RESUMO
Patients with Parkinson's disease (PD) often suffer from delayed gastric emptying, but the underlying mechanism remains unclear. We have shown previously that a PD rat model comprising bilateral substantia nigra destruction by 6-hydroxydopamine (6-OHDA rats) exhibits gastroparesis with alteration of neural nitric oxide synthase (nNOS) and acetylcholine in gastric corpus. However, changes in pyloric motility in the 6-OHDA rats have not been characterized. Solid gastric emptying test, immunofluorescence, Western blot, and in vitro pyloric motility recordings were used to assess pyloric motor function in the 6-OHDA rats. The 6-OHDA-treated rats displayed delayed solid gastric emptying and a lower basal pyloric motility index. In the 6-OHDA rats, high K+-induced transient contractions were weaker in pyloric sphincters. Electric field stimulation (EFS)-induced pyloric sphincter relaxation was lower in the 6-OHDA rats. NG-nitro-l-arginine methyl ester (l-NAME), a nonselective inhibitor of NOS, markedly inhibited the EFS-induced relaxation in both control and 6-OHDA rats. Pretreatment of tetrodotoxin abolished the effect of EFS on the pyloric motility. In addition, nNOS-positive neurons were extensively distributed in the pyloric myenteric plexus, whereas the number of nNOS-immunoreactive neurons and the protein expression of nNOS were significantly decreased in the pyloric muscularis of 6-OHDA rats. However, sodium nitroprusside-induced pyloric relaxations were similar between the control and 6-OHDA rats. These results indicate that the pyloric sphincters of 6-OHDA rats exhibit both weakened contraction and relaxation. The latter may be due to reduced nNOS in the pyloric myenteric plexus. The dysfunction of the pyloric sphincter might be involved in the delayed gastric emptying.NEW & NOTEWORTHY Reduced nitrergic neurons in pyloric myenteric plexus potently contributed to the attenuated relaxation in 6-hydroxydopamine (6-OHDA) rats, subsequently affecting gastric emptying. SNP could well improve the relaxation of pylori in 6-OHDA rats. The present study provides new insight into the diagnosis and treatment of delayed gastric emptying in patients with PD.
Assuntos
Gastroparesia , Doença de Parkinson , Animais , Gastroparesia/etiologia , Humanos , Óxido Nítrico Sintase Tipo I/metabolismo , Oxidopamina , Piloro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Parkinson's disease (PD) is a common neurodegenerative disorder that is often associated with weak tongue motility. However, the link between the degenerated dopaminergic neurons in the substantia nigra (SN) and lingual dysfunction remains unclear. In the present study, we investigated the localization of dopamine receptor 1 (D1) and dopamine receptor 2 (D2) and alternations in their expression in cholinergic motoneurons of the hypoglossal nucleus (HN) using double-label immunofluorescence, Western blotting and semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR) in rats that received microinjections of 6-hydroxydopamine bilaterally into the SN (6-OHDA rats). The results revealed that a large population of choline acetyltransferase immunoreactive (ChAT-IR) neurons was distributed throughout HN and that almost all of the ChAT-IR motoneurons were also D1-IR and D2-IR. Several tyrosine hydroxylase (TH)-IR profiles were observed in a nonuniform pattern near the ChAT-IR, D1-IR or D2-IR somas, suggesting potent dopaminergic innervation. In the 6-OHDA rats, TH immunoreactivity in the SN was significantly decreased, but food residue was increased and treadmill occupancy time was shortened. In the HN, protein expression of TH and D2 was increased, whereas that of ChAT and D1 was decreased. A similar pattern was observed in mRNA levels. The present study suggests that dopamine may modulate the activity of cholinergic neurons via binding with D1 and D2 in the HN. Changes in the expression of ChAT, TH, D1 and D2 in the HN of 6-OHDA rats might be associated with the impaired tongue motility in PD. These findings should be further investigated.
