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Acta Physiologica Sinica ; (6): 225-236, 2022.
Article in Chinese | WPRIM | ID: wpr-927598


This study was to investigate the changes of autophagy in pancreatic tissue cells from hyperlipidemic acute pancreatitis (HLAP) rats and the molecular mechanism of autophagy to induce inflammatory injury in pancreatic tissue cells. Male Sprague Dawley (SD) rats were intraperitoneally injected with caerulein to establish acute pancreatitis (AP) model and then given a high fat diet to further prepare HLAP model. The HLAP rats were treated with autophagy inducer rapamycin or inhibitor 3-methyladenine. Pancreatic acinar (AR42J) cells were treated with caerulein to establish HLAP cell model. The HLAP cell model were treated with rapamycin or transfected with vascular endothelial growth factor (VEGF) siRNA. The inflammatory factors in serum and cell culture supernatant were detected by ELISA method. The histopathological changes of pancreatic tissue were observed by HE staining. The changes of ultrastructure and autophagy in pancreatic tissue were observed by electron microscopy. The expression levels of Beclin-1, microtubule- associated protein light chain 3-II (LC3-II), mammalian target of rapamycin complex 1 (mTORC1), and VEGF were measured by immunohistochemistry and Western blot. The results showed that, compared with control group, the autophagy levels and inflammatory injury of pancreatic tissue cells from HLAP model rats were obviously increased, and these changes were aggravated by rapamycin treatment, but alleviated by 3-methyladenine treatment. In HLAP cell model, rapamycin aggravated the autophagy levels and inflammatory injury, whereas VEGF siRNA transfection increased mTORC1 protein expression, thus alleviating the autophagy and inflammatory injury of HLAP cell model. These results suggest that VEGF-induced autophagy plays a key role in HLAP pancreatic tissue cell injury, and interference with VEGF-mTORC1 pathway can reduce the autophagy levels and alleviate the inflammatory injury. The present study provides a new target for prevention and treatment of HLAP.

Animals , Male , Rats , Acute Disease , Autophagy , Ceruletide/adverse effects , Mammals/metabolism , Mechanistic Target of Rapamycin Complex 1 , Microtubule-Associated Proteins/metabolism , Pancreatitis , RNA, Small Interfering/genetics , Rats, Sprague-Dawley , Sirolimus/adverse effects , Vascular Endothelial Growth Factor A/genetics
Chinese Medical Journal ; (24): 447-455, 2022.
Article in English | WPRIM | ID: wpr-927548


BACKGROUND@#Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and the mechanism of SLE is yet to be fully elucidated. The aim of this study was to explore the role of two-pore segment channel 2 (TPCN2) in SLE pathogenesis.@*METHODS@#Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of TPCN2 in SLE. We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell. Knockdown of TPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting. Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation, apoptosis, and cell cycle of TPCN2-deficient cells. In addition, gene expression profile of TPCN2-deficient cells was analyzed by RNA sequencing (RNA-seq).@*RESULTS@#TPCN2 knockdown with short hairpin RNA (shRNA)-mediated lentiviruses inhibited cell proliferation, and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells. We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells, and screened the differential genes, which were enriched for the G2/M checkpoint, complement, and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways, as well as changes in levels of forkhead box O, phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin, and T cell receptor pathways; moreover, TPCN2 significantly influenced cellular processes and biological regulation.@*CONCLUSION@#TPCN2 might be a potential protective factor against SLE.

Humans , Apoptosis/genetics , Cell Division , Jurkat Cells , Lupus Erythematosus, Systemic/genetics , RNA, Small Interfering/genetics
Article in Chinese | WPRIM | ID: wpr-939687


OBJECTIVE@#To investigate the potential inhibitory effect of interference with PD-L1 on B cell lymphoma in mice.@*METHODS@#Three shRNA vectors for mouse CD274 (PD-L1) were constructed and transiently transfected into 293T cells. RT-qPCR was used to validate the interference efficiency of CD274. The shRNA vector that interfere efficiently with CD274 expression was packaged by using lentivirus packaging system to generate shRNA lentivirus, and then transfected into A20 lymphoma cell line. The methyl thiazol terazolium (MTT) assay was used to detect proliferation after 48 h culture of CD274-sh A20 cells. Meanwhile, BALB/c mice were hypodermically infected with CD274-sh A20 cells. Infected mice were observed daily and assessed to visualize tumor by in vivo fluorescence imaging.@*RESULTS@#The proliferation rate of CD274-sh A20 cells in vitro was significantly lower than that of A20 cells (P<0.05). The tumor size detected by in vivo fluorescence imaging showed a significant reduce in tumor bearing mice with CD274-sh compared with other tumor bearing mice. And the weight and size of tumor in CD274-sh group were also significantly reduced compared with other group (P<0.05). Moreover, the survival time of tumor bearing mice in CD274-sh group was longer than that of the PD-L1 high expression group.@*CONCLUSION@#PD-L1 plays an important role in the incidence and the progression of lymphoma, and the shRNA-based PD-L1 knockdown can inhibit cell proliferation of A20 cells and partly suppress tumor growth.

