identify various SB lesions and their bleeding risks, and label the lesions accurately so as to enhance the diagnostic efficiency of physicians and the ability to identify high-risk bleeding groups. METHODS: The proposed model represents a two-stage method that combined image classification with object detection. First, we utilized the improved ResNet-50 classification model to classify endoscopic images into SB lesion images, normal SB mucosa images, and invalid images. Then, the improved YOLO-V5 detection model was utilized to detect the type of lesion and its risk of bleeding, and the location of the lesion was marked. We constructed training and testing sets and compared model-assisted reading with physician reading. RESULTS: The accuracy of the model constructed in this study reached 98.96%, which was higher than the accuracy of other systems using only a single module. The sensitivity, specificity, and accuracy of the model-assisted reading detection of all images were 99.17%, 99.92%, and 99.86%, which were significantly higher than those of the endoscopists' diagnoses. The image processing time of the model was 48 ms/image, and the image processing time of the physicians was 0.40 ± 0.24 s/image (P < 0.001). CONCLUSION: The deep learning model of image classification combined with object detection exhibits a satisfactory diagnostic effect on a variety of SB lesions and their bleeding risks in CE images, which enhances the diagnostic efficiency of physicians and improves the ability of physicians to identify high-risk bleeding groups.
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Aprendizado Profundo , Humanos , Intestino Delgado/diagnóstico por imagem , Intestino Delgado/patologiaRESUMO
PURPOSE: Accurate prediction of extraprostatic extension (EPE) is pivotal for surgical planning. Herein, we aimed to provide an updated model for predicting EPE among patients diagnosed with MRI-targeted biopsy. MATERIALS AND METHODS: We analyzed a multi-institutional dataset of men with clinically localized prostate cancer diagnosed by MRI-targeted biopsy and subsequently underwent prostatectomy. To develop a side-specific predictive model, we considered the prostatic lobes separately. A multivariable logistic regression analysis was fitted to predict side-specific EPE. The decision curve analysis was used to evaluate the net clinical benefit. Finally, a regression tree was employed to identify three risk categories to assist urologists in selecting candidates for nerve-sparing, incremental nerve sparing and non-nerve-sparing surgery. RESULTS: Overall, data from 3169 hemi-prostates were considered, after the exclusion of prostatic lobes with no biopsy-documented tumor. EPE was present on final pathology in 1,094 (34%) cases. Among these, MRI was able to predict EPE correctly in 568 (52%) cases. A model including PSA, maximum diameter of the index lesion, presence of EPE on MRI, highest ISUP grade in the ipsilateral hemi-prostate, and percentage of positive cores in the ipsilateral hemi-prostate achieved an AUC of 81% after internal validation. Overall, 566, 577, and 2,026 observations fell in the low-, intermediate- and high-risk groups for EPE, as identified by the regression tree. The EPE rate across the groups was: 5.1%, 14.9%, and 48% for the low-, intermediate- and high-risk group, respectively. CONCLUSION: In this study we present an update of the first side-specific MRI-based nomogram for the prediction of extraprostatic extension together with updated risk categories to help clinicians in deciding on the best approach to nerve-preservation.