Assuntos
Neurônios Colinérgicos/metabolismo , Nervo Hipoglosso/metabolismo , Neurônios Motores/metabolismo , Doença de Parkinson Secundária/genética , Receptores de Dopamina D1/genética , Receptores de Dopamina D2/genética , Animais , Colina O-Acetiltransferase/genética , Colina O-Acetiltransferase/metabolismo , Neurônios Colinérgicos/patologia , Modelos Animais de Doenças , Regulação da Expressão Gênica , Humanos , Nervo Hipoglosso/patologia , Masculino , Neurônios Motores/patologia , Oxidopamina , Doença de Parkinson Secundária/induzido quimicamente , Doença de Parkinson Secundária/metabolismo , Doença de Parkinson Secundária/patologia , Ratos , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D2/metabolismo , Transdução de Sinais , Substância Negra/metabolismo , Substância Negra/patologia , Língua/inervação , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Activation of the dopamine (DA) D2 receptor inhibits glucose-stimulated insulin secretion in isolated rodent islets in vitro; however, no information is available regarding the cellular localization of DA receptors (DRs, including D1-D5 receptors) in pancreatic islets in situ. We investigate the protein expression and cellular localization of five types of DRs in pancreatic islets by means of Western blotting and double-labeling immunofluorescence in both normal control and alloxan-induced type 1 diabetes model (T1DM) rats. In control rats, D1 immunoreactivity (-IR) was distributed in the core of the islet and co-localized with insulin-IR, D2-IR was peripherally distributed and found only in somatostatin-immunoreactive cells and D5-IR was co-localized with glucagon-IR and pancreatic polypeptide-IR. No IR for either the D3 or D4 receptor was observed in rat islets. The protein level of the D1 receptor was reduced in T1DM rats (D1/D-glyceraldehyde-3-phosphate dehydrogenase [GAPDH], 0.63 ± 0.05 in control rats compared with 0.16 ± 0.03 in T1DM rats, n = 8, P < 0.05) but no significant alteration was detected in the protein expression of either the D2 receptor (D2/GAPDH, 0.48 ± 0.04 compared with 0.43 ± 0.04, n = 8, P = 0.42) or the D5 receptor (D5/GAPDH, 0.50 ± 0.04 compared with 0.47 ± 0.04, n = 8, P = 0.58). The present study is the first clear demonstration of the protein expression and cellular localization of the D1, D2 and D5 receptors in rat pancreatic islets and provides crucial morphological evidence for further investigations of the underlying mechanism regarding the DA regulation of pancreatic endocrine function.
Assuntos
Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/metabolismo , Receptores Dopaminérgicos/metabolismo , Animais , Diabetes Mellitus Tipo 1/metabolismo , Insulina/metabolismo , Secreção de Insulina , Masculino , Transporte Proteico , Ratos Sprague-Dawley , Frações Subcelulares/metabolismoRESUMO
Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR-enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells.
Assuntos
Abietanos/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Telomerase/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Inibidoras de Apoptose/genética , Proteínas Inibidoras de Apoptose/metabolismo , Survivina , Células U937RESUMO
BACKGROUND AND PURPOSE: Dopamine protects the duodenal mucosa. Here we have investigated the source of dopamine in gastric juice and the mechanism underlying the effects of luminal dopamine on duodenal bicarbonate secretion (DBS) in rodents. EXPERIMENTAL APPROACH: Immunofluorescence, UPLC-MS/MS, gastric incubation and perfusion were used to detect gastric-derived dopamine. Immunofluorescence and RT-PCR were used to examine the expression of dopamine receptors in the duodenal mucosa. Real-time pH titration and pHi measurement were performed to investigate DBS. KEY RESULTS: H+ -K+ -ATPase was co-localized with tyrosine hydroxylase and dopamine transporters in gastric parietal cells. Dopamine was increased in in vivo gastric perfusate after intravenous infusion of histamine and in gastric mucosa incubated, in vitro, with bethanechol chloride or tyrosine. D2 receptors were the most abundant dopamine receptors in rat duodenum, mainly distributed on the apical membrane of epithelial cells. Luminal dopamine increased DBS in a concentration-dependent manner, an effect mimicked by a D2 receptor agonist quinpirole and inhibited by the D2 receptor antagonist L741,626, in vivo D2 receptor siRNA and in D2 receptor -/- mice. Dopamine and quinpirole raised the duodenal enterocyte pHi . Quinpirole-evoked DBS and PI3K/Akt activity were inhibited by calcium chelator BAPTA-AM or in D2 receptor-/- mice. CONCLUSION AND IMPLICATIONS: Dopamine in the gastric juice is derived from parietal cells and is secreted along with gastric acid. On arrival in the duodenal lumen, dopamine increased DBS via an apical D2 receptor- and calcium-dependent pathway. Our data provide novel insights into the protective effects of dopamine on the duodenal mucosa.
Assuntos
Bicarbonatos , Dopamina , Animais , Cromatografia Líquida , Duodeno , Suco Gástrico , Camundongos , Fosfatidilinositol 3-Quinases , Quimpirol/farmacologia , Ratos , Receptores Dopaminérgicos , Receptores de Dopamina D1 , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Acute myeloid leukemia (AML) is a malignant hematological disease, originating from hematopoiesis stem cell differentiation obstruction and clonal proliferation. New reagents or biologicals for the treatment of AML are urgently needed, and exosomes have been identified as candidate biomarkers for disease diagnosis and prognosis. This study aimed to investigate the effects of exosomes from bone marrow mesenchymal stem cells (BMSCs) on AML cells as well as the underlying microRNA (miRNA)-mediated mechanisms. METHODS: Exosomes were isolated using a precipitation method, followed by validation using marker protein expression and nanoparticle tracking analysis. Differentially expressed miRNAs were identified by deep RNA sequencing and confirmed by quantitative real-time polymerase chain reaction (qPCR). Cell proliferation was assessed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt method, and cell cycle progression and apoptosis were detected by flow cytometry. Functional gene expression was analyzed by qPCR and Western blotting (WB). Significant differences were determined using Student's t test or analysis of variance. RESULTS: BMSCs-derived exosomes effectively suppressed cell proliferation (both Pâ<â0.0001 at 10 and 20 µg/mL) and cell cycle progression (Pâ<â0.01 at G0-G1 stage), and also significantly enhanced cell apoptosis (Pâ<â0.001) in KG-1a cells. There were 1167 differentially expressed miRNAs obtained from BMSCs-derived exosomes compared with KG-1a cell-derived exosomes (Pâ<â0.05). Knockdown of hsa-miR-124-5p in BMSCs abrogated the effects of BMSCs-derived exosomes in regulating KG-1a such as the change in cell proliferation (both Pâ<â0.0001 vs. normal KG-1a cell [NC] at 48 and 72 h). KG-1a cells treated with BMSCs-derived exosomes suppressed expression of structural maintenance of chromosomes 4 (Pâ<â0.001 vs. NC by qPCR and Pâ<â0.0001 vs. NC by WB), which is associated with the progression of various cancers. This BMSCs-derived exosomes effect was significantly reversed with knockdown of hsa-miR-124-5p (Pâ<â0.0001 vs. NC by WB). CONCLUSIONS: BMSCs-derived exosomes suppress cell proliferation and cycle progression and promote cell apoptosis in KG-1a cells, likely acting through hsa-miR-124-5p. Our study establishes a basis for a BMSCs-derived exosomes-based AML treatment.
