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1.
Exp Appl Acarol ; 92(4): 871-883, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38656472

RESUMO

The growing concern about migratory birds potentially spreading ticks due to global warming has become a significant issue. The city of Nantong in this study is situated along the East Asia-Australasian Flyway (EAAF), with numerous wetlands serving as roosting sites for migratory birds. We conducted an investigation of hard ticks and determined the phylogenetic characteristics of tick species in this city. We utilized three different genes for our study: the mitochondrial cytochrome oxidase subunit 1 (COX1) gene, the second internal transcribed spacer (ITS2), and the mitochondrial small subunit rRNA (12 S rRNA) gene. The predominant tick species were Haemaphysalis flava (H. flava) and Haemaphysalis longicornis (H. longicornis). Additionally, specimens of Haemaphysalis campanulata (H. campanulata) and Rhipicephalus sanguineus (R. sanguineus) were collected. The H. flava specimens in this study showed a close genetic relationship with those from inland provinces of China, as well as South Korea and Japan. Furthermore, samples of H. longicornis exhibited a close genetic relationship with those from South Korea, Japan, Australia, and the USA, as well as specific provinces in China. Furthermore, R. sanguineus specimens captured in Nantong showed genetic similarities with specimens from Egypt, Nigeria, and Argentina.


Assuntos
Migração Animal , Aves , Complexo IV da Cadeia de Transporte de Elétrons , Ixodidae , Filogenia , Animais , China , Ixodidae/genética , Ixodidae/classificação , Ixodidae/fisiologia , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/análise , RNA Ribossômico/genética , RNA Ribossômico/análise , Ninfa/crescimento & desenvolvimento , Ninfa/classificação , Ninfa/genética , Ninfa/fisiologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/análise , DNA Espaçador Ribossômico/análise
2.
Front Pediatr ; 12: 1367626, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38481738

RESUMO

The occurrence of simultaneous extralobar pulmonary sequestration, esophageal duplication, and bronchogenic cysts is relatively low. We report the case of a 9-month-old Chinese child who had a right lung cyst, detected in utero and was closely monitored until birth. At age 9 months, contrast-enhanced computed tomography revealed right mediastinal extralobar pulmonary sequestration and two cysts. The patient did not exhibit any abnormalities. However, the parents were concerned about the disease. Following positive psychological counseling to the parents, surgery was the strong desire. Subsequently, successful thoracoscopic surgery was performed, excising the three lesions. No postoperative complications occurred. Postoperative pathology confirmed extralobar pulmonary sequestration syndrome combined with esophageal duplication and bronchogenic cysts. The patient was followed-up at 1 and 12 months postoperatively and recovered well with no abnormal space occupation. In such cases, preoperative imaging examinations should be carefully performed, and intraoperative exploration should correspond to that before surgery to avoid lesion omission.

3.
BMC Pediatr ; 23(1): 460, 2023 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-37704985

RESUMO

PURPOSE: This study aimed to evaluate the feasibility and limitations of thoracoscopic segmentectomy of the basal segment (S10). METHODS: Clinical data of 15 children with congenital lung malformations (CLM) who underwent thoracoscopic segmentectomy of S10 via the inferior pulmonary ligament approach from January to October 2022 were retrospectively analyzed. The demographics, clinical presentation, intraoperative time, blood loss, postoperative events, and follow-up duration were assessed. RESULTS: There were 15 patients in this group (nine males and six females). Age ranges from 4.3 to 96.0 months (median, 7.7 months). Fourteen patients underwent S10 segmentectomy, with one undergoing right S10 segmentectomy and right S6 partial wedge resection. The surgical time was 57-125 min (median, 80 min), intraoperative bleeding volume (5-20 ml; median, 10 ml), postoperative drainage tube indwelling (2-4 d; median, 3 d), and postoperative hospitalization time (4-7 d; median, 5 d). No intraoperative conversions, surgical mortalities, or major complications were observed among these patients. Subcutaneous emphysema appeared in three patients; however, it disappeared following conservative observation without pneumothorax or bronchopleural fistula occurrence. CONCLUSIONS: Thoracoscopic segmentectomy of S10 via the inferior pulmonary ligament approach is technically feasible for treating CLM; however, this surgical approach may have certain limitations for CLM with large cysts.