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Biópsia Guiada por Imagem , Imageamento por Ressonância Magnética , Prostatectomia , Neoplasias da Próstata , Humanos , Masculino , Neoplasias da Próstata/patologia , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias da Próstata/cirurgia , Imageamento por Ressonância Magnética/métodos , Biópsia Guiada por Imagem/métodos , Pessoa de Meia-Idade , Idoso , Prostatectomia/métodos , Próstata/patologia , Próstata/diagnóstico por imagem , Próstata/cirurgia , Nomogramas , Prognóstico , Estudos Retrospectivos , Gradação de TumoresRESUMO
Objective:To study the expression of microRNA (miRNA)-4783-3p in liver cancer tissue and its effect on the proliferation and migration of liver cancer Huh-7 cells.Methods:The cBioPortal database was used to analyze the expression of miR-4783-3p in liver cancer tissues and adjacent tissues. In strict accordance with the instructions of Lipofectamine? 2000 transfection kit, miR-4783-3p overexpression mimics or overexpression control mimics were transfected into Huh-7 cells respectively, namely overexpression group and control group. The proliferation of Huh-7 cells was analyzed by CCK-8 assay, and the migration of Huh-7 cells was analyzed by cell scratch method. The targeting relationship between miR-4783-3p and insulin-like growth factor binding protein 2 ( IGFBP2) mRNA was detected by dual-luciferase reporter gene assay. RT-qPCR was used to detect the expression of IGFBP2 mRNA. Western-blotting was used to detect the expression of IGFBP2 protein and EGFR-STAT3 molecular pathway proteins. Results:The expression of miR-4783-3p in liver cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). Compared with the control group, the proliferation ability of Huh-7 cells in the overexpression group was significantly decreased ( P<0.05). The scratch healing rates of Huh-7 cells in the control group and the overexpression group were (67.71±9.04)% and (29.58±10.51)%, respectively, and the scratch healing rate in the overexpression group was significantly lower ( P<0.01). miR-4783-3p targeted and bound to IGFBP2 mRNA ( P<0.01). The expression of IGFBP2 mRNA in the control and overexpression groups was 5.76±1.44 and 0.99±0.47, respectively, and miR-4783-3p negatively regulated the expression of IGFBP2 mRNA ( P<0.01). Compared with the control group, the expressions of IGFBP2 protein and EGFR-STAT3 molecular pathway protein were decreased in the overexpression group. Conclusions:miR-4783-3p is lowly expressed in liver cancer tissues. miR-4783-3p can attenuate the proliferation and invasion ability of liver cancer Huh-7 cells by inducing the low expression of IGFBP2 gene.
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OBJECTIVE@#To summarize the biomechanical characteristics, diagnosis, and hip arthroscopic treatment of borderline developmental dysplasia of hip (BDDH) with Cam-type femoroacetabular impingement (Cam FAI).@*METHODS@#The literature on BDDH with Cam FAI at home and abroad in recent years was extensively reviewed and analyzed.@*RESULTS@#In patients with BDDH and Cam FAI, the femoral neck anteversion angle and femoral neck shaft angle increase, the pelvis tilts, and the acetabulum rotates, resulting in instability of the hip joint. In order to maintain the stability of the hip joint, the direction of biomechanical action of the hip joint has changed, which further affects the anatomical structures such as the proximal femur and acetabular morphology. BDDH with Cam FAI can be diagnosed clinically by combining lateral center edge angle, anterior center edge angle, and acetabular index. BDDH with Cam FAI can be effectively treated through arthroscopic polishing of the edges of the acetabular proliferative bone, excision of Cam malformations, and minimally invasive repair of the glenoid lip and cartilage of the hip joint.@*CONCLUSION@#Currently, there is no unified standard for the diagnosis and treatment of BDDH with Cam FAI. Minimally invasive treatment of the hip under arthroscopy can achieve good early- and medium-term effectiveness, and has certain advantages in repairing and maintaining the integrity of the glenoid lip and suturing/compression joint capsule. However, the long-term effectiveness needs to be further followed up to determine. The timing of surgery, intraoperative bone edge depth polishing, and joint capsule suturing/compression techniques also need to be further explored.