Assuntos
Exossomos , Leucemia Mieloide Aguda , Células-Tronco Mesenquimais , MicroRNAs , Apoptose/genética , Proliferação de Células/genética , Exossomos/genética , Humanos , Leucemia Mieloide Aguda/genética , MicroRNAs/genéticaRESUMO
BACKGROUND: Liver resection for hepatocellular carcinoma (HCC) patients with portal vein tumor thrombus (PVTT) offers a chance of cure, although survival is often limited. The actual 3-year survival and its associated prognostic factors have not been reported. METHODS: A nationwide database of HCC patients with PVTT who underwent liver resection with 'curative' intent was analyzed. The clinicopathologic characteristics, the perioperative, and survival outcomes for the actual long-term survivors were compared with the non-long-term survivors (patients who died within 3 years of surgery). Univariable and multivariable regression analyses were performed to identify predictive factors associated with long-term survival outcomes. RESULTS: The study included 1590 patients with an actuarial 3-year survival of 16.6%, while the actual 3-year survival rate was 11.7%. There were 171 patients who survived for at least 3 years after surgery and 1290 who died within 3 years of surgery. Multivariable regression analysis revealed that total bilirubin > 17.1 µmol/l, AFP > 400 ng/ml, types of hepatectomy, extent of PVTT, intraoperative blood loss > 400 ml, tumor diameter > 5 cm, tumor encapsulation, R0 resection, liver cirrhosis, adjuvant TACE, postoperative early recurrence (< 1 year), and recurrence treatments were independent prognostic factors associated with actual long-term survival. CONCLUSION: One in nine HCC patients with PVTT reached the long-term survival milestone of 3 years after resection. Major hepatectomy, controlling intraoperative blood loss, R0 resection, adjuvant TACE, and 'curative' treatment for initial recurrence should be considered for patients to achieve better long-term survival outcomes.
Assuntos
Carcinoma Hepatocelular , Hepatectomia , Neoplasias Hepáticas , Células Neoplásicas Circulantes/patologia , Veia Porta/patologia , Trombose , Sobreviventes de Câncer/estatística & dados numéricos , Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/cirurgia , China/epidemiologia , Feminino , Hepatectomia/efeitos adversos , Hepatectomia/métodos , Hepatectomia/mortalidade , Humanos , Neoplasias Hepáticas/epidemiologia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/cirurgia , Efeitos Adversos de Longa Duração/epidemiologia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/epidemiologia , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Estudos Retrospectivos , Taxa de Sobrevida , Trombose/etiologia , Trombose/cirurgiaRESUMO
AIM: To assess the patency of pancreaticoenterostomy and pancreatic exocrine function after three surgical methods. METHODS: A pig model of pancreatic ductal dilation was made by ligating the main pancreatic duct. After 4 wk ligation, a total of 36 piglets were divided randomly into four groups. The piglets in the control group underwent laparotomy only; the others were treated by three anastomoses: (1) end-to-end pancreaticojejunostomy invagination (EEPJ); (2) end-to-side duct-to-mucosa sutured anastomosis (ESPJ); or (3) binding pancreaticojejunostomy (BPJ). Anastomotic patency was assessed after 8 wk by body weight gain, intrapancreatic ductal pressure, pancreatic exocrine function secretin test, pancreatography, and macroscopic and histologic features of the anastomotic site. RESULTS: The EEPJ group had significantly slower weight gain than the ESPJ and BPJ groups on postoperative weeks 6 and 8 (P < 0.05). The animals in both the ESPJ and BPJ groups had a similar body weight gain. Intrapancreatic ductal pressure was similar in ESPJ and BPJ. However, pressure in EEPJ was significantly higher than that in ESPJ and BPJ (P < 0.05). All three functional parameters, the secretory volume, the flow rate of pancreatic juice, and bicarbonate concentration, were significantly higher in ESPJ and BPJ as compared to EEPJ (P < 0.05). However, the three parameters were similar in ESPJ and BPJ. Pancreatography performed after EEPJ revealed dilation and meandering of the main pancreatic duct, and the anastomotic site exhibited a variable degree of occlusion, and even blockage. Pancreatography of ESPJ and BPJ, however, showed normal ductal patency. Histopathology showed that the intestinal mucosa had fused with that of the pancreatic duct, with a gradual and continuous change from one to the other. For EEPJ, the portion of the pancreatic stump protruding into the jejunal lumen was largely replaced by cicatricial fibrous tissue. CONCLUSION: A mucosa-to-mucosa pancreatico-jejunostomy is the best choice for anastomotic patency when compared with EEPJ. BPJ can effectively maintain anastomotic patency and preserve pancreatic exocrine function as well as ESPJ.