Animals , Humans , Mice , B7-H1 Antigen/metabolism , Cell Line, Tumor , Lymphoma , Lymphoma, B-Cell , Mice, Inbred BALB C , RNA, Small Interfering/genetics
Chinese Journal of Hepatology ; (12): 38-44, 2022.
Article in Chinese | WPRIM | ID: wpr-935906


Objective: To investigate the effect of adenovirus-mediated shRNA down-regulating phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expression on vinculin, filamin A, and cortactin in activated hepatic stellate cells (HSCs). Methods: Activated rats hepatic stellate cell line (HSC-T6) was cultured in vitro. Recombinant adenovirus Ad-shRNA/PTEN carrying PTEN targeted RNA interference sequence [short hairpin RNA (shRNA)] and empty control virus Ad-GFP were transfected into HSCs. The PTEN mRNA and protein expression of HSCs in each group were detected by real-time fluorescence quantitative PCR and Western blot. The expressional change of vinculin, filamin A and cortactin in HSCs of each group were detected by confocal laser scanning immunofluorescence microscope. Image-pro plus 6.0 software was used for image analysis and processing. The integrated optical density (IOD) of the fluorescence protein expression was measured. The experiment was divided into three groups: control group (DMEM instead of adenovirus solution in the adenovirus transfection step), Ad-GFP group (transfected with empty virus Ad-GFP only expressing green fluorescent protein), and Ad-shRNA/PTEN group (recombinant adenovirus Ad-shRNA/PTEN carrying shRNA targeting PTEN and expressing green fluorescent protein). One-way analysis of variance was used for comparison of mean value among the three groups, and LSD-test was used for comparison between the groups. Results: shRNA targeted PTEN was successfully transfected and the expression of PTEN mRNA and protein in HSC (P < 0.05) was significantly down-regulated. HSCs vinculin was mainly expressed in the cytoplasm. HSCs vinculin fluorescence IOD in the Ad-shRNA/PTEN group (19 758.83 ± 1 520.60) was higher than control (7 737.16 ± 279.93) and Ad-GFP group (7 725.50 ± 373.03) (P < 0.05), but there was no statistically significant difference between control group and Ad-GFP group (P > 0.05). There was no statistically significant difference in the fluorescence IOD of Filamin A among the three groups (P > 0.05), but the subcellular distribution of Filamin A among the three groups were changed. Filamin A in the Ad-shrNA /PTEN HSC group was mainly distributed in the cytoplasm. Filamin A HSC was mainly located in the nucleus.The filamin A HSC in the control group and Ad-GFP group was mainly located in the nucleus. The nucleocytoplasmic ratio of Filamin A in the AD-shrNA /PTEN group (0.60 ± 0.15) was significantly lower than control group (1.20 ± 0.15) and Ad-GFP group (1.08 ± 0.23), P < 0.05. but there was no statistically significant difference in filamin A nucleocytoplasmic ratio of HSC between the control group and the Ad-GFP group (P > 0.05). Cortactin HSCs in the three groups was mainly distributed in the cytoplasm. The cortactin fluorescence IOD of HSCs in the Ad-shRNA/PTEN group was significantly higher than control group (22 959.94 ± 1 710.42) and the Ad-GFP group (22 547.11 ± 1 588.72 ) (P < 0.05), while there was no statistically significant difference in the IOD of cortactin fluorescence in HSCs between the control group and the Ad-GFP group (P > 0.05). Conclusion: The down-regulation of PTEN expression raises the expression of microfilament-binding protein vinculin and cortactin, and changes the subcellular distribution of another microfilament binding protein filamin A, that is, translocation from nucleus to the cytoplasm in activated HSC in vitro.

Animals , Rats , Adenoviridae/metabolism , Carrier Proteins , Cell Proliferation , Cortactin , Filamins/genetics , Hepatic Stellate Cells/metabolism , PTEN Phosphohydrolase/metabolism , RNA, Small Interfering/genetics , Vinculin/genetics
Braz. j. med. biol. res ; 54(4): e10117, 2021. tab, graf
Article in English | LILACS | ID: biblio-1153531


The long noncoding RNA (lncRNA) H19 is involved in the pathogenesis of endometriosis by modulating the proliferation and invasion of ectopic endometrial cells in vitro, but related in vivo studies are rare. This study aimed to investigate the role of lncRNA H19 in a nude mouse model of endometriosis. Ectopic endometrial stromal cells (ecESCs) were isolated from ectopic endometrium of patients with endometriosis and infected with lentiviruses expressing short hairpin RNA (shRNA) negative control (LV-NC-shRNA) or lncRNA-H19 shRNA (LV-H19-shRNA). The ecESCs infected with LV-NC-shRNA and LV-H19-shRNA were subcutaneously implanted into forty 6- to 8-week-old female nude mice. The size and weight of the endometriotic implants were measured at 1, 2, 3, and 4 weeks after implantation and compared, and lncRNA H19 levels in endometriotic implants were evaluated using real-time polymerase chain reaction (RT-PCR). All nude mice survived the experimental period, and no significant differences in body weight were observed between the experimental group and the control group. All nude mice developed histologically confirmed subcutaneous endometriotic lesions with glandular structures and stroma after 1 week of implantation. The subcutaneous lesions in the LV-NC-shRNA group after 1, 2, 3, and 4 weeks of implantation were larger than those in the LV-H19-shRNA group, and lncRNA H19 levels in subcutaneous lesions in the LV-NC-shRNA group were significantly higher than those in the LV-H19-shRNA group. Knockdown of lncRNA H19 suppresses endometriosis in vivo. Further study is required to explore the underlying mechanism in the future.