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Humanos , Impacto Femoroacetabular/cirurgia , Artroscopia/métodos , Articulação do Quadril/cirurgia , Acetábulo/cirurgia , Luxação Congênita de Quadril/cirurgia , Resultado do Tratamento , Estudos RetrospectivosRESUMO
Objective: To explore the impact of health management measures for entry personnel (entry management measures) against COVID-19 on the epidemiological characteristics of imported Dengue fever in Guangdong Province from 2020 to 2022. Methods: Data of imported Dengue fever from January 1, 2016 to August 31, 2022, mosquito density surveillance from 2016 to 2021, and international airline passengers and Dengue fever annual reported cases from 2011 to 2021 in Guangdong were collected. Comparative analysis was conducted to explore changes in the epidemic characteristics of imported Dengue fever before the implementation of entry management measures (from January 1, 2016 to March 20, 2020) and after the implementation (from March 21, 2020 to August 31, 2022). Results: From March 21, 2020, to August 31, 2022, a total of 52 cases of imported Dengue fever cases were reported, with an imported risk intensity of 0.12, which were lower than those before implementation of entry management measures (1 828, 5.29). No significant differences were found in the characteristics of imported cases before and after implementation of entry management measures, including seasonality, sex, age, career, and imported countries (all P>0.05). 59.62% (31/52) of cases were found at the centralized isolation sites and 38.46% (20/52) at the entry ports. However, before implementation of entry management measures, 95.08% (1 738/1 828) of cases were found in hospitals. Among 51 cases who had provided entry dates, 82.35% (42/51) and 98.04% (50/51) of cases were found within seven days and fourteen days after entry, slightly higher than before implementation [(72.69%(362/498) and 97.59% (486/498)]. There was significant difference between the monthly mean values of Aedes mosquito larval density (Bretto index) from 2020 to 2021 and those from 2016 to 2019 (Z=2.83, P=0.005). There is a strong positive correlation between the annual international airline passengers volume in Guangdong from 2011 to 2021 and the annual imported Dengue fever cases (r=0.94, P<0.001), and a positive correlation also existed between the international passenger volume and the annual indigenous Dengue fever cases (r=0.72, P=0.013). Conclusions: In Guangdong, the entry management measures of centralized isolation for fourteen days after entry from abroad had been implemented, and most imported Dengue fever cases were found within fourteen days after entry. The risk of local transmission caused by imported cases has reduced significantly.
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Animais , Humanos , COVID-19 , Aedes , Epidemias , China/epidemiologia , Dengue/epidemiologiaRESUMO
@#Abstract: Objective - To understand the status of occupational hazards in non coal mine mountains in Hunan Province. Methods - - A cross sectional survey was conducted on 432 non coal mining enterprises in Hunan Province in 2021. The field , occupational health survey occupational disease hazard factors detection and occupational health monitoring data analysis were Results - carried out. Limited liability companies and private enterprises were the main economic types of non coal mining , , mountains in Hunan Province accounting for 55.3% and 32.4% respectively. The size of enterprises was mainly small and , ; - micro enterprises accounting for 59.3% and 35.2% respectively 78.0% of the enterprises were in non metallic mining. The - , over standard rates of silica dust and noise in workplaces were 17.1% and 29.4% respectively. The exposure rate of , (P ) occupational hazard factor was 47.3%. The smaller the enterprise scale the higher the exposure rate <0.01 . The number of , , people exposed to silica dust and noise was the largest with the exposure rate of 29.4% and 31.0% respectively. The rates of , occupational health training for persons in charge of enterprises occupational health management personnel and workers were , , , 73.8% 73.4% and 85.0% respectively. The smaller the enterprise scale the lower the occupational health training rates of , , enterprise leaders occupational health management personnel and workers and the lower the implementation rates of ( P ) - - enterprise occupational health examination all <0.05 . The fully installed rate and effective rate of occupational disease ; prevention facilities were 6.9% and 1.6% respectively and the fully deployed rate and effective wearing rate of personal , protective equipment were 11.1% and 6.2% respectively. The detection rates of occupational contraindications and suspected , Conclusion occupational diseases in workers exposed to silica dust and noise were 2.0% and 2.9% respectively. The - , occupational risks such as silica dust and noise are serious in non coal mine mountains of Hunan Province and the foundation of occupational health management is weak in small and micro enterprises.
