RESUMO
BACKGROUND: Until now there has been no study that directly compares the effect of lansoprazole and pantoprazole administered intravenously on intragastric acidity. The aim of this study is to compare the effect of lansoprazole (30 mg) and pantoprazole (40 mg) administered intravenously on gastric acidity. MATERIAL/METHODS: Helicobacter pylori-negative healthy volunteers were recruited in this open-label, randomized, two-way crossover, single centre study. Lansoprazole at 30 mg or pantoprazole at 40 mg was intravenously administered twice daily for 5 consecutive days with at least a 14-day washout interval. Twenty-four-hour intragastric pH was continuously monitored on days 1 and 5 of each dosing period. RESULTS: Twenty-five volunteers completed the 2 dosing periods. The mean intragastric pH values were higher in subjects treated with lansoprazole than those with pantoprazole on both day 1 (6.41 ± 0.14 vs. 5.49 ± 0.13, P=0.0003) and day 5 (7.09 ± 0.07 vs. 6.64 ± 0.07, P=0.0002). Significantly higher percentages of time with intragastric pH >4 and pH >6 were found in the subjects treated with lansoprazole than those with pantoprazole on day 1 (pH >4, 87.12 ± 4.55% vs. 62.28 ± 4.15%, P=0.0012; pH >6, 62.12 ± 4.12% vs. 47.25 ± 3.76%, P=0.0216) and pH >6 on day 5 (76.79 ± 3.77% vs. 58.20 ± 3.77%, P=0.0025). CONCLUSIONS: Intravenous lansoprazole produces a longer and more potent inhibitory effect on intragastric acidity than does intravenous pantoprazole.
Assuntos
2-Piridinilmetilsulfinilbenzimidazóis/administração & dosagem , Antiulcerosos/administração & dosagem , Ácido Gástrico , Adulto , Sequência de Bases , China , Estudos Cross-Over , Primers do DNA , Feminino , Humanos , Concentração de Íons de Hidrogênio , Infusões Intravenosas , Lansoprazol , Masculino , Pantoprazol , Reação em Cadeia da Polimerase , Valores de ReferênciaRESUMO
BACKGROUND: Patients diagnosed with extrahepatic bile duct carcinoma (EBDC) have a poor prognosis. OBJECTIVE: The purpose of these studies was to design radioactive stents for EBDC and to evaluate the feasibility and safety of the stents in healthy pigs. DESIGN: Plastic stents with inserted iodine-125 seeds were designed and tested in 11 healthy pigs. The pigs were divided into 4 groups on the basis of radiation doses. INTERVENTIONS: The stents with estimated radiation dose at a 5-mm radial distance from the axis of the seeds of 30 Gy, 60 Gy, and 90 Gy were implanted in the common bile duct (CBD) in groups A, B, and C (n = 3 in each group), with the control group (n = 2) being implanted with the stents containing nonradioactive seeds. MAIN OUTCOME MEASUREMENTS: Histologic evaluation was performed under a light microscope. RESULTS: The procedures were successfully performed on all pigs. Severe hyperplasia of the mucosa was seen in the control group. In the experimental groups, obvious mucosal necrosis near the radioactive seeds was observed but without perforation of the CBD wall. In lower-dose groups (30 Gy), mild hyperplasia of mucosal glands with fibrosis under the necrosis layer was seen. However, after the increase of the dose, mucosal glands were disappearing without a visible mucosal layer. CONCLUSIONS: The radioactive stents are safe at each dose in healthy pigs. Moreover, our observations indicate the feasibility to design specific radioactive stents according to the size, shape, and position of EBDC in future clinical applications.
Assuntos
Neoplasias dos Ductos Biliares/radioterapia , Ductos Biliares Extra-Hepáticos , Braquiterapia/instrumentação , Stents , Animais , Ductos Biliares Extra-Hepáticos/patologia , Desenho de Prótese , Suínos , Fatores de TempoRESUMO
Epigenetic modifications play an important role during carcinogenesis. The main goal of this study was to examine expression levels of two critical enzymes, DNA methyltransferase-1 (DNMT1) and histone deacetylase-1 (HDAC1), by immunohistochemistry (IHC) in human pancreatic cancer and precancerous lesions: 20 foci containing normal ductal epithelial cells without an inflammatory back-ground (DE), 30 containing ductal epithelial cells with an inflammatory background (DEI), 48 of pancreatic intraepithelial neoplasia-1A (PanIN-1A), 103 of PanIN-1B, 99 of PanIN-2, 30 of PanIN-3, 18 of intraductal papillary mucinous neoplasm A (IPMA), 10 of IPMB, 20 of IPMC, and 54 of pancreatic ductal adenocarcinoma (PDAC). The expression levels of both DNMT1 and HDAC1 increased from normal to precancerous lesions to pancreatic cancer, in a malignancy-dependent manner. Correlations between expression levels and clinicopathological features of the 54 PDAC patients were also analyzed. The expression of DNMT1 significantly correlated with nerve infiltration, degree of tumor differentiation and TNM staging (p<0.05), while that of HDAC1 correlated with proliferative activity, degree of tumor differentiation and TNM staging (p<0.05). Patients with higher expression of DNMT1 and/or HDAC1 had an overall lower survival than those with lower expression (p<0.05). Higher expression of DNMT1 and HDAC1 correlated with advanced stages of the disease and reflect the malignancy of pancreatic carcinoma. They may become new prognostic markers and potential therapeutic targets for pancreatic cancer.