Humans , Animals , Female , Rabbits , Endometriosis/genetics , RNA, Long Noncoding/genetics , RNA, Small Interfering/genetics , Cell Proliferation/genetics , Endometrium , Mice, Nude
Article in Chinese | WPRIM | ID: wpr-921944


OBJECTIVE@#To investigate the effect of the exosomes from bone marrow mesenchymal stem cells (BMSCs) transfected with silence plasmid of Piezol small interference RNA (siRNA)on osteoarthritis (OA) animal model.@*METHODS@#Twenty male SD rats with specific pathogen free (SPF) were selected, ranging in age from 5.46 to 6.96 months, with a mean of (6.21± 0.75) months;and ranging in weight from 385.76 g to 428.66 g, with a mean of (407.21±21.45) g. BMSCs were extracted. The siRNA silencing plasmid of piezo1 was constructed by siRNA technology. After lentivirus was transfected into BMSCs, the exosomes were extracted. At the cellular level, BMSCs were divided into blank plasmid group and siRNA silencing plasmid group according to whether siRNA-Piezo1 was transfected or not. The osteogenic induction ability of siRNA-Piezo1 on BMSCs was detected by RT-PCR and Western blot. At the animal model level, the OA model was established by surgical resection of cruciate ligament of knee joint.According to different treatment schemes, SD rats were divided into 4 groups:blank control group, model group, BMSCs group and exosome group. SD rats in the blank control group were not treated. In the model group, the cruciate ligaments of rats were excised and OA animal model was established. In BMSCs group, BMSCs were injected into knee joint under CT guidance after OA model establishment, and the cell volume was 5×10@*RESULTS@#The lentivirus transfection efficiency was(92.11±4.22)%. RT-PCR showed that the relative expression of Piezo1 mRNA in blank plasmid group was 1.07±0.06, which was significantly different from that of 0.31±0.01 in siRNA silencing plasmid group (@*CONCLUSION@#Piezo1 siRNA silencing vector can promote the differentiation of BMSCs into chondrocytes and effectively inhibit the progression of OA, so as to delay the disease of OA.

Animals , Male , Rats , Chondrocytes , Disease Models, Animal , Exosomes/genetics , Mesenchymal Stem Cells , Osteoarthritis/therapy , RNA, Small Interfering/genetics , Rats, Sprague-Dawley , Tomography, X-Ray Computed
Article in Chinese | WPRIM | ID: wpr-921928


OBJECTIVE@#To explore the effects of siRNA hsa-circ-0000885 modified bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation, cell proliferation and apoptosis in order to provide new ideas and methods for the clinical treatment of osteoporosis (OP).@*METHODS@#From September 2018 to February 2020, 13 patients with osteoporosis admitted to our hospital were selected as the research objects, including 11 females and 2 males, with an age of (65.45±10.77) years old. After obtaining the informed consent of patients, peripheral blood tissues were extracted. Then the expression level of cir-cRNA in peripheral blood mononuclear cells(PBMC) was detected by circ RNA chip. The expression of circ RNA was silenced by siRNA technology. The BMSCs were transfected with lentivirus. According to the siRNA interference plasmid hsa-circ-0000885, the cells were divided into the blank group, the empty vector group and the siRNA interference group. After 72 hours of treatment, the cell cycle was detected by flow cytometry, the apoptosis level was detected by AV-PI kit, and the osteogenic differentiation ability of BMSCs was detected by ALP staining.@*RESULTS@#The expression of hsa-circ-0000885 in PBMC of patients with osteoporosis was significantly higher than that of healthy controls (@*CONCLUSION@#The lentivirus mediated siRNA hsa-circ-0000885 plasmid transfected into BMSCs and osteoclast co culture system can promote cell proliferation, inhibit apoptosis and promote osteogenic differentiation of BMSCs, which can be used as a potential therapeutic target for OP patients.

Aged , Female , Humans , Middle Aged , Apoptosis/genetics , Cell Differentiation , Cell Proliferation/genetics , Cells, Cultured , Coculture Techniques , Lentivirus , Leukocytes, Mononuclear , Mesenchymal Stem Cells , Osteoclasts , Osteogenesis/genetics , RNA, Small Interfering/genetics , Transfection
Chinese Journal of Biotechnology ; (12): 1237-1248, 2021.
Article in Chinese | WPRIM | ID: wpr-878627


RNA interference (RNAi) is one of the important mechanisms to regulate gene expression in eukaryotes. One of the original functions of RNAi is to facilitate the antiviral strategy of host. Early studies reveal that invertebrates can use RNAi to resist viruses. However, if this mechanism exists in mammals is still controversial. The latest studies confirm that mammals do have the RNAi-based immunity, and researchers believe that RNAi-based antiviral immunity is a brand-new immunological mechanism that was neglected in the past. It is worthy to note that virus can also use RNAi to enhance its infectivity and immune escape in host cells. This review introduces the research history of RNAi-based antiviral immunity in animals and summarizes the main findings in this field. Last but not least, we indicate a series of unresolved questions about RNAi-based antiviral immunity, and explore the relationship between RNAi-based antiviral immunity and other innate immunological pathways. The virus-mediated RNAi pathway in animal is not only an interesting basic biology question, but also has important guiding roles in the development of antiviral drugs.