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Objective:To analyze the expression and clinical significance of reticulocalbin 3 (RCN3) in colon cancer by bioinformatics database and biological experiments.Methods:Colon cancer HT29 and SW620 cells and colon normal mucosal cells FHC were cultured. The expression of RCN3 in cells was verified by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. The expression data of RCN3 in normal colon tissue and colon cancer tissue were obtained by Ualcan database. The co-expressed gene information of RCN3 from LinkedOmics database was obtained, and the biological processes and related functions of these RCN3 co-expressed genes through were analyzed by gene ontology analysis (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The protein-protein interaction network of RCN3 related coding genes was constructed by using STRING database. Finally, the relationship between the expression of RCN3 and the clinical prognosis of patients with colon cancer was compared and analyzed according to GEPIA, Ualcan and Linked Omics biological database.Results:Western blot and qRT-PCR results showed that the mRNA and protein expression of RCN3 in HT29 and SW620 colon cancer cells was significantly higher than those in FHCcells ( all P<0.05). The analysis of biological database showed that the expression level of RCN3 in colon cancer tissue was higher than that in normal colon tissue ( P<0.05). GO enrichment analysis showed that RCN3 co-expression genes were mainly involved in the composition of extracellular matrix and extracellular domain structure, the binding process of extracellular matrix and multiple receptors, and the biological processes related to tumor development such as cell adhesion, immune response, and angiogenesis through extracellular domain structure. KEGG pathway analysis showed that RCN3 co-expression genes were mainly involved in ECM receptor interaction, cytokine receptor interaction, chemokine signaling pathway, phosphatidylinositol-3-kinase protein kinase B (PI3K-Akt) signaling pathway, phagosome signal, IgA related intestinal immune network signal, these signaling pathways always related to tumor invasion, migration and inflammatory immune response. The protein-protein interaction network analysis showed that the coding protein genes that directly interacted with RCN3 protein that included PRDX6, NOSIP, PCSK6, IMMP1L, PRRG2, FBXO47, FCGRT, FKBP9, PCDHGA12, and PNMAL1, which were mainly involved in the occurrence and development of colorectal cancer, liver cancer, gastric cancer, breast cancer, lung cancer, and ovarian cancer. Survival curve analysis showed that the overall survival rate of colon cancer patients with high expression of RCN3 was significantly lower than that of patients with low expression of RCN3 ( P<0.05). Conclusions:RCN3 is highly expressed in colon cancer tissues and cells, which is closely related to the occurrence, development and prognosis of colon cancer. It can be used as one of the markers for early screening and prognosis prediction of colon cancer.
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Insulin therapy plays an essential role in the treatment of diabetes mellitus. However, frequent injections required to effectively control the glycemic levels lead to substantial inconvenience and low patient compliance. In order to improve insulin delivery, many efforts have been made, such as developing the nanoparticles (NPs)-based release systems and oral insulin. Although some improvements have been achieved, the ultimate results are still unsatisfying and none of insulin-loaded NPs systems have been approved for clinical use so far. Recently, nano‒protein interactions and protein corona formation have drawn much attention due to their negative influence on the
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Objective To study the expression of lipoic acid synthase(LIAS)in the liver and kidney of Leprdb/db mice with deficient leptin receptor. Methods Eight 10-week old male Leprdb/ +mice and Leprdb/dbmice were included in this study. The body weight of rats in the two groups was measured. Fasting blood glucose(FPG)was measured with blood glucose test strips for all mice after fasting for 8 hours. Blood samples were obtained from the abdominal aorta and the animals were sacrificed. The liver and kidney were weighed. The right lobe of liver and the left kidney samples were fixed in 4% paraformaldehyde for pathological examination. Serum samples were separated and the sereum contents of CHO, TG,HDL and LDL were detected. The mitochondria of liver and kidney tissues were extracted with a mitochondrial isolation kit, and the protein was extracted. The expression of LIAS protein was detected by western blot. Results Histopathological observation showed that the liver and kidney tissues of Leprdb/ +mice have intact and clear structure. But the liver tissue of Leprdb/dbmice showed fatty degeneration, the kidney tissue showed glomerular hypertrophy, basement membrane thickening, mesangial area widened, including mesangial cells and mesangial matrix increased. The GLU,CHO,TG,LDL and AST of Leprdb/dbmice were significantly increased compared with those of Leprdb/ +mice(P<0.05). Compared with Leprdb/ +mice,the LIAS protein expression was significantly increased in the liver and kidney mitochondria of Leprdb/dbmice(P<0.05). Conclusions There is impaired glucose and lipid metabolism in the Leprdb/dbmice which has defect leptin receptor,and the expression of LIAS protein in liver and kidney of the Leprdb/dbmice is higher than that of Leprdb/ +mice.