Assuntos
Adenocarcinoma Mucinoso/enzimologia , Adenocarcinoma/enzimologia , Carcinoma in Situ/enzimologia , Carcinoma Ductal Pancreático/enzimologia , Carcinoma Papilar/enzimologia , DNA (Citosina-5-)-Metiltransferases/análise , Histona Desacetilases/análise , Neoplasias Pancreáticas/enzimologia , Adenocarcinoma/patologia , Adenocarcinoma Mucinoso/patologia , Carcinoma in Situ/mortalidade , Carcinoma in Situ/patologia , Carcinoma in Situ/terapia , Carcinoma Ductal Pancreático/mortalidade , Carcinoma Ductal Pancreático/patologia , Carcinoma Ductal Pancreático/terapia , Carcinoma Papilar/patologia , Diferenciação Celular , Proliferação de Células , DNA (Citosina-5-)-Metiltransferase 1 , Feminino , Histona Desacetilase 1 , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/terapia , Fatores de Tempo , Resultado do Tratamento , Regulação para CimaRESUMO
PURPOSE: Patients diagnosed with pancreatic cancer typically have a poor prognosis. The aims of these studies were to design radioactive stents and to evaluate the feasibility and safety of the stents in animals. EXPERIMENTAL DESIGN: To combine the effects of stents and brachytherapy, plastic stents with inserted iodine-125 seeds were designed and tested in 18 normal pigs. The pigs were divided into five groups on the basis of radiation dose. The estimated radiation dose at a 5-mm radial distance from the axis of the seeds was 50 Gy in group A, 100 Gy in group B, 150 Gy in group C, and 200 Gy in group D, with four pigs in each group. In the control group (n = 2), the same plastic stents with non-radioactive seeds were implanted in the pancreatic duct. RESULTS: The procedures were successfully done on 14 of 18 (78%) pigs, whereas pancreatic duct perforation occurred in four pigs (22%). The thickened wall of the dilated pancreatic duct was clearly observed in the control group. However, the normal morphologic structure of the pancreatic duct wall disappeared in the experimental groups. Histopathologic examination revealed that the stents were surrounded with necrotic tissues and lateral fibrous tissues. During the follow-up period, the width of outside fibrous tissues gradually increased. CONCLUSIONS: These results indicate that the radioactive stents are safe in all dose groups, and it is feasible to design a special radioactive stent for each patient according to the size, shape, and position of the pancreatic tumor.
Assuntos
Braquiterapia/instrumentação , Terapia Combinada/métodos , Radioisótopos do Iodo/uso terapêutico , Ductos Pancreáticos/cirurgia , Neoplasias Pancreáticas/terapia , Stents , Animais , Braquiterapia/efeitos adversos , Braquiterapia/métodos , Humanos , Plásticos , Prognóstico , Radiometria , Projetos de Pesquisa , Stents/efeitos adversos , Suínos , Fatores de Tempo , Resultado do TratamentoRESUMO
OBJECTIVE: To evaluate the feasibility and safety of radioactive pancreatic duct stents implanted in the pancreatic ducts of pigs by endoscopy. METHODS: Different doses of 125I radioactive pancreatic duct stents were implanted in the pancreatic ducts of pigs by endoscopy. Blood tests were conducted before and after implantation. 14, 30 and 60 days after implantation of the radioactive stents, the pigs were euthanized in batch. All animals underwent post mortem examination to exclude intra-abdominal hemorrhage, pancreatic fistula or peritonitis. During autopsy, the liver, bile ducts, head of the pancreas, stomach and duodenum were examined for perforation, stricture or dilation and damage of the surrounding structures. RESULTS: Fourteen pigs were implanted with pancreatic duct stents by endoscopic procedures. There was no effusion, hemorrhage or necrosis in the adjacent duodenum, stomach, liver or right kidney. The normal morphological structures of the duct of Wirsung disappeared in all the treated pigs. Histopathological examination revealed that the stents were surrounded by necrotic tissue and outside fibrous tissue. During the follow-up period, the width of outside fibrous tissue gradually increased. There were no serious abnormalities noted in the blood tests after implantation. CONCLUSION: It is indicated that the radioactive stents are safe in all the different dose groups. For future clinical application, it is feasible to design a special radioactive stent for each patient according to the size, shape and position of the pancreatic tumor.