Animals , Antiviral Agents , Immunity, Innate/genetics , Mammals , RNA Interference , RNA, Small Interfering/genetics , RNA, Viral
Int. braz. j. urol ; 46(6): 950-961, Nov.-Dec. 2020. graf
Article in English | LILACS | ID: biblio-1134248


ABSTRACT Objective To evaluate the effects of Arf6 downregulation on human prostate cancer cells. Materials and Methods The effects of Arf6 downregulation on cell proliferation, migration, invasion and apoptosis were assessed by MTT, BrdU, scratch, Transwell assays and flow cytometry respectively. AKT, p-AKT, ERK1/2, p-ERK1/2 and Rac1 protein expressions were detected by Western blot. Results Downregulating Arf6 by siRNA interference suppressed the mRNA and protein expressions of Arf6. The proliferation capacities of siRNA group at 48h, 72h, and 96h were significantly lower than those of control group (P <0.05). The migration distance of siRNA group at 18h was significantly shorter than that of control group (P <0.01). The number of cells penetrating Transwell chamber membrane significantly decreased in siRNA group compared with that of control group (P <0.01). After 24h, negative control and normal control groups had similar apoptotic rates (P >0.05) which were both significantly lower than that of siRNA group (P <0.01). After Arf6 expression was downregulated, p-ERK1/2 and Rac1 protein expressions were significantly lower than those of control group (P <0.05). Conclusion Downregulating Arf6 expression can inhibit the proliferation, migration and invasion of prostate cancer cells in vitro, which may be related to ERK1/2 phosphorylation and Rac1 downregulation.

Humans , Male , Prostatic Neoplasms/genetics , Down-Regulation , Gene Expression Regulation, Neoplastic , Cell Movement , Apoptosis , RNA, Small Interfering/genetics , Cell Line, Tumor , Cell Proliferation , Neoplasm Invasiveness
Article in Chinese | WPRIM | ID: wpr-880799


OBJECTIVE@#To explore the inhibitory effects of silencing long non-coding RNA (LncRNA) HIF1A-AS2 on epithelialmesenchymal transition (EMT) and tumor stem cell-like phenotype in cervical cancer cells.@*METHODS@#We designed 3 shRNA constructs for silencing HIF1A-AS2 in CaSki cells, and the shRNA with the strongest interference effect was selected for subsequent experiment. CaSki cells were transfected with shRNA-NC or Sh-HIF1A-AS2, and the changes in cell viability, invasion ability, EMT, expressions of EMT-related proteins, formation of cell spheres and expressions of stem cell markers were detected.@*RESULTS@#Transfection with shRNA-NC and Sh-HIF1A-AS2 did not significantly affected the viability of CaSki cells (@*CONCLUSIONS@#Silencing HIF1A-AS2 can inhibit proliferation, invasion and migration of cervical cancer cells

Female , Humans , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Epithelial-Mesenchymal Transition/genetics , Gene Expression Regulation, Neoplastic , Hypoxia-Inducible Factor 1, alpha Subunit , RNA, Long Noncoding/genetics , RNA, Small Interfering/genetics , Uterine Cervical Neoplasms/genetics
Article in Chinese | WPRIM | ID: wpr-880778


OBJECTIVE@#To study the expression of BIRC6 in renal cancer tissues and investigate the effect of BIRC6 silencing on apoptosis and autophagy of 786-O cells.@*METHODS@#Twenty surgical specimens of renal cancer tissues and adjacent renal tissues were collected from Meizhou People's Hospital between February, 2016 and December, 2018 for detection of BIRC6 protein expression using immunohistochemistry. Renal cancer 786-O cells were transfected with a control small interfering RNA (siRNA) or BIRC6 siRNA @*RESULTS@#The expression of BIRC6 protein was significantly higher in renal cancer tissues than in the adjacent renal tissues. Western blotting showed that siRNA-mediated silencing of BIRC6 significantly lowered the expression of BIRC6 in 786-O cells. In the cells with BIRC6 silencing, treatment with 12.5, 25, 50, 100 and 200 μg/mL 5-FU resulted in significantly higher proliferation inhibition rates than in the cells transfected with the control siRNA (@*CONCLUSIONS@#Interference of BIRC6 mediated by siRNA can inhibit autophagy and promote 5-FU-induced apoptosis to enhance the sensitivity of 786-O cells to 5-FU.

Humans , Apoptosis , Autophagy , Cell Line, Tumor , Cell Proliferation , Inhibitor of Apoptosis Proteins/genetics , Kidney Neoplasms/genetics , RNA, Small Interfering/genetics
Chinese Medical Journal ; (24): 2910-2918, 2020.
Article in English | WPRIM | ID: wpr-877921