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Objective To establish an efficient method of genotyping for Leprdb/ +mouse offsprings by TaqMan probe quantitative fluorescence PCR. Methods Genome DNA was extracted from tails of 228 Leprdb/ +mouse offsprings. PCR primers and TaqMan probes were designed according to the mutation sites of Lepr gene(rs1801133). Real time PCR assay was applied and SNP loci were typed with SDS software. The genotyping of 2-month old Leprdb/dbmice was validated by the phenotype and Hardy-Weinberg equilibrium test was performed. Results 228 samples were detected by the established TaqMan fluorescence quantitative PCR assay. 64 mice were of GG genotype, with a genotype frequency of 0.1929. 123 mice were of GT genotype, with a genotype frequency of 0.5395. 41 mice were of TT genotype, with a genotype frequency of 0.2807. Compared with the phenotype typing,the sensitivity of the TaqMan fluorescence quantitative PCR was 97.56% and the specificity was 99.47%. Conclusions TaqMan probe quantitative fluorescence PCR assay is a simple and efficient method,and can be used to detect the genotype of Leprdb/ +mouse offsprings.
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<p><b>Objective</b>To explore the application value of real-time contrast-enhanced ultrasound (RTCEU) in improving the detection rate of transrectal ultrasound-guided prostate biopsy.</p><p><b>METHODS</b>This prospective study included 91 male patients with abnormally high PSA (4-20 μg/L) or abnormalities in DRE or MRI, who underwent 12+X prostate biopsy following conventional transrectal ultrasonography (TRUS) and RTCEU examination. We compared the numbers of suspected prostatic nodules before and after RTCEU as well as the detection rates of prostate cancer between conventional TRUS-guided 12PBx and 12PBx plus lesion-targeted biopsy procedures.</p><p><b>RESULTS</b>Totally, 57 of the 86 suspected lesions on TRUS (66.3%), and 108 of the 118 abnormal nodules on RTCEU (91.5%) were confirmed to be prostate cancer. RTCEU achieved a significantly higher detection rate than TRUS (P<0.01). A total of 39 cases of prostate cancer (42.8%) were detected by RTCEU, while only 28 (30.7%) by TRUS, with statistically significant difference in the detection rate between the two procedures (P=0.033).</p><p><b>CONCLUSIONS</b>Real-time contrast-enhanced ultrasound can significantly improve the detection rate of prostate cancer and provide a valuable guide to targeted prostate biopsy.</p>
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Humanos , Masculino , Meios de Contraste , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico , Imageamento por Ressonância Magnética , Estudos Prospectivos , Próstata , Diagnóstico por Imagem , Patologia , Antígeno Prostático Específico , Sangue , Neoplasias da Próstata , Sangue , Diagnóstico por Imagem , Patologia , Ultrassonografia de IntervençãoRESUMO
Osteoarthritis is recognised to be an interactive pathological process involving the cartilage, subchondral bone and synovium. The signals from the synovium play an important role in cartilage metabolism, but little is known regarding the influence of the signalling from bone. Additionally, the collagenases and stromelysin-1 are involved in cartilage catabolism through mitogen-activated protein kinase (MAPK) signalling, but the role of the gelatinases has not been elucidated. Here, we studied the influence of osteoclastic signals on chondrocytes by characterising the expression of interleukin-1β (IL-1β)-induced gelatinases through MAPK signalling. We found that osteoclast-conditioned media attenuated the gelatinase activity in chondrocytes. However, IL-1β induced increased levels of gelatinase activity in the conditioned media group relative to the mono-cultured chondrocyte group. More specifically, IL-1β restored high levels of gelatinase activity in c-Jun N-terminal kinase inhibitor-pretreated chondrocytes in the conditioned media group and led to lower levels of gelatinase activity in extracellular signal-regulated kinase or p38 inhibitor-pretreated chondrocytes. Gene expression generally correlated with protein expression. Taken together, these results show for the first time that signals from osteoclasts can influence gelatinase activity in chondrocytes. Furthermore, these data show that IL-1β restores gelatinase activity through MAPK inhibitors; this information can help to increase the understanding of the gelatinase modulation in articular cartilage.