Assuntos
Anastomose Cirúrgica/métodos , Pâncreas Exócrino , Pancreaticojejunostomia/métodos , Período Pós-Operatório , Animais , Peso Corporal , Humanos , Pâncreas Exócrino/anatomia & histologia , Pâncreas Exócrino/metabolismo , Pâncreas Exócrino/cirurgia , Testes de Função Pancreática , Distribuição Aleatória , SuínosRESUMO
The development of targeted tyrosine kinase inhibitors (TKIs) has succeeded in altering the course of chronic myeloid leukemia (CML). However, a number of patients have failed to respond or experienced disease relapse following TKI treatment. Proviral integration site for moloney murine leukemia virus1 (PIM1) is a serine/threonine kinase that participates in regulating apoptosis, cell cycle, signal transduction and transcriptional pathways, which are associated with tumor progression, and poor prognosis. SMI4a is a selective PIM1 kinase inhibitor that inhibits PIM1 kinase activity in vivo and in vitro. The present study aimed to explore the mechanism underlying the antitumor effect of SMI4a in K562 and imatinibresistant K562 (K562/G) cell lines. It was demonstrated that SMI4a inhibited the proliferation of K562 and K562/G cells using a WST8 assay. The Annexin Vpropidium iodide assay demonstrated that SMI4a induced apoptosis of K562 and K562/G cells in a dose, and timedependent manner. Furthermore, Hoechst 33342 staining was used to verify the apoptosis rate. The clone formation assay revealed that SMI4a significantly inhibited the colony formation capacity of K562 and K562/G cells. Western blot analysis demonstrated that SMI4a decreased phosphorylated (p)Ser9glycogen synthase kinase (GSK) 3ß/pGSK3ß and inhibited the translocation of ßcatenin. In addition, the downstream gene expression of apoptosis regulator Bax and poly(ADPribose) polymerase1 was upregulated, and apoptosis regulator Bcl2 and Myc protooncogene protein expression levels were downregulated. Immunofluorescence results demonstrated changes in the expression level of ßcatenin in the plasma and nucleus. The results of the present study suggest that SMI4a is an effective drug to use in combination with current chemotherapeutics for the treatment of imatinib-resistant CML.
Assuntos
Antineoplásicos/farmacologia , Compostos de Benzilideno/farmacologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Proteínas Proto-Oncogênicas c-pim-1/antagonistas & inibidores , Tiazolidinedionas/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Inibidores de Proteínas Quinases/farmacologia , Transporte Proteico , beta Catenina/metabolismoRESUMO
Recent studies have shown that radiofrequency (RF) ablation therapy is a safe, feasible, and effective procedure for hepatic hemangiomas, even huge hepatic hemangiomas. RF ablation has the following advantages in the treatment of hepatic hemangiomas: minimal invasiveness, definite efficacy, high safety, fast recovery, relatively simple operation, and wide applicability. It is necessary to formulate a widely accepted consensus among the experts in China who have extensive expertise and experience in the treatment of hepatic hemangiomas using RF ablation, which is important to standardize the application of RF ablation for the management of hepatic hemangiomas, regarding the selection of patients with suitable indications to receive RF ablation treatment, the technical details of the techniques, therapeutic effect evaluations, management of complications, etc. A final consensus by a Chinese panel of experts who have the expertise of using RF ablation to treat hepatic hemangiomas was reached by means of literature review, comprehensive discussion, and draft approval.