Assuntos
Braquiterapia/métodos , Ductos Pancreáticos/efeitos da radiação , Próteses e Implantes , Animais , Cateteres de Demora , Endoscopia , Estudos de Viabilidade , Radioisótopos do Iodo , Neoplasias Pancreáticas/radioterapia , Stents , SuínosRESUMO
AIM: To construct a recombinant live attenuated Salmonella typhimurium DNA vaccine encoding H pylori ureB gene and mouse IL-2 gene and to detect its immunogenicity in vitro and in vivo. METHODS: H pylori ureB and mouse IL-2 gene fragments were amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. DNA sequence of the amplified ureB and IL-2 genes was assayed, then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions resulting in pIRES-ureB and pIRES-ureB-IL-2. The recombinant plasmids were used to transform competent E. coli DH5alpha, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-ureB and pIRES-ureB-IL-2 were used to transform LB5000 and the recombinant plasmids extracted from LB5000 were finally introduced into the final host SL7207. After that, recombinant strains were grown in vitro repeatedly. In order to detect the immunogenicity of the vaccine in vitro, pIRES-ureB and pIRES-ureB-IL-2 were transfected to COS-7 cells using Lipofectamine 2000, the immunogenicity of expressed UreB and IL-2 proteins was assayed with SDS-PAGE and Western blot. C57BL/6 mice were orally immunized with 1 x 10(8) recombinant attenuated Salmonella typhimurium DNA vaccine. Four weeks after vaccination, mice were challenged with 1 x 10(7) CFU of live H pylori SS1. Mice were sacrificed and the stomach was isolated for examination of H pylori 4 wk post-challenge. RESULTS: The 1700 base pair ureB gene fragment amplified from the genomic DNA was consistent with the sequence of H pylori ureB by sequence analysis. The amplified 510 base pair fragment was consistent with the sequence of mouse IL-2 in gene bank. It was confirmed by PCR and restriction enzyme digestion that H pylori ureB and mouse IL-2 genes were inserted into the eukaryotic expression vector pIRES. The experiments in vitro showed that stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying ureB and IL-2 genes was successfully constructed and the specific strips of UreB and IL-2 expressed by recombinant plasmids were detected through Western blot. Study in vivo showed that the positive rate of rapid urease test of the immunized group including ureB and ureB-IL-2 was 37.5% and 12.5% respectively, and was significantly lower than that (100%) in the control group (P<0.01). CONCLUSION: Recombinant attenuated Salmonella typhimurium DNA vaccine expressing UreB protein and IL-2 protein with immunogenicity can be constructed. It can protect mice against H pylori infection, which may help the development of a human-use H pylori DNA vaccine.
Assuntos
Infecções por Helicobacter/prevenção & controle , Helicobacter pylori/genética , Vacinas contra Salmonella/genética , Salmonella typhimurium/genética , Vacinas de DNA/genética , Animais , Proteínas de Bactérias/genética , Células COS , Chlorocebus aethiops , DNA Bacteriano/genética , Infecções por Helicobacter/imunologia , Interleucina-2/genética , Interleucina-2/metabolismo , Camundongos , Infecções por Salmonella/prevenção & controle , Salmonella typhimurium/imunologia , Transfecção , Urease/genética , Vacinas AtenuadasRESUMO
OBJECTIVE: To investigate the mRNA expression of GLI1, a transcription regulator of Hedgehog signaling pathway, in human pancreatic carcinoma and to explore its clinical significance. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of GLI1 in the tumor tissues, tissues near tumor, and normal tissues obtained during operation from 25 pancreatic carcinoma patients. RESULTS: The GLI1 mRNA expression rate of the tumor tissues was 68.0% (17/25), significantly higher than those of the tissues near tumor, and normal tissues [24.0% (6/25) and 0 (0/25) respectively, both P < 0.01]. The GLI1 mRNA expression rate was significantly associated with the differentiation degree of tumor tissue (P = 0.014), and not significantly associated with the tumor size, invasion, and metastasis (all P > 0.05). CONCLUSION: GLI1 mRNA expression is strong in pancreatic carcinoma tissues and is significantly associated with the differentiation degree of tumor tissue. With diagnostic implication, GLI1 mRNA expression may be regarded as a parameter of determining the degree of malignancy and prognosis of pancreatic carcinoma.