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Animais , Camundongos , Células 3T3 , Cartilagem Articular , Biologia Celular , Sobrevivência Celular , Fisiologia , Células Cultivadas , Condrócitos , Técnicas de Cocultura , Meios de Cultivo Condicionados , Gelatinases , Interleucina-1beta , Farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Sistema de Sinalização das MAP Quinases , Fisiologia , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz , Proteínas Quinases Ativadas por Mitógeno , Monócitos , Biologia Celular , NF-kappa B , Osteoclastos , Fisiologia , Inibidores de Proteases , Inibidor Tecidual de Metaloproteinase-1 , Inibidor Tecidual de Metaloproteinase-2 , Proteínas Quinases p38 Ativadas por MitógenoRESUMO
To assess the postoperative outcomes of piggyback implantation using Acrysof Toric intraocular lens ( lOL ) in high myopia combined with corneal astigmatism. ●METHODS: Sixty patients who had phacoemulsification with lOL implantation due to high myopia, cataract and corneal astigmatism from January 2010 to June 2013 were randomly divided into observation group (piggyback Toric lOL implantation, both an Acrysoft lQ Toric lOL and a minus foldable acrylic three piece lOL were implanted in the capsular bag, n= 30) and control group (foldable lOL implantation, n = 30). Postoperative follow - up went on 6mo. lnformation collected included uncorrected visual acuity ( UCVA), lOL position, residual astigmatism and complications. ● RESULTS: The UCVA increased from 3. 52 ± 0. 03 preoperatively to 4. 78±0. 01 at 6mo postoperatively in the observation group, from 3. 51±0. 03 preoperatively to 4. 30± 0. 13 at 6mo postoperatively in the control group. The observation group's postoperative UCVA was better than that of the control group. There was statistically significant difference ( t = 3. 612, P ●CONCLUSlON: Piggyback implantation using Toric lOL can help to solve the problem of no matching Toric lOL power for the high myopia combined with corneal astigmatism at the current stage. lt improves the UCVA and reduces the astigmatism after the cataract surgery.
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<p><b>OBJECTIVE</b>To screen the lentiviral vector carrying siRNA and capable of significantly suppressing the ROCK2 gene expression in the corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).</p><p><b>METHODS</b>We designed and synthesized 4 siRNA fragments targeting the ROCK2 gene and packaged them into lentiviral vectors. We collected corpus cavernosum smooth muscle cell samples from 5 male SHRs and randomly divided them into groups A (non-transfection control), B (GFP lentiviral transfection), C, D, E, and F (lentiviral transfection with siRNA fragments 1 -4 targeting the ROCK2 gene). Each group consisted of 5 samples and each sample 3 x 10(4) cells. At 48 hours after transfecting MOI = 80 into the SHR corpus cavernosum smooth muscle cells, we detected the expression of GFP under the fluorescent microscope and the mRNA expression of the ROCK2 gene by RT-PCR.</p><p><b>RESULTS</b>The transfection efficiency of the SHR corpus cavernosum smooth muscle cells was > 50%. Compared with group A, the expression of ROCK2 mRNA in the corpus cavernosum smooth muscle cells showed no remarkable change in group B (P > 0.05) but was inhibited very significantly in C ([43.91 +/- 8.19]%), D ([47.15 +/- 6.64]%), and F ([25.7 +/- 6.03]%) (P < 0.01), and significantly in E ([16.81 +/- 5.94]%, P < 0.05).</p><p><b>CONCLUSION</b>We successfully constructed 4 lentiviral vectors carrying siRNA targeting the ROCK2 gene, all of which can significantly suppress the ROCK2 expression in the SHR corpus cavernosum smooth muscle cells, and one has a highly strong inhibitory effect.</p>
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Animais , Masculino , Ratos , Inativação Gênica , Vetores Genéticos , Lentivirus , Genética , Miócitos de Músculo Liso , Pênis , Biologia Celular , RNA Mensageiro , Metabolismo , RNA Interferente Pequeno , Distribuição Aleatória , Ratos Endogâmicos SHR , Transfecção , Métodos , Quinases Associadas a rho , GenéticaRESUMO