Assuntos
Ablação por Cateter , Hemangioma/cirurgia , Neoplasias Hepáticas/cirurgia , Ablação por Cateter/efeitos adversos , Ablação por Cateter/mortalidade , China , Consenso , Hemangioma/mortalidade , Hemangioma/patologia , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Complicações Pós-Operatórias/etiologia , Fatores de Risco , Resultado do TratamentoRESUMO
OBJECTIVE: To evaluate the feasibility and efficacy of radiofrequency ablation (RFA) therapy combined with suture and ligation surgery for patients with giant hepatic cavernous hemangioma (HCH). METHODS: Between June 2004 and June 2005, a total of 30 patients were treated by RFA therapy after suture and ligation surgery (SL group, n = 15, with 18 liver lesions) or RFA therapy without suture and ligation surgery (non-SL group, n = 15, with 17 liver lesions) under general anesthesia. All patients had obvious symptoms such as abdominal discomfort, pain and swelling. Preoperative diagnosis of HCH was established by means of ultrasonography, helical computed tomography (CT) scans, and magnetic resonance imaging (MRI). The mean maximum diameter of the lesions was 8.8 cm +/- 1.4 cm. All of the 35 lesions were located on the liver surface, in the caudate lobe of the liver, or adjacent to the gallbladder. Seven patients had chronic calculous cholecystitis, 6 common duct stones, 5 thrombocytopenias, and one posthepatitic cirrhosis. Thirteen of the 30 patients had previous laparotomy. Therapeutic efficacy and clinical data of RFA therapy were compared between these two groups. RESULTS: RFA therapy under ultrasound guidance was performed successfully in all the patients. Cholecystectomy was performed simultaneously for gallstones in 7 patients and for abutting of gallbladder from hemangioma in 2 patients. Choledochotomy with T-tube drainage was performed in 6 patients. The mean blood loss, the mean RFA time per lesion and reduction rate of maximum diameter of the lesions 6 months after RFA in the SL group and non-SL group were 88.0 ml +/- 22.4 ml vs. 255.0 ml +/- 71.7 ml (P < 0.001), 23.0 min +/- 7.5 min vs. 53.3 min +/- 16.0 min (P < 0.001), and 61.8% vs. 44.8% (P < 0.001) respectively. No severe complication related to RFA was observed in all patients. At a median follow-up of 12 months (6 approximately 17 months), a complete lesion necrosis was achieved on the contrast-enhanced helical CT scans in both groups. During the follow-up, all of the 15 patients were free of upper abdominal pain in the SL group, and 12 patients were symptom-free and 3 obtained significant amelioration of symptoms in the non-SL group. CONCLUSION: RFA therapy combined with suture and ligation surgery is a feasible, safe, and effective treatment modality for patients with giant HCHs. It can reduce blood loss, shorten RFA therapy time, and increase therapeutic efficacy of RFA. Intraoperative ultrasonography is a useful adjunct for increasing the therapeutic efficacy of RFA and reducing the complications related to RFA.
Assuntos
Ablação por Cateter , Hemangioma Cavernoso/cirurgia , Neoplasias Hepáticas/cirurgia , Adulto , Estudos de Viabilidade , Feminino , Seguimentos , Humanos , Ligadura , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
Zoledronic acid (ZOL), a nitrogencontaining bisphosphonate, is widely used in metastatic bone disease. Previous studies indicate that ZOL has marked antileukemia activity, however, the underlying mechanism of action remains to be elucidated. The present study aimed to explore the mechanism of the antileukemia effect of ZOL in leukemia cells. It was observed that ZOL inhibited the proliferation of HL60 and adriamycinresistant HL60 (HL60/A) cells using a WST8 assay. An Annexin Vpropidium iodide indicated that ZOL induced apoptosis of the two cell types in a dose and timedependent manner. Hoechst 33342 staining was also used to verify the levels of apoptosis. The colony formation assay demonstrated that ZOL significantly inhibited colony formation capacity in acute myeloid leukemia (AML) cells. This was achieved by the induction of Sphase cell cycle arrest, downregulation of Bcell lymphoma 2 (Bcl2) and upregulation of Bcl2 associated X protein and cleaved poly (ADPribose) polymerase. The results indicate that ZOL inhibited cell proliferation by inducing apoptosis via the mitochondrial apoptotic pathway and this antileukemic activity appeared notably enhanced in HL60/A cells. As ZOL is already available for clinical use, these results indicate that it may be an effective addition to the chemotherapeutic strategies for AML.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Doxorrubicina/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Imidazóis/farmacologia , Apoptose/efeitos dos fármacos , Biomarcadores , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HL-60 , Humanos , Leucemia Mieloide Aguda , Ácido ZoledrônicoRESUMO
The interaction between bone marrow stromal cells and leukemia cells is critical for the persistence and progression of leukemia, and this interaction may account for residual disease. However, the link between leukemia cells and their environment is still poorly understood. In our study, runtrelated transcription factor 3 (RUNX3) was identified as a novel target gene affected by As2O3 and involved in mesenchymal stem cell (MSC)mediated protection of leukemia cells from As2O3induced apoptosis. We observed induction of RUNX3 expression and the translocation of RUNX3 into the nucleus after As2O3 treatment in leukemia cells. In K562 chronic myeloid leukemia cells, downregulation of endogenous RUNX3 compromised As2O3induced growth inhibition, cell cycle arrest, and apoptosis. In the presence of MSC, As2O3induced expression of RUNX3 was reduced significantly and this reduction was modulated by CXCL12/CXCR4 signaling. Furthermore, overexpression of RUNX3 restored, at least in part, the sensitivity of leukemic cells to As2O3. We conclude that RUNX3 plays an important role in As2O3induced cellular responses and allows cells to overcome MSCmediated drug resistance. Therefore, RUNX3 is a promising target for therapeutic approaches to overcome MSCmediated drug resistance.