Assuntos
Doenças Pleurais , Pneumonectomia , Feminino , Masculino , Humanos , Criança , Lactente , Pré-Escolar , Estudos Retrospectivos , Ligamentos , Pulmão/diagnóstico por imagem , Pulmão/cirurgia
4.
Front Pediatr ; 11: 1205155, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37342532

RESUMO

Small-intestinal duplication is a rare congenital developmental anomaly that is mainly single; multiple small-intestinal duplications are rare. Most malformations are located in the ileocecal region. The primary surgical treatment is complete resection of the malformations and adjacent intestinal ducts. However, the ileocecal junction plays an important role in children, and it is difficult to preserve it; multiple intestinal repairs increase the risk of postoperative intestinal fistula, which is a challenge for pediatric surgeons. Herein, we report a case of ileocecal preservation surgery for the treatment of multiple small intestinal duplication malformations near the ileocecal area. The child underwent laparoscopically assisted cyst excision and multiple intestinal repairs and had good postoperative recovery and follow-up.

5.
Front Pediatr ; 11: 1143741, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37009266

RESUMO

Intrathoracic gastric duplication has rarely been reported. A 5-year-old child with gastric duplication located in the left thorax was diagnosed and treated successfully using laparoscopy combined with gastroscopy. Preoperative computed tomography, upper gastrointestinal contrast study, ultrasound, and other imaging methods were insufficient for accurate diagnosis in this case. Laparoscopy combined with gastroscopy is more suitable for the diagnosis and treatment of gastric duplication.

6.
Front Pediatr ; 11: 1115101, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36816375

RESUMO

Background: Congenital diaphragmatic hernia (CDH) is a scarce birth defect. It is called late-presenting CDH when symptoms are found after 1 month of life. The clinical manifestations of late-presenting CDH are diverse, among which the most fatal is the cardiac arrest caused by tension gastrothorax. The disease is rare, can easily lead to death owing to improper emergency treatment. This report illustrates the emergency treatment of late-presenting CDH with tension gastrothorax in three Chinese children. Case reports and management: Three children presented to emergency room with a sudden dyspnea, diagnosed accurately by x-ray or computed tomography. In case 1, the gastric tube could not be inserted at the first attempt, and the child cried incessantly. Cardiac arrest occurred when the gastric tube was re-inserted. After cardiopulmonary resuscitation and placement of a thoracic drainage tube, a large amount of gas and stomach contents were drained. Laparoscopic surgery was performed. The patient died of sepsis. In case 2, the gastric tube could not be inserted at the first attempt; consequently, emergency surgery was considered instead of retrying. After the patient was anesthetized, a gastric tube was successfully placed. Subsequently, a large amount of gas and gastric contents was drained, and thoracoscopic surgery was performed. The patient recovered evenly. In case 3, the gastric tube was successfully inserted at the first attempt; however, the vital signs were unstable due to poor drainage of the gastric tube. We injected 20 ml of iohexol into the stomach tube for angiography and dynamic chest film monitoring. After adjusting the position of the stomach tube, the stomach collapsed completely. Thoracoscopic surgery was performed. The patient recovered evenly. Conclusion: Early diagnosis is essential for children with late-presenting CDH complicated by tension gastrothorax. Fully collapsing the stomach is a key step in emergency treatment. In addition, gastric tube insertion is the first choice. In children with difficulty in gastric tube placement at the first attempt, the gastric tube can be placed under anesthesia, and emergency surgery performed simultaneously. Endoscopic surgery can be the first choice in cases of complete stomach collapse.