Objective To investigate the interventional treatment strategy for occluding the intercristal ventricular septal defect (VSD) in order to improve the surgical safety and success rate. Methods During the period from January 2012 to December 2013, a total of 31 patients with intercristal VSD were admitted to authors’ hospital to receive interventional catheter occlusion therapy. Preoperative color Doppler ultrasound echocardiography showed that on the short axis view of the aorta the VSD interrupted port was situated at 12:00 - 1:00 o’clock region. Left ventricular and above aortic valve angiography indicated that the VSD location, shape and size, the split vent size on the left ventricle side and its distance from the aortic valve could be correctly measured when the VSD shunt was visualized , which were very helpful in guiding the operator to select the suitable occluder as well as to adjust the release pattern of the occluder. Postoperative imaging findings of the left ventricular and above aortic valve angiography were compared with the preoperative ones. Results Successful occlusion of VSD was obtained in 22 patients , in 13 among them the left ventricular angiography showed that the direction of blood flow beam at the defect hole was from the left ventricle to the right ventricle in an obliquely upward direction. The basal width of the defect on the left ventricle side was (5.12 ± 1.38) mm, and(6 - 10) mm occluder was employed. In the remaining 9 patients the left ventricular angiography showed that the direction of blood flow beam at the defect hole was from the left ventricle to the right ventricle in a direction almost parallel to the aortic valve , and the basal width of the defect on the left ventricle side was (7.18 ± 1.26) mm, and (9 - 12) mm zero-bias occluder was adopted. Interventional occlusion of VSD was unsuccessful in 9 cases as the intercristal hole was rather larger, and two of them had coexisting aortic sinus aneurysm complicated by mid-to-severe degree aortic valve regurgitation. Conclusion Based on the precise analysis of angiographic images by experienced radiologists optimal treatment scheme can be worked out. If conditions permit, symmetrical occluder should be employed so far as possible in order to reduce the degree of operation difficulty and improve the surgical safety and the success rate as well.
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Objective To study the endoscopic anatomical characteristics of the upper petroclival and suprasellar region via subtemproal keyhole approach and to explore the clinical feasibility of this approach to these regions.Methods The operation of subtemporal keyhole approach was performed bilaterally in ten adult cadaver heads fixed with formalin and injected with colored emulsion.The anatomical structures in the upper petroclival region under the endoscope and microscope were recorded.Results Without removal of the zygomatic arch,most of structures in upper petroclival region and suprasellar region were observed under endoscope,including bilateral side anatomical structures; and most perforating branch arteries could be clearly displayed.The natural intracephalic spaces,including posterior communicating artery (PcoA)-oculomotor nerve,posterior communicating artery-anterior choroidal artery (AchA) were important spaces.The stripping of petrous apex and posterior clinoid process could be performed to obtain the ideal surgical exposure.The endoscopic structures in upper petroclival region and suprasellar region may be localized by combination of different markers; the bony structure,such as orifice of internal auditory canal,could be applied as markers.Conclusion The endoscope-assisted neurosurgery through subtemporal keyhole approach is very helpful in exposing the lesions of the upper petroclival and suprasellar region,which minimizes the trauma and is useful in practice.