BACKGROUND@#Psoriasis is a common chronic inflammatory skin disease with 2% to 3% prevalence worldwide and a heavy social-psychological burden for patients and their families. As the exact pathogenesis of psoriasis is still unknown, the current treatment is far from satisfactory. Thus, there is an urgent need to find a more effective therapy for this disease. Keratin 17 (K17), a type I intermediate filament, is overexpressed in the psoriatic epidermis and plays a critical pathogenic role by stimulating T cells in psoriasis. Therefore, we hypothesized that inhibiting K17 may be a potential therapeutic approach for psoriasis. This study aimed to investigate the therapeutic effect of K17-specific small interfering RNA (siRNA) on mice with imiquimod (IMQ)-induced psoriasis-like dermatitis.@*METHODS@#Eight-week-old female BALB/c mice were administered a 5% IMQ cream on both ears to produce psoriatic dermatitis. On day 3, K17 siRNA was mixed with an emulsion matrix and applied topically to the left ears of the mice after IMQ application every day for 7 days. The right ears of the mice were treated in parallel with negative control (NC) siRNA. Inflammation was evaluated by gross ear thickness, histopathology, the infiltration of inflammatory cells (CD3+ T cells and neutrophils) using immunofluorescence, and the expression of cytokine production using real-time quantitative polymerase chain reaction. The obtained data were statistically evaluated by unpaired t-tests and a one-way analysis of variance.@*RESULTS@#The severity of IMQ-induced dermatitis on K17 siRNA-treated mice ears was significantly lower than that on NC siRNA-treated mice ears, as evidenced by the alleviated ear inflammation phenotype, including decreased ear thickness, infiltration of inflammatory cells (CD3+ T cells and neutrophils), and inflammatory cytokine/chemokine expression levels (interleukin 17 [IL-17], IL-22, IL-23, C-X-C motif chemokine ligand 1, and C-C motif chemokine ligand 20) (P < 0.05 vs. the Blank or NC siRNA groups). Compared to the NC siRNA treatment, the K17 siRNA treatment resulted in increased K1 and K10 expression, which are characteristic of keratinocyte differentiation (vs. NC siRNA, K17 siRNA1 group: K1, t = 4.782, P = 0.0050; K10, t = 3.365, P = 0.0120; K17 siRNA2 group: K1, t = 4.104, P = 0.0093; K10, t = 4.168, P = 0.0042; siRNA Mix group: K1, t = 3.065, P = 0.0221; K10, t = 10.83, P < 0.0001), and decreased K16 expression, which is characteristic of keratinocyte proliferation (vs. NC siRNA, K17 siRNA1 group: t = 4.156, P = 0.0043; K17 siRNA2 group: t = 2.834, P = 0.0253; siRNA Mix group: t = 2.734, P = 0.0250).@*CONCLUSIONS@#Inhibition of K17 expression by its specific siRNA significantly alleviated inflammation in mice with IMQ-induced psoriasis-like dermatitis. Thus, gene therapy targeting K17 may be a potential treatment approach for psoriasis.

Animals , Female , Humans , Mice , Dermatitis , Disease Models, Animal , Imiquimod , Inflammation , Keratin-17/genetics , Mice, Inbred BALB C , Psoriasis/genetics , RNA, Small Interfering/genetics , Skin
Braz. j. microbiol ; 48(3): 566-569, July-Sept. 2017. tab, graf
Article in English | LILACS | ID: biblio-889146


Abstract The aim of this study was to assess the in vitro and in vivo effects of short-interfering RNAs (siRNAs) against rabies virus phosphoprotein (P) mRNA in a post-infection treatment for rabies as an extension of a previous report (Braz J Microbiol. 2013 Nov 15;44(3):879-82). To this end, rabies virus strain RABV-4005 (related to the Desmodus rotundus vampire bat) were used to inoculate BHK-21 cells and mice, and the transfection with each of the siRNAs was made with Lipofectamine-2000™. In vitro results showed that siRNA 360 was able to inhibit the replication of strain RABV-4005 with a 1 log decrease in virus titter and 5.16-fold reduction in P mRNA, 24 h post-inoculation when compared to non-treated cells. In vivo, siRNA 360 was able to induce partial protection, but with no significant difference when compared to non-treated mice. These results indicate that, despite the need for improvement for in vivo applications, P mRNA might be a target for an RNAi-based treatment for rabies.

Animals , Phosphoproteins/genetics , Rabies/veterinary , Rabies virus/genetics , Viral Proteins/genetics , Chiroptera/virology , RNA, Small Interfering/genetics , RNA Interference , Phosphoproteins/metabolism , Rabies/virology , Rabies virus/physiology , Viral Proteins/metabolism , Virus Replication , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/metabolism
Rev. chil. pediatr ; 86(4): 287-290, ago. 2015. ilus, graf
Article in Spanish | LILACS | ID: lil-764087


Introducción: La telorragia es un síntoma poco frecuente en pacientes pediátricos, la causa más frecuente en esta población es la ectasia ductal mamaria (EDM), que es una afección benigna y autolimitada, caracterizada por la dilatación del conducto mamario, fibrosis e inflamación periductal. Objetivo: Presentar un caso de EDM, para facilitar el rápido reconocimiento por parte de los médicos, y evitar estudios y tratamientos agresivos. Caso clínico: Lactante de sexo masculino de 6 meses de edad, sano, alimentado por lactancia materna exclusiva; consultó por un nódulo retroareolar derecho y telorragia unilateral. Se realizó una ecografía Doppler que mostró una lesión multiquística, sugerente de una EDM. Se planteó tratamiento expectante y acudió a control a los 6 meses con excelente evolución. Conclusiones: La EDM es la principal causa de telorragia en niños, corresponde a una afección benigna, y la resolución generalmente es espontánea, antes de los 9 meses. Por lo que su conocimiento es de gran relevancia para el adecuado diagnóstico y manejo de estos pacientes.