Assuntos
Antineoplásicos/farmacologia , Arsenicais/farmacologia , Subunidade alfa 3 de Fator de Ligação ao Core/metabolismo , Resistencia a Medicamentos Antineoplásicos/fisiologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Células-Tronco Mesenquimais , Óxidos/farmacologia , Adulto , Apoptose/efeitos dos fármacos , Trióxido de Arsênio , Western Blotting , Linhagem Celular Tumoral , Células Cultivadas , Técnicas de Cocultura , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/terapia , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Adulto JovemRESUMO
Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, has been demonstrated to exert antitumor activity in a variety of cancer cells. The underlying mechanism involves inhibition of cell cycle progression and induction of apoptosis. Besides, celecoxib has also been found to induce autophagy in some solid tumor cells. The aim of this study was to investigate the effect of celecoxib on cell proliferation in HL-60 human acute leukemia cells and to explore the potential mechanism. HL-60 cells were exposed to various concentrations of celecoxib and cell viability was evaluated by the MTT assay. Apoptosis was analyzed with flow cytometry and the amount of autophagosome was evaluated by LysoTracker probe labelling. The expression of apoptosis- and autophagy-related proteins was assayed by Western blot and LysoSensor probe labelling was used to detect the effect of celecoxib on the lysosomal functions. The results of this study indicated that celecoxib inhibited cell proliferation in a time- and dose-dependent fashion. The flow cytometry analysis showed that celecoxib induced apoptosis at low concentrations and mainly cell necrosis at high concentrations. The Western blot test confirmed the induction of apoptosis by the upregulation of apoptosis-related proteins cleaved caspase-3 and cleaved PARP. Furthermore, this study demonstrated that celecoxib prevented the autophagic flux by inhibiting lysosome function; the fluorescence intensity of the LysoTracker probe and the level of autophagy-related proteins LC3-II and p62 were increased, but the fluorescence intensity of the LysoSensor probe was weakened. These findings show that celecoxib is an autophagy suppresser and has antitumor effects in HL-60 cells by inducing cell apoptosis and necrosis.
Assuntos
Antineoplásicos/farmacologia , Autofagia/efeitos dos fármacos , Celecoxib/farmacologia , Leucemia/patologia , Lisossomos/metabolismo , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cloroquina/farmacologia , Células HL-60 , Humanos , Lisossomos/efeitos dos fármacos , Necrose , Fagossomos/efeitos dos fármacos , Fagossomos/metabolismoRESUMO
OBJECTIVE: To evaluate the feasibility, safety and efficacy of radiofrequency ablation (RFA) therapy in patients with hepatic cavernous hemangioma (HCH) and investigate its optimal operative approach. METHODS: Between March 2001 and June 2004, a total of 68 patients, 18 males and 50 females, age 43.1 (30-64), with 104 HCHs 2.5-11 cm in diameter with the mean size of 5.6 cm, were treated by ultrasound-guided RFA, via percutaneous (n = 19), laparoscopic (n = 29), or open surgical (n = 20) approach. In 7 patients with hepatic lesions larger than 7 cm in diameter, Pringle maneuver was used to occlude the hepatic blood flow during the laparoscopic and open RFA therapy. All patients were followed up with helical computed tomographic (CT) scans and ultrasonography for 19 months (6-36 months). RESULTS: Additional intrahepatic lesions not detected preoperatively were found in 2 patients (with 2 new lesions) via laparoscopy and 3 patients (with 4 new lesions) via celiotomy. All patients were treated with RFA successfully. The mean blood loss in the Pringle group (90.0 ml +/- 22.4 ml) was significantly fewer than that in the non-Pringle group (249 ml +/- 56 ml) (P < 0.01). The mean RFA time per lesion in the Pringle group (29.0 min +/- 7.5 min) was shorter markedly compared to the non-Pringle group (55.4 min +/- 12.4 min) (P < 0.01). In the laparoscopic RFA group, laparoscopic cholecystectomy was performed simultaneously in 15 patients with chronic calculous cholecystitis and in another 2 patients because of tumors abutting the gallbladders, and laparoscopic fenestration with intraperitoneal drainage was performed in 3 patients with simple hepatic cysts. In the open RFA group, cholecystectomy was performed in 5 patients with gallbladder diseases, partial cystectomy was performed in one patient with a hepatic cyst, and choledochotomy was performed in 3 patients with common bile duct stones. Postoperative fever and abnormal serum transaminase (ALT and AST) levels were observed in 29 patients (42.6%). A transient hematuria occurred in one patient after open RFA. No specific complications developed during or after RFA. The follow-up showed a complete lesion necrosis rate of 99% (103/104). One patient showed an incomplete lesion necrosis in the margin of RFA site 6 months after percutaneous RFA therapy and obtained retreatment with percutaneous RFA. CONCLUSION: RFA therapy is a safe, feasible and effective treatment options for patients with HCHs. This procedure can be performed via percutaneous, laparoscopic, or open approach. To prevent the RFA-related complications and to increase the therapeutic efficacy of RFA, the choice of optimal operative approach should be based on the lesion size, number, and location and on the patient's clinical status. Hepatic inflow occlusion by Pringle maneuver during laparoscopic or open RFA therapy can reduce the blood loss and increase the therapeutic efficacy significantly.