7.
Front Pediatr ; 10: 934827, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36061392

RESUMO

Objective: The present study aimed to evaluate the safety and feasibility of modified thoracoscopic wedge resection of limited peripheral lesions in the posterior basal segment (S10) in children with congenital pulmonary airway malformation (CPAM). Materials and methods: We retrospectively analyzed the clinical data of children with CPAM who underwent thoracoscopic modified wedge resection at our institution from November 2020 to February 2022. The surgical method was as follows: we marked the external boundary of the lesion with an electric hook, dissected and retained the segmental vein between the lesion and normal lung tissue as the internal boundary, cut the arteries, veins, and bronchus entering the lesion, and cut and sealed the lung tissue between the internal and external boundaries with LigaSure™ to complete the modified wedge resection. Results: A total of 16 patients were included, aged 3.8-70.0 months and weighing 6.5-21.0 kg. The intraoperative course was uneventful in all patients. The median operation time and intraoperative bleeding volume were 74 min (50-110 min) and 5 mL (5-15 mL), respectively. The median postoperative drainage tube indwelling time was 3 days (2-4 days), and the median postoperative hospital stay was 6 days (4-8 days). Pathological diagnosis included two cases of type 1, 10 cases of type 2, and four cases of type 3 CPAM. There were no cases of intraoperative conversion, surgical mortality, or major complications. However, subcutaneous emphysema occurred in two children, which spontaneously resolved without pneumothorax orbronchopleural fistula development. All patients were followed up for a median period of 10 months (3-18 months), and there were no cases of hemoptysis or residual lesions on chest computed tomography. Conclusion: Modified thoracoscopic wedge resection via the inferior pulmonary ligament approach is safe and feasible for children with CPAM with limited peripheral lesions in S10.

8.
Transl Pediatr ; 11(8): 1408-1414, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36072530

RESUMO

Background: Lung is the most common site of metastasis in pediatric patients with Wilms tumor (WT). For such patients, neoadjuvant chemotherapy before nephrectomy is recommended now. A considerable proportion of metastases will shrink in size after the chemotherapy. However, there are still some of them that are not sensitive to chemotherapy and require subsequent surgical resection. For pediatric patients with bilateral lung metastases from WT which are not sensitive to chemotherapy, the simultaneous surgical removal of bilateral lung tumors via one-stage surgery is problematic. These children typically require 2 separate surgeries to remove the bilateral lung metastases and improve their 5-year event-free survival (EFS) rate. There is no precedent in pediatric thoracic surgery for one-stage, bilateral, lung wedge resection via subxiphoid approach video-assisted thoracic surgery (SA-VATS). Case Description: In this article, we report on a successful SA-VATS performed on an 8-year-old boy whereby all of the bilateral lung metastases were completely resected. The operation was performed through 3 incisions under the xiphoid process and costal arch. No complications occurred after surgery. The patient's intraoperative blood loss was approximately 20 mL. Drainage tubes were indwelled in both pleural cavities which were removed on post-operative day (POD) 5 and POD 6. There was no recurrence at follow-up of about 4 months. Conclusions: This case presents a new option for thoracoscopic surgery which is safe and less invasive for patients with bilateral lung metastases from WT. Similar patients may benefit from the shorter time frame between the operation and other postoperative treatment.

9.
Front Pediatr ; 10: 926942, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35935357

RESUMO

To the best of our knowledge, double or multiple extralobar pulmonary sequestrations (PSs) with anomalous arterial supply in the ipsilateral thoracic cavity have rarely been reported before. PS can be divided into two types: intralobar sequestration (ILS) and extralobar sequestration (ELS). We encountered a 5-month-old infant with double ELS in the left thoracic cavity that was incidentally detected during thoracoscopic surgery. Surgical exploration revealed two separate, well-circumscribed abnormal masses in the left thoracic cavity, and the patient was successfully treated using thoracoscopic surgery. Postoperative pathology confirmed that both masses were PS tissues. Accurate preoperative diagnosis using CT alone may be inadequate in this type of case. Therefore, thoracoscopy may be more suitable for diagnosing and treating unusual ELS.

10.
Front Microbiol ; 11: 407, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32231652

RESUMO

Bacterial glycoproteins have been investigated as vaccine candidates as well as diagnostic biomarkers. However, they are poorly understood in Mycobacterium bovis strain bacille Calmette-Guérin (BCG), a non-pathogenic model of Mycobacterium tuberculosis. To understand the roles of secreted O-mannosylated glycoproteins in BCG, we conducted a ConA lectin-affinity chromatography and mass spectra analysis to identify O-mannosylated proteins in BCG culture filtrate. Subsequent screening of antigens was performed using polyclonal antibodies obtained from a BCG-immunized mouse, with 15 endogenous O-mannosylated proteins eventually identified. Of these, BCG_0470 and BCG_0980 (PstS3) were revealed as the immunodominant antigens. To examine the protective effects of the antigens, recombinant antigens proteins were first expressed in Mycobacterium smegmatis and Escherichia coli, with the purified proteins then used to boost BCG primed-mice. Overall, the treated mice showed a greater delayed-type hypersensitivity response in vivo, as well as stronger Th1 responses, including higher level of IFN-γ, TNF-α, and specific-IgG. Therefore, mannosylated proteins BCG_0470 and BCG_0980 effectively amplified the immune responses induced by BCG in mice. Together, our results suggest that the oligosaccharide chains containing mannose are the antigenic determinants of glycoproteins, providing key insight for future vaccine optimization and design.