Introduction: Bloody nipple discharge is an infrequent symptom during childhood. The most common cause in this population is mammary duct ectasia (MDE), which is a benign and self-limiting condition, that is characterized by dilatation of the mammary ducts, fibrosis and periductal inflammation. Objective: Report of a case of MDE in order to improve physicians’ diagnosis accuracy and avoid aggressive studies and treatments. Case report: Six-months old male healthy infant, exclusively breastfeeded, that visited our clinic with a lump beneath his right nipple and bloody discharge from the same nipple. An ultrasound was performed which showed a multicystic lesion suggestive of MDE. Watchful waiting was decided as treatment, with good evolution after six months of follow up. Conclusions: The MDE is the leading cause of bloody discharge in pediatric population, being a benign condition that resolves spontaneously before nine months. The knowledge of this condition is essential so as to accurately diagnose and treat it.

Humans , Cations/chemistry , Indicators and Reagents/chemistry , Lipids/chemistry , Polyenes/chemistry , RNA, Small Interfering/chemistry , Cell Line, Tumor , Chemistry, Pharmaceutical/methods , Gene Transfer Techniques , Genetic Vectors/genetics , HeLa Cells , Liposomes/chemistry , Luciferases/chemistry , Phospholipids/chemistry , RNA, Small Interfering/genetics , Transfection/methods
Arch. argent. pediatr ; 113(3): e137-e139, jun. 2015.
Article in Spanish | LILACS | ID: lil-750470


El síndrome de Wiskott-Aldrich es una inmunodeficiencia primaria; con una incidencia de 3,5 a 5,2 por cada millón de recién nacidos masculinos. Se caracteriza por tener un patrón de herencia recesiva ligada al cromosoma X. En estos pacientes; se ha descrito la tríada clásica de inmunodeficiencia; microtrombocitopenia y eczema. Presentamos un paciente de 5 años de edad; hispánico; con antecedentes de numerosas infecciones desde el primer año de vida. Actualmente; presenta desnutrición crónica; talla baja secundaria y retraso en el desarrollo del lenguaje. Se diagnosticó una mutación poco frecuente del gen asociado al síndrome de Wiskott-Aldrich.

The Wiskott-Aldrich syndrome is a rare X-linked recessive immunodeficiency, with an estimated incidence of 3.5 to 5.2 cases per million males. It is characterizedby immunodeficiency, microthrombocytopenia and eczema. We present a 5-year-old Hispanic male, with a medical history of numerous infectious diseases, compromised health, chronic malnutrition, language delay and failure to thrive. An infrequent mutation in the Wiskott-Aldrich syndrome gene was found.

Animals , Chick Embryo , Avian Proteins/metabolism , Cadherins/metabolism , Neural Stem Cells/cytology , Neural Stem Cells/metabolism , Avian Proteins/antagonists & inhibitors , Avian Proteins/genetics , Base Sequence , Cell Count , Cadherins/antagonists & inhibitors , Cadherins/genetics , Gene Expression Regulation, Developmental , Gene Knockdown Techniques , Neural Tube/cytology , Neural Tube/embryology , Neural Tube/metabolism , Oligonucleotide Array Sequence Analysis , Phenotype , RNA Interference , RNA, Small Interfering/genetics , Signal Transduction
J. appl. oral sci ; 23(2): 179-186, Mar-Apr/2015. graf
Article in English | LILACS, BBO | ID: lil-746537


Since the beginning of their lives, all living organisms are exposed to the influence of geomagnetic fields. Objectives : With respect to the positive effects that magnetic fields have on human tissues, especially the bactericidal effect, this investigation aimed to assess their influence on the reduction of oral microorganisms. Material and Methods : In order to obtain adequate specimens of dental plaque deposit, microbes such as Streptococcus parasanguinis, Staphylococcus epidermidis, Rhodococcus equi and Candida albicans were isolated from the human mouth. To establish the intensity of microbial growth on the basis of the modified optical density (OD) of agar turbidimetry assay, microbial count and spectrophotometry were applied. The study was carried out with two microbial concentrations (1 and 10 CFU/ml) after periods of incubation of 24 and 48 h and using micromagnets. Results : A positive effect of the magnetic field, resulting in the reduction of dental plaque microbes in vitro, was found. In the first 24 hours of exposure to the magnetic field, the number of all isolated microbes was significantly reduced. The most potent influence of magnets and the most intensified reduction after 24 hours were evident in Candida albicans colonies. The decrease in the influence of magnets on microbes in vitro was also detected. Conclusions : Magnets reduce the number of microbes and might be recommended as a supplement in therapy for reduced periodontal tissues. This is important because periodontal tissues that are in good conditions provide prolonged support to the oral tissues under partial and supradental denture. .