Assuntos
Ablação por Cateter , Hemangioma Cavernoso/cirurgia , Neoplasias Hepáticas/cirurgia , Adulto , Ablação por Cateter/métodos , Feminino , Humanos , Laparoscopia , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
AIM: To test the efficacy of gene therapy in rat liver tumor. METHODS: A retroviral vector GCIL12EIL2PN encoding human IL-2 (hIL-2) and mouse IL-12 (mIL-12) fused gene and its packaging cell were constructed. The packaging cell lines contained of IL-2 and/or IL-12 genes were injected intrasplenically to transfect splenocyte at different time. The therapeutic effect, immune function and toxic effect were evaluated. RESULTS: The average survival times of the 4 groups using IL genes at days 1, 3, 5 and 7 after tumor implantation were 53.3+/-3.7, 49.3+/-4.2, 31.0+/-2.1 and 24.3+/-1.4 d respectively in IL-2/IL-12 fused gene group, 25.0+/-2.5, 23.5+/-2.0, 18.3+/-2.4 and 12.0+/-1.8 d respectively in IL-2 gene treatment group, and 39.0+/-4.8, 32.0+/-3.9, 23.0+/-2.5 and 19.4+/-2.1 d respectively in IL-12 gene treatment group (P<0.01, n=10). In the IL-12/IL-2 fused gene treatment group, 30% of rats treated at days 1 and 3 survived more than 60 d and serum mIL-12 and hIL-2 levels were still high at day 3 after treatment. Compared with IL alone, NK cell activity was strongly stimulated by IL-2/IL-12 gene. Microscopy showed that livers were infiltrated by a number of lymphocytes. CONCLUSION: IL-2 and/or IL-12 genes injected directly into spleen increase serum IL-2 and IL-12 levels and enhance the NK cell activity, which may inhibit the liver tumor growth. The therapy of fused gene IL-2/IL-12 is of low toxicity and relatively high NK cell activity. Our data suggest that IL-2/IL-12 fused gene may be a safe and efficient gene therapy for liver tumor. The gene therapy should be administrated as early as possible.
Assuntos
Adjuvantes Imunológicos/genética , Antineoplásicos , Interleucina-12/genética , Interleucina-2/genética , Neoplasias Hepáticas/terapia , Baço , Transfecção , Animais , Fusão Gênica Artificial , Vetores Genéticos , Humanos , Masculino , Camundongos , Ratos , Ratos Wistar , Retroviridae/genéticaRESUMO
OBJECTIVE: To study the inhibitory effect of retroviral packaging cells injected intrasplenically encoding mouse interleukin-12 (mIL-12) and human interleukin-2 (hIL-2) fusion gene on the growth of hepatocellular carcinoma. METHODS: The retroviral vectors encoding mIL-12 gene, hIL-2 gene, and mIL-12 and hIL-2 genes, GCIL12EXPN, GCXEIL2PN, and GCIL12EIL2PN were constructed and then transfected into the retroviral packaging cells PA317 to construct cells PA317-GCIL12EXPN, PA317-GCXEIL2PN, and PA317-GCIL12EIL2PN. Rat hepatocellular carcinoma cells CBRH3 were implanted into the livers of Wistar rats to establish hepatoma animal model. Then the rats were divided into 5 groups to be injected intrasplenically with normal saline one day after the implantation (0.8 ml/rat, group I, n = 10), blank vector PA317-GCXEXPN one day after the implantation (10(7) cells/rat, group II, n = 10), PA317-GCIL12EXPN containing IL-2 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group III, n = 40), PA317-GCXEIL2PN containing mIL-12 gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group IV, n = 40), and PA317-GCIL12EIL2PN containing IL-12-IL-2 fusion gene (1 x 10(7) cells/rat 1, 3, 5, or 7 days after the implantation, group V, n = 40) respectively. The rats surviving longer than 2 months were re-injected with hepatocellular carcinoma cells. The therapeutic effect, immune function and toxic effect were evaluated. CT was conducted on the liver before and after the experiment. Laparotomy was performed 3 and 7 days after treatment to resect some of the carcinoma tissues to undergo pathological examination and OX8 immunohistostaining. Serum mIL-12 and hIL-2 were detected one day before and 3, 7, 30, and 60 days after treatment. RESULTS: The average survival times of the rats treated with IL-12-IL-2 fusion gene at the first, third, fifth and seventh day after tumor implantation were 53.3 +/- 3.7 days, 49.3 +/- 4.2 days, 31.0 +/- 2.1 days, and 24.3 +/- 1.4 days respectively, longer than those treated with IL-2 gene (25.0 +/- 2.5 days, 23.5 +/- 2.0 days, 18.3 +/- 2.4 days, and 12.0 +/- 1.8 days respectively, P < 0.001), and those treated with IL-12 gene (39.0 +/- 4.8 days, 32.0 +/- 3.9 days, 23.0 +/- 2.5 days, and 19.4 +/- 2.1 days respectively, P < 0.001). Long survival (>or= 60 days) rate in the rats treated with IL-12-IL-2 fusion gene on the first and third day was 30%. The serum mIL-12 and hIL-2 levels in these rats remained high on the 60th day after treatment. The pathological study showed that the number of infiltrating lymphocytes in liver tumor tissues was increased in the IL-12-IL-2 fusion gene treatment group. CONCLUSION: The retroviral packaging cell line injected intrasplenically encoding mIL-12 and hIL-2 fusion gene inhibits the growth of hepatocellular carcinoma significantly in rats. The therapeutical efficacy of early administration is superior to that of late one.