11.
Zhejiang Da Xue Xue Bao Yi Xue Ban ; 47(3): 255-260, 2018 05 25.
Artigo em Chinês | MEDLINE | ID: mdl-30226325

RESUMO

OBJECTIVE: To summarize experience in the treatment of complex congenital intestinal atresia in children, so as to investigate the key points and effect of the operation. METHODS: Medical notes of 49 children with complex intestinal atresia treated between January 2012 and January 2018 were reviewed. The information of age, sex, age at operation, full-term or premature, birth weight, clinical manifestation, auxiliary examination, preliminary diagnosis, treatment process, discharge diagnosis, pathological results and prognosis of patients were analyzed. RESULTS: All patients underwent surgical treatment, including 42 cases with laparotomy (85.7%) and 7 with laparoscopic surgery (14.3%); 1 case undergoing laparoscopic surgery was converted to laparotomy due to meconium peritonitis. The mean operation time was (147±43) min (70-270 min); the mean fasting time after surgery was (8±3) d (4-16 d); the mean parenteral nutrition time was (12±6) d (3-30 d). Eleven cases were discharged against medical after operation and lost to follow-up. Among rest 38 children, 1 child (2.6%) received intestinal resection and ostomy five days after operation due to gastrointestinal perforation; 1 child (2.6%) received conservative treatment one month later due to adhered intestinal obstruction and left hospital with cure; 1 child (2.6%) received enterodialysis and ileostomy eight days after operation due to anastomotic leak, and received the operation for the closure of fistula after three months; 4 children had complications including fluid and electrolyte disorders, anemia, hypoproteinemia and so on, and recovered after conservative treatments. Postoperative follow-up showed that 1 child with duodenal atresia had lower body weight at 6 month after operation, but the body weight returned to normal when the child was one year old; 1 child with preterm labor of 32 weeks was treated with enteral nutrition, and gradually restored the normal diet after 6 months. Growth retardation was not observed in other children. CONCLUSIONS: With active treatment and reservation of normal bowel tube as much as possible during the operation, the prognosis of children with complex intestinal atresia is usually favorable.


Assuntos
Anormalidades Congênitas , Atresia Intestinal , Criança , Anormalidades Congênitas/cirurgia , Anormalidades Congênitas/terapia , Obstrução Duodenal/cirurgia , Obstrução Duodenal/terapia , Nutrição Enteral , Humanos , Lactente , Recém-Nascido , Atresia Intestinal/cirurgia , Atresia Intestinal/terapia , Laparoscopia , Estudos Retrospectivos
12.
J Colloid Interface Sci ; 512: 555-566, 2018 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-29100160

RESUMO

Visible-light-driven (VLD) Ag/AgBr/ZnFe2O4 composites with different weight ratios of ZnFe2O4 were synthesized via a facile hydrothermal method. The ZnFe2O4 was evenly dispersed on the surface of Ag/AgBr particles with the diameter about 20 nm. The obtained Ag/AgBr/ZnFe2O4 composites exhibited the high bacterial disinfection efficiency and inactivated bacteria after 120 min visible illumination, which was better than those of with pure ZnFe2O4 and Ag/AgBr. Due to the introduction of ZnFe2O4, the Ag/AgBr/ZnFe2O4 inactivated bacterial cells through the generation of H2O2, which generated from the electron reduction on the conduction band of ZnFe2O4 in the system. The photocatalytic experiments indicated that as-prepared samples showed the good photocatalytic performance toward degradation of methyl orange (MO) dye. In a word, the Ag/AgBr/ZnFe2O4 composites, as the antibacterial photocatalyst, is a promising candidate material in wastewater decontamination.