Humans , Female , Cell Survival/physiology , Chromosomal Proteins, Non-Histone/metabolism , Cell Line , Cell Survival/genetics , Chromosomal Proteins, Non-Histone/genetics , DNA Breaks, Double-Stranded , DNA Damage/genetics , Fluorescent Antibody Technique , RNA, Small Interfering/genetics
Rev. bras. enferm ; 68(2): 253-260, Mar-Apr/2015. tab
Article in Portuguese | LILACS, BDENF | ID: lil-752516


RESUMO Objetivo: construir e validar um instrumento para monitorar a qualidade dos registros de enfermagem no Programa de Assistência Domiciliar (PAD) em um hospital universitário. Método: estudo metodológico envolvendo a elaboração de um manual e submetido à validação de conteúdo por seis juízes sob consenso ≥ 80%. A coleta ocorreu em 2012 por meio de questionário contendo: evolução de enfermagem, diagnóstico e prescrição de enfermagem e normas para os registros da equipe de enfermagem preconizadas pelo Conselho Regional de Enfermagem-SP e pela instituição. Os itens do manual foram julgados de acordo com as variáveis - relevância, pertinência, clareza e simplicidade. Resultados: das 39 proposições 100% atingiram consenso ≥ 80% em relevância, pertinência e clareza; 92,3% em simplicidade. Os itens sono/repouso, mobilidade e checagem nas atividades prescritas não atingiram consenso mínimo favorável, sendo aprimorados pelas sugestões dos juízes. Conclusão: acreditamos que o instrumento possibilitará a melhoria dos processos de trabalho no PAD. .

RESUMEN Objetivo: construir y validar un instrumento para monitorear la calidad del registros de enfermería en Programa de Atención Domiciliaria (PAD) de un Hospital Universitario. Metodo: estudio metodológico. Fue construido un manual y sometió a validación de contenido por seis jueces bajo el consenso ≥80%. La recogida currió en 2012, con un cuestionario que contiene: evolución de enfermería, diagnóstico y prescripción de enfermería y normas para los registros del personal de enfermaria estabelecidas por Consejo Regional de Enfermería-SP y por la institución. Los artículos del manual fueran juzgadso conforme las variables relevancia, pertinencia, claridad y sencillez. Resultados: de las 39 proposiciones 100% alcanzó consenso ≥ 80% en la relevancia, pertinencia y claridad; 92,3% en la simplicidad. Los itens sueño/resto, movilidad y verificar las actividades prescritas no alcanzó consenso favorable, siendo mejoradas por las sugerencias de los jueces. Conclusión: creemos que el instrumento permitirá la mejora de los procesos de trabajo en PAD. .

ABSTRACT Objective: to build and validate an instrument aimed at monitoring the quality of nursing records in the Home Care Program (HCP) of a university hospital. Method: methodological study involving the elaboration of a manual, whose content was later submitted to six experts for validation, reaching a ≥ 80% consensus. The data collection process was carried out in 2012 by means of a questionnaire comprised of the following issues: nursing evolution, nursing diagnosis, and nursing prescription, and standards for the nursing team recommended by the Regional Nursing Council of São Paulo and by the assessed institution. Manual items were judged according to the following variables: relevance, pertinence, clarity and simplicity. Results: of the 39 propositions, 100% achieved ≥ 80% agreement in the relevance, pertinence and clarity variables; 92.3% in the simplicity variable. Sleep/rest, Mobility and Check-out variables did not reach a favorable minimum consensus in the prescribed activities and were improved following suggestions from the experts. Conclusion: we believe that the instrument will enable the improvement of the HCP’s work process. .

Humans , Actins/metabolism , Cofilin 1/metabolism , Dysentery, Bacillary/microbiology , Nod1 Signaling Adaptor Protein/metabolism , Phosphoprotein Phosphatases/metabolism , Shigella flexneri/physiology , Actins/chemistry , Blotting, Western , Cells, Cultured , Cofilin 1/genetics , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique, Indirect , Gene Expression Regulation , HeLa Cells , High-Throughput Screening Assays , Immunoenzyme Techniques , Immunoprecipitation , Inflammation , Inflammation Mediators/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Nod1 Signaling Adaptor Protein/antagonists & inhibitors , Nod1 Signaling Adaptor Protein/genetics , Phosphorylation , Phosphoprotein Phosphatases/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/genetics , RNA, Small Interfering/genetics , Signal Transduction
Cad. saúde pública ; 31(3): 477-486, 03/2015.
Article in English | LILACS | ID: lil-744826


This paper offers a critical overview of social science research presented at the 2014 International AIDS Conference in Melbourne, Australia. In an era of major biomedical advance, the political nature of HIV remains of fundamental importance. No new development can be rolled out successfully without taking into account its social and political context, and consequences. Four main themes ran throughout the conference track on social and political research, law, policy and human rights: first, the importance of work with socially vulnerable groups, now increasingly referred to as "key populations"; second, continued recognition that actions and programs need to be tailored locally and contextually; third, the need for an urgent response to a rapidly growing epidemic of HIV among young people; and fourth, the negative effects of the growing criminalization of minority sexualities and people living with HIV. Lack of stress on human rights and community participation is resulting in poorer policy globally. A new research agenda is needed to respond to these challenges.

Este artigo oferece uma perspectiva crítica da pesquisa em ciências sociais apresentada na Confe-rência Internacional de AIDS de Melbourne, Aus-trália, em 2014. Em tempos de grandes avanços no campo biomédico, a natureza política do HIV permanece de importância fundamental. Nenhuma inovação será bem-sucedida na prática se desconsiderar o contexto sociopolítico e suas consequências. Quatro temas emergiram da Conferência nos campos do direito, dos direitos humanos e da pesquisa social e política: (1) a importância do trabalho com grupos socialmente vulneráveis, crescentemente chamado de "populações chaves"; (2) o reconhecimento de que ações e programas devem ser sob medida para cada local e contexto; (3) a urgência da resposta a uma epidemia crescendo muito rapidamente entre adolescentes; (4) o efeito negativo da crescente criminalização de minorias sexuais e pessoas vivendo com HIV. Globalmente, a falta de ênfase nos direitos humanos e da participação comunitária tem como resultado políticas públicas de pior qualidade. Precisamos de uma nova agenda de pesquisa para responder a esses desafios.