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<p><b>OBJECTIVE</b>To explore the role of the P38 signaling pathway in the apoptosis of arsenic trioxide (As2 O3)-induced androgen-independent prostate cancer PC-3 cells.</p><p><b>METHODS</b>Androgen-independent prostate cancer PC-3 cells were treated with different concentrations of As2 O3 for 24, 48 and 72 hours. The inhibitory effect of As2 O3 on the cell growth was measured by MTT, the expression of p- P38 detected by Western blot, and the rate of cell apoptosis determined by Annexin V and PI double staining before and after interfering the P38 signaling pathway by SB203580, a highly selective P38 inhibitor.</p><p><b>RESULTS</b>As2 O3 inhibited the proliferation of PC-3 cells in a concentration- and time-dependent manner, and quickly activated P38 phosphorylation, thus giving full play to its biological activities. After 24 hours of treatment with As2 O3 at the concentrations of 2, 10 and 20 micromol/L, the apoptosis rates of the PC-3 cells were (18.9 +/- 0.43), (24.7 +/- 0.29) and (49.7 +/- 1.79)%, respectively, which were reduced to (14.8 +/- 0.81), (22.1 +/- 0.51) and (39.6 +/- 1.74)% after interfering the P38 pathway with SB203580. Inhibition of the P38 pathway significantly reduced the apoptosis of the PC-3 cells induced by As2 O3 (P < 0.05).</p><p><b>CONCLUSION</b>As2 O3 can induce the apoptosis of prostate cancer PC-3 cells by activating the P38 signaling pathway, and interfering the P38 signaling pathway can reduce their apoptosis, which suggests that the P38 signaling pathway is involved in the apoptosis of As2 O3-induced androgen-independent prostate cancer PC-3 cells.</p>
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Humanos , Masculino , Arsenicais , Farmacologia , Linhagem Celular Tumoral , Sistema de Sinalização das MAP Quinases , Óxidos , Farmacologia , Neoplasias da Próstata , Patologia , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno , MetabolismoRESUMO
<p><b>OBJECTIVE</b>Pituitary prolactinoma with severe erectile dysfunction (ED) as the initial symptom is often misdiagnosed. This article explores the diagnosis and treatment of severe ED caused by pituitary prolactinoma.</p><p><b>METHODS</b>We retrospectively analyzed the diagnosis and treatment of 4 cases of pituitary prolactinoma with severe ED (IIEF-5 score 5 - 7) as the initial clinical symptom confirmed by MRI.</p><p><b>RESULTS</b>The 4 cases of pituitary prolactinoma-induced severe ED, with serum prolactin 10 times above the maximum normal level, were misdiagnosed for 2 years. All failed to respond to the PDE5 inhibitor therapy, and then 3 of them underwent transnasal hypophysectomy. Twenty-four months of follow-up found the level of prolactin restored to normal in 1 case (IIEF-5 = 19), and reduced to 600 and 768 IU/L respectively (IIEF-5 = 15) in the other 2. Then administration of the PDE5 inhibitor was followed, which produced satisfactory efficacy. One case was treated with oral bromocriptine, which restored the prolactin level to normal at 12 months (IIEF-5 > 21).</p><p><b>CONCLUSION</b>Prolactin detection and brain MRI can help to confirm pituitary prolactinoma with severe ED at the onset. As for its treatment, in case of an extremely high level of prolactin, simple administration of the PDE5 inhibitor is ineffective. When the prolactin level is reduced after surgery or medication, the symptom of ED can be improved and, in case of no obvious relief, administration of the PDE5 inhibitor can be followed, which may achieve satisfactory results.</p>
Assuntos
Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Disfunção Erétil , Diagnóstico , Inibidores da Fosfodiesterase 5 , Usos Terapêuticos , Neoplasias Hipofisárias , Diagnóstico , Tratamento Farmacológico , Prolactinoma , Diagnóstico , Tratamento Farmacológico , Estudos RetrospectivosRESUMO
<p><b>BACKGROUND</b>Identification of potential serum biomarkers of osteosarcoma to aid in its early diagnosis and in the discovery of possible therapeutic targets is an area of increasing interest.</p><p><b>METHODS</b>Two-dimensional difference-in-gel electrophoresis was used to assess multiple serum samples in patients with osteosarcoma. In addition, differential expression of protein biomarkers was characterized in osteosarcoma serum by using matrix-assisted desorption/ionization time-of-flight mass spectrometry coupled with database interrogation. Serum samples from four individuals with osteosarcoma and four age- and sex-matched healthy control subjects were compared.</p><p><b>RESULTS</b>Fifty-eight significant protein spot features in the osteosarcoma sera were found. These spot features were excised, digested with trypsin, and analyzed with mass spectrometry. Gelsolin was down-regulated only in osteosarcoma. Furthermore, Western blotting and enzyme linked immunosorbent assay (ELISA) confirmed decreased levels of gelsolin in the osteosarcoma serum samples.</p><p><b>CONCLUSIONS</b>These results indicated that gelsolin might have great potential as a biomarker of osteosarcoma and as a potential target for gene therapy.</p>