Assuntos
Brometos/química , Desinfecção/métodos , Escherichia coli/efeitos dos fármacos , Compostos Férricos/química , Luz , Compostos Orgânicos/química , Compostos de Prata/química , Poluentes Químicos da Água/química , Óxido de Zinco/química , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/efeitos da radiação , Processos Fotoquímicos , Prata/química
13.
Int J Mol Sci ; 16(12): 28320-33, 2015 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-26633363

RESUMO

Adenylyl Cyclase 3 (AC3) plays an important role in the olfactory sensation-signaling pathway in mice. AC3 deficiency leads to defects in olfaction. However, it is still unknown whether AC3 deficiency affects gene expression or olfactory signal transduction pathways within the main olfactory epithelium (MOE). In this study, gene microarrays were used to screen differentially expressed genes in MOE from AC3 knockout (AC3(-/-)) and wild-type (AC3(+/+)) mice. The differentially expressed genes identified were subjected to bioinformatic analysis and verified by qRT-PCR. Gene expression in the MOE from AC3(-/-) mice was significantly altered, compared to AC3(+/+) mice. Of the 41266 gene probes, 3379 had greater than 2-fold fold change in expression levels between AC3(-/-) and AC3(+/+) mice, accounting for 8% of the total gene probes. Of these genes, 1391 were up regulated, and 1988 were down regulated, including 425 olfactory receptor genes, 99 genes that are specifically expressed in the immature olfactory neurons, 305 genes that are specifically expressed in the mature olfactory neurons, and 155 genes that are involved in epigenetic regulation. Quantitative RT-PCR verification of the differentially expressed epigenetic regulation related genes, olfactory receptors, ion transporter related genes, neuron development and differentiation related genes, lipid metabolism and membrane protein transport etc. related genes showed that P75NTR, Hinfp, Gadd45b, and Tet3 were significantly up-regulated, while Olfr370, Olfr1414, Olfr1208, Golf, Faim2, Tsg101, Mapk10, Actl6b, H2BE, ATF5, Kirrrel2, OMP, Drd2 etc. were significantly down-regulated. In summary, AC3 may play a role in proximal olfactory signaling and play a role in the regulation of differentially expressed genes in mouse MOE.


Assuntos
Adenilil Ciclases/deficiência , Mucosa Olfatória/metabolismo , Transcriptoma , Animais , Apoptose/genética , Proliferação de Células , Biologia Computacional/métodos , AMP Cíclico/metabolismo , Epigênese Genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Camundongos , Camundongos Knockout , Anotação de Sequência Molecular , Neurônios Receptores Olfatórios/metabolismo , Reprodutibilidade dos Testes , Transdução de Sinais
14.
Oncol Lett ; 10(5): 2957-2961, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26722271

RESUMO

Hepatocellular carcinoma is a lethal malignancy with poor prognosis, partially due to tumor metastasis, recurrence and resistance to chemo- or radio-therapy. Cisplatin can inhibit cancer cell DNA replication, and is widely used in the clinical treatment of tumors. The present study aimed to generate eukaryotic expression vectors for Livin shRNA and to examine the effects of Livin shRNA on the chemosensitivity of HepG2 hepatocellular carcinoma cells. Eukaryotic expression vectors for Livin shRNA (pSD11-U6/Neo/GFP/Livin) were designed and constructed. The HepG2 hepatocellular carcinoma cell line was transfected with this vector using the liposome method. The expression levels of Livin mRNA and protein were measured by quantitative polymerase chain reaction and western blot analysis. The rate of cell growth inhibition was measured using MTT assay following treatment of the cells with cisplatin (2.0 mg/l). DNA sequencing confirmed that the construction of the eukaryotic expression vector for Livin shRNA had been successful. Transfection of these vectors into HepG2 cells led to a significant reduction in the expression levels of Livin mRNA and protein (P<0.05). Cisplatin treatment was associated with significantly higher rates of cell growth inhibition in HepG2 cells transfected with Livin shRNA compared with those that were not transfected (P<0.05). The vectors constructed in the present study produced effective inhibition of the Livin gene in HepG2 cells and increased the chemosensitivity of hepatocellular carcinoma cells.