Assuntos
Terapia Genética , Interleucina-12/genética , Interleucina-2/genética , Neoplasias Hepáticas Experimentais/terapia , Animais , Carcinoma Hepatocelular/terapia , Vetores Genéticos , Humanos , Injeções , Interleucina-12/uso terapêutico , Interleucina-2/uso terapêutico , Masculino , Camundongos , Transplante de Neoplasias , Ratos , Ratos Wistar , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/uso terapêutico , Retroviridae/genética , Baço , TransfecçãoRESUMO
Felty's syndrome (FS) is characterized by the three conditions of rheumatoid arthritis (RA), neutropenia and splenomegaly, and occurs in few cases of longstanding erosive RA. Discriminating between rare occurrences of autoimmune diseases and malignancies is crucial. The present study describes the case of a 17-year-old female with a two-year history of RA, presenting with an irregular fever, hepatosplenomegaly and enlarged lymph nodes. The antinuclear antibody titer was 1:320, while antibody results for anti-dsDNA, anti-Sm and rheumatoid factor were negative. The clinical presentation was similar to that of lymphoma. However, the fluorodeoxyglucose-positron emission tomography and biopsy examinations of the liver and cervical lymph node did not support the diagnosis of lymphoma. According to the laboratory results and clinical symptoms, the differential diagnosis indicated FS, and immunosuppressive agents were administered. Two weeks later, the patient no longer had a fever, and the transaminase levels were normal, associated with shrinkage of the liver and spleen.
RESUMO
Vesicular monoamine transporter 2 (VMAT2) has been exploited as a biomarker of ß-cell mass in human islets. However, a current report suggested no immunoreactivity of VMAT2 in the ß cells of rat islets. To investigate the cellular localization of VMAT2 in islets further, the pancreatic tissues from monkeys and humans were compared with those of rats and mice. The study was performed using among-species comparisons and a type 1 diabetes model (T1DM) for rats by Western blotting, double-label immunofluorescence, and confocal laser scanning microscopy. We found that VMAT2-immunoreactivity (IR) was distributed peripherally in the islets of rodents, but was widely scattered throughout the islets of primates. Consistent with rodent islets, VMAT2-IR did not exist in insulin (INS)-IR cells but was abundantly present in glucagon (GLU)-IR and pancreatic polypeptide (PP)-IR cells in monkey and human islets. VMAT2-IR had no colocalization with INS-IR in any part of the rat pancreas (head, body, and tail). INS-IR cells were reduced dramatically in T1DM rat islets, but no significant alteration in the proportion of VMAT2-IR cells and GLU-IR cells was observed. Furthermore, a strong colocalization of VMAT2-IR with GLU-IR was distributed in the peripheral regions of diabetic islets. For the first time, the current study demonstrates the presence of VMAT2 in α cells and PP cells but not in ß cells in the islets of monkeys and humans. This study provides convinced morphologic evidence that VMAT2 is not present in ß cells. There needs to be studies for new markers for ß cell mass.
Assuntos
Diabetes Mellitus Tipo 1/metabolismo , Modelos Animais de Doenças , Proteínas Vesiculares de Transporte de Monoamina/metabolismo , Animais , Ilhotas Pancreáticas/metabolismo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
To study movement disorder in Parkinson's disease (PD), an animal model of PD can be created by injecting lipopolysaccharide (LPS) into the substantia nigra of rats. In addition to body movement disorders, patients with PD often experience gastrointestinal (GI) dysfunction, such as gastroparesis. However, the underlying mechanism of these disorders remains unclear. The dorsal motor nucleus of vagus (DMV) is a well-known visceral nucleus that regulates GI function. The present study investigated alterations in DMV neurons and gastric motility in rats with LPS-induced PD (LPS-PD rats). Gastric motility was recorded using a strain gauge force transducer in vivo. The distributions of tyrosine hydroxylase (TH)- and choline acetyltransferase (ChAT)-positive neurons in the DMV were determined using immunofluorescence and confocal laser microscopy. Our results indicated that in LPS-PD rats, the number of neurons in the substantia nigra, including neurons with TH immunoreactivity, was markedly reduced, although glial cell proliferation was clearly observed. However, enhanced TH immunoreactivity and decreased ChAT immunoreactivity were found in the DMV. Furthermore, weakened gastric motility was recorded in anesthetized LPS-PD rats. In conclusion, rats with LPS-induced PD exhibited gastric dysmotility with an alteration in DMV neurons. This PD model may be used to study autonomic nervous system disorders that are often observed in patients with early-stage PD.