El artículo ofrece una perspectiva crítica de la investigación en ciencias sociales, presentada en la Conferencia Internacional de SIDA en Melbourne (Australia), 2014. En tiempos de enormes avances biomédicos, la naturaleza política del VIH sigue siendo muy importante. Ninguna innovación será exitosa sin considerar el contexto sociopolítico y sus consecuencias. Cuatro temas surgieron de la conferencia en el campo legal y derechos humanos, además de investigación social y política: (1) la importancia del trabajo con grupos socialmente vulnerables, crecientemente denominados "poblaciones claves"; (2) el reconocimiento de que las acciones y programas deben ser adaptados a un contexto local; (3) la urgencia de una respuesta a una epidemia con crecimiento rápido entre adolescentes; (4) el efecto negativo de la creciente criminalización de las minorías sexuales y personas viviendo con VIH. Globalmente, un limitado énfasis en los derechos humanos y la participación comunitaria tiene como consecuencia peores políticas públicas. Necesitamos una nueva agenda de investigación para responder a estos desafíos.

Animals , Humans , Mice , Gene Silencing/physiology , Huntington Disease/therapy , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Blotting, Northern , Blotting, Western , Cell Line , Gene Expression Regulation/genetics , Huntington Disease/genetics , Huntington Disease/metabolism , Immunohistochemistry , MicroRNAs/genetics , MicroRNAs/physiology , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Plasmids , Polymerase Chain Reaction , RNA, Small Interfering/genetics , RNA, Small Interfering/physiology
Clinics ; 70(2): 120-125, 2/2015. tab, graf
Article in English | LILACS | ID: lil-741425


OBJECTIVES: To explore the microendoscopic discectomy technique and inclusion criteria for the treatment of recurrent lumbar disc herniation and to supply feasible criteria and technical notes to avoid complications and to increase the therapeutic effect. METHODS: A consecutive series of 25 patients who underwent posterior microendoscopic discectomy for recurrent lumbar disc herniation were included. The inclusion criteria were as follows: no severe pain in the lumbar region, no lumbar instability observed by flexion-extension radiography and no intervertebral discitis or endplate damage observed by magnetic resonance imaging. All patients were diagnosed by clinical manifestations and imaging examinations. RESULTS: Follow-up visits were carried out in all cases. Complications, such as nerve injuries, were not observed. The follow-up outcomes were graded using the MacNab criteria. A grade of excellent was given to 12 patients, good to 12 patients and fair to 1 patient. A grade of excellent or good occurred in 96% of cases. One patient relapsed 3 months after surgery and then underwent lumbar interbody fusion and inner fixation. The numerical rating scale of preoperative leg pain was 7.4± 1.5, whereas it decreased to 2.1±0.8 at 7 days after surgery. The preoperative Oswestry disability index of lumbar function was 57.5±10.0, whereas it was 26.0±8.5 at 7 days after surgery. CONCLUSION: In these cases, microendoscopic discectomy was able to achieve satisfactory clinical results. Furthermore, it has advantages over other methods because of its smaller incision, reduced bleeding and more efficient recovery. .

Humans , Centrifugation/methods , Leukocytes, Mononuclear/metabolism , Transfection/methods , Cell Survival/physiology , Leukocytes, Mononuclear/cytology , RNA Interference/physiology , RNA, Small Interfering/genetics
Rev. méd. Chile ; 143(2): 223-236, feb. 2015. ilus, tab
Article in Spanish | LILACS | ID: lil-742574


Prostate cancer represents the second cancer-related cause of death in North American and Chilean men. The main treatment for incurable stages of disease is surgical or pharmacological castration. However, with time and despite the addition of anti-androgens, the disease progresses to a clinical state that has been commonly referred to as “hormone refractory”. In recent years, the concept of hormone refractoriness has been challenged and replaced by “castration resistance”, acknowledging that further and optimal hormonal manipulation can be attained, beyond achieving testosterone levels at castration range. The purpose of this review is to summarize the recent therapeutic breakthroughs in the management of metastatic castrate resistant prostate cancer (mCRPC), with greater emphasis in the newer hormonal therapy agents such as Abiraterone and Enzalutamide. Future combination and sequential treatment strategies are contextualized in the current era of personalized cancer medicine and genomic characterization of prostate cancer.

Animals , Rats , Angiotensin II/physiology , Fibronectins/biosynthesis , Mesangial Cells/metabolism , Plasminogen Activator Inhibitor 1/biosynthesis , Poly(ADP-ribose) Polymerases/physiology , Cells, Cultured , Fibronectins/genetics , Gene Expression Regulation, Enzymologic , Glomerular Mesangium/cytology , Glomerular Mesangium/metabolism , Glomerular Mesangium/pathology , Glomerulonephritis/genetics , Glomerulonephritis/metabolism , Glomerulonephritis/pathology , Mesangial Cells/enzymology , Mesangial Cells/pathology , Plasminogen Activator Inhibitor 1/genetics , Poly(ADP-ribose) Polymerases/biosynthesis , Poly(ADP-ribose) Polymerases/genetics , RNA, Small Interfering/genetics , RNA, Small Interfering/pharmacology