15.
Asian Pac J Cancer Prev ; 14(9): 4943-52, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24175757

RESUMO

Hepatocellular carcinoma (HCC) is one of the most common malignancies, with a very poor prognosis. Despite significant improvements in diagnosis and treatment in recent years, the long-term therapeutic efficacy is poor, partially due to tumor metastasis, recurrence, and resistance to chemo- or radio-therapy. Recently, it was found that a major feature of tumors is a combination of unrestrained cell proliferation and impaired apoptosis. There are now 8 recognized members of the IAP-family: NAIP, c-IAP1, c-IAP2, XIAP, Survivin, Bruce, Livin and ILP-2. These proteins all contribute to inhibition of apoptosis, and provide new potential avenues of cancer treatment. As a powerful tool to suppress gene expression in mammalian cells, RNAi species for inhibiting IAP genes can be directed against cancers. This review will provide a brief introduction to recent developments of the application IAP-siRNA in tumor studies, with the aim of inspiring future treatment of HCC.


Assuntos
Apoptose , Carcinoma Hepatocelular/terapia , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Terapia Genética , Proteínas Inibidoras de Apoptose/antagonistas & inibidores , Neoplasias Hepáticas/terapia , RNA Interferente Pequeno/uso terapêutico , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteína 3 com Repetições IAP de Baculovírus , Carcinoma Hepatocelular/genética , Humanos , Proteínas Inibidoras de Apoptose/genética , Neoplasias Hepáticas/genética , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Proteína Inibidora de Apoptose Neuronal/antagonistas & inibidores , Proteína Inibidora de Apoptose Neuronal/genética , Survivina , Ubiquitina-Proteína Ligases , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/antagonistas & inibidores , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/genética
16.
Asian Pac J Cancer Prev ; 14(9): 5467-72, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24175843

RESUMO

OBJECTIVE: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin and Survivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors on the biological behavior of HepG2 cells. METHODS: shRNA eukaryotic expression vectors pSD11-Livin and pSD11- Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNA and protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay. RESULTS: The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected into HepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected with pSD11-Livin and pSD11-Survivin were 0.12 ± 0.02 and 0.33 ± 0.13, respectively, which was significantly lower than levels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expression levels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to single- transfection (P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in the single-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to the greatest extent in the co-transfection group relative to any other group (P<0.05). CONCLUSIONS: Co-transfection with pSD11-Livin and pSD11-Survivin was more efficient than transfection with either vector alone in reducing the mRNA and protein expression of Livin and Survivin genes in HepG2 cells. Co-transfection also inhibited the proliferation of transfected cells more than the other groups, and induced cellular apoptosis more effectively.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Apoptose , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos/farmacologia , Proteínas Inibidoras de Apoptose/metabolismo , Proteínas de Neoplasias/metabolismo , RNA Interferente Pequeno/genética , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Western Blotting , Primers do DNA/química , Primers do DNA/genética , Vetores Genéticos/administração & dosagem , Células Hep G2 , Humanos , Proteínas Inibidoras de Apoptose/antagonistas & inibidores , Proteínas Inibidoras de Apoptose/genética , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Survivina , Transfecção
17.
Acta Pharmacol Sin ; 34(4): 507-14, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23524565

RESUMO

AIM: To investigate whether NO over-production in rat mesangial cells cultured in high glucose (HG) is related to activation of the TGF-ß1/PI3K/Akt pathway. METHODS: Rat mesangial cells line (HBZY-1) was exposed to HG (24.44 mmol/L) or H2O2 (10 µmol/L) for 16 h. NO release was quantified using the Griess assay. The TGF-ß1 level was measured using ELISA. The protein expression of p-Akt, t-Akt, Bim, and iNOS was examined by Western blotting. The mRNA levels of TGF-ß1 and Bim were measured using RT-PCR. The cell proliferation rate was estimated using a BrdU incorporation assay. RESULTS: Treatment of the cells with HG, H2O2, or TGF-ß1 (5 ng/mL) significantly increased the NO level that was substantially inhibited by co-treatment with the NADPH oxidase inhibitor diphenylene iodonium (DPI), TGF-ß1 inhibitor SB431542, or PI3K inhibitor LY294002. Both HG and H2O2 significantly increased the protein and mRNA levels of TGF-ß1 in the cells, and HG-induced increases of TGF-ß1 protein and mRNA were blocked by co-treatment with DPI. Furthermore, the treatment with HG or H2O2 significantly increased the expression of phosphorylated Akt and iNOS and cell proliferation rate, which was blocked by co-treatment with DPI, SB431542, or LY294002. Moreover, the treatment with HG or H2O2 significantly inhibited Bim protein and mRNA expression, which was reversed by co-treatment with DPI, SB431542, or LY294002. CONCLUSION: The results demonstrate that high glucose causes oxidative stress and NO over-production in rat mesangial cells in vitro via decreasing Bim and increasing iNOS, which are at least partially mediated by the TGF-ß1/PI3K/Akt pathway.


Assuntos
Glucose/metabolismo , Células Mesangiais/metabolismo , Óxido Nítrico/metabolismo , Estresse Oxidativo/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proteína 11 Semelhante a Bcl-2 , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Peróxido de Hidrogênio/farmacologia , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Células Mesangiais/efeitos dos fármacos , NADPH Oxidases/antagonistas & inibidores , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Óxido Nítrico/genética , Estresse Oxidativo/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/genética , Fosforilação/efeitos dos fármacos , Fosforilação/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , RNA Mensageiro/genética , Ratos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fator de Crescimento Transformador beta1/genética
18.
Exp Physiol ; 98(4): 934-45, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23159718

RESUMO

Oxidative stress has been shown to play an important role in the development and progression of diabetic nephropathy, and the formation of reactive oxygen species (ROS) is a direct consequence of hyperglycaemia. We hypothesized that hyperglycaemia-induced ROS can activate the transforming growth factor-ß1 (TGF-ß1)-phosphoinositide 3-kinase (PI3K)-Akt-FoxO3a signalling pathway, negatively regulating expression of manganese superoxide dismutase (MnSOD), which promotes excessive ROS generation and accelerates the pathological process of diabetic nephropathy. In vitro, in rat mesangial cells, high glucose (30 mmol l(-1)), but not equimolar mannitol, stimulated ROS production, upregulated the levels of TGF-ß1, increased the phosphorylated Akt/total Akt and phosphorylated FoxO3a/total FoxO3a protein ratios, altered the subcellular localization of FoxO3a and reduced the levels of MnSOD expression. These high-glucose-induced changes further promoted the generation of ROS. In vivo, in db/db mice treated with an inhibitor of TGF-ß1 (SB431542) or PI3K (LY294002), the levels of phosphorylated Akt and phosphorylated FoxO3a in the kidney cortices were decreased, the level of MnSOD expression was increased and the level of the lipid peroxidation end-product, malondialdehyde, was reduced. We conclude that overproduction of ROS induced by a high glucose concentration decreases the expression of MnSOD via the PI3K-Akt-FoxO3a pathway and further aggravates oxidative stress in diabetic nephropathy.


Assuntos
Nefropatias Diabéticas/metabolismo , Estresse Oxidativo/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Superóxido Dismutase/metabolismo , Animais , Células Cultivadas , Glucose/metabolismo , Córtex Renal/metabolismo , Células Mesangiais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Ratos
19.
Fitoterapia ; 82(6): 920-6, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21605637

RESUMO

Quercetin's protective effects on the glomerulosclerosis of diabetic nephropathy (DN) in rat mesangial cells were investigated. The cell cycles, type IV collagen and laminin, TGF-ß(1) mRNA, Smad 2/3 and Smad 7, and activities of cell antioxidases were measured. Compared with the high glucose group, quercetin may decrease the cell percentages of G(0)/G(1) phase, Smad 2/3 expression, laminin and type IV collagen, and TGF-ß(1) mRNA level significantly. The antioxidant capacity, the cell percentages of S phase and Smad 7 expression was significantly increased by quercetin. These results suggest that quercetin is a protective agent against glomerulosclerosis in DN.


Assuntos
Antioxidantes/farmacologia , Nefropatias Diabéticas/metabolismo , Matriz Extracelular/efeitos dos fármacos , Células Mesangiais/efeitos dos fármacos , Quercetina/farmacologia , Animais , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Colágeno Tipo IV/efeitos dos fármacos , Colágeno Tipo IV/metabolismo , Matriz Extracelular/metabolismo , Glucose/efeitos adversos , Hipertrofia , Laminina/efeitos dos fármacos , Laminina/metabolismo , Células Mesangiais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , Ratos , Proteína Smad2/efeitos dos fármacos , Proteína Smad2/metabolismo , Proteína Smad3/efeitos dos fármacos , Proteína Smad3/metabolismo , Proteína Smad7/efeitos dos fármacos , Proteína Smad7/metabolismo , Fator de Crescimento Transformador beta1/efeitos dos fármacos , Fator de Crescimento Transformador beta1/genética
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