Assuntos
Regulação Neoplásica da Expressão Gênica , Neoplasias Pancreáticas/patologia , Fatores de Transcrição/genética , Idoso , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/genética , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína GLI1 em Dedos de ZincoRESUMO
AIM: To examine the effect of canstatin, a newly discovered endogenous inhibitor of angiogenesis, in the treatment of pancreatic cancer in vivo. METHODS: The canstatin cDNA fragment was synthesized and amplified from the total RNA extracted from human placenta tissues by RT-PCR. The resulting product was firstly cloned into pUCm-T vector, then into plasmid pET-22b (+) and transformed into E. coli BL21. Isopropyl-1-thio-b-Dgalactopyran-oside (IPTG) was used to induce the expression of canstatin protein and affinity chromatography was used to purify the protein. To determine the activity of purified recombinant human canstatin (rhCanstatin), orthotopic xenograft human pancreatic cancer models were established. Human pancreatic cancer cells (SW1990) were injected into the pancreas of BALB/c nude mice. Twenty-four nude mice with orthotopic xenograft tumor were randomly divided into 3 groups 10 d after the inoculation, and were treated with PBS 0.3 mL, or canstatin 5 mg/kg, or 10 mg/kg per day for 3 wk intraperitoneally. When the experiment was over, all tumors were resected and the effects of rhCanstatin on tumor growth, microvessel density (MVD) were analyzed. RESULTS: After IPTG induction, SDS-PAGE showed a new monomeric 24 kDa protein band. This protein was purified through affinity chromatography and refolded through dialysis with a final concentration of 60 mg/L. In orthotopic pancreatic cancer models, the final tumor volume in groups treated with PBS, canstatin 5 mg/kg, 10 mg/kg were 355.21+/-39.54 mm3, 112.73+/-10.47 mm3, and 61.75+/-6.99 mm3 respectively. The immunohistochemical examination showed that the MVD in tumors treated with canstatin was significantly less than that in other group. CONCLUSION: These findings demonstrate that the rhCanstatin effectively retards the growth of pancreatic cancer in a dose-dependent manner through inhibiting angiogenesis and may be a promising therapeutic agent for pancreatic cancer treatment in the clinic.
Assuntos
Colágeno Tipo IV/uso terapêutico , Neoplasias Pancreáticas/tratamento farmacológico , Fragmentos de Peptídeos/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células , Colágeno Tipo IV/genética , DNA Complementar/genética , Relação Dose-Resposta a Droga , Escherichia coli/genética , Humanos , Neovascularização Patológica/tratamento farmacológico , Neoplasias Pancreáticas/patologia , Fragmentos de Peptídeos/genética , Plasmídeos/genética , RNA/genética , Proteínas Recombinantes/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: One of the major causes of death in severe acute pancreatitis (SAP) is severe infection owing to bacterial translocation. Some clinical studies suggested that ecoimmunonutrition (EIN) as a new strategy had better treatment effect on SAP patients. But the experiment studies on the precise mechanism of the effect of EIN were less reported. In this study, we mainly investigated the effects of EIN on bacterial translocation in SAP model of dogs. METHODS: SAP was induced by retrograde infusion of 5% sodium taurocholate into the pancreatic duct in healthy hybrid dogs. The SAP dogs were supported with either parenteral nutrition (PN) or elemental enteral nutrition (EEN) or EIN. The levels of serum amylase, serum aminotransferase and plasma endotoxin were detected before and after pancreatitis induction. On the 7th day after nutrition supports, peritoneal fluid, mesenteric lymph nodes (MLN), liver, and pancreas were collected for bacterial culture with standard techniques to observe the incidence of bacterial translocation. Pathology changes of pancreas were analyzed by histopathologic grading and scoring of the severity of pancreas, and the degree of intestinal mucosal damage was assessed by measuring mucosal thickness, villus height, and crypt depth of ileum. RESULTS: Compared with PN and EEN, EIN significantly decreased the levels of serum amylase, serum aminotransferase, plasma endotoxin, and the incidence of bacterial translocation. Furthermore, compared with the others, the histology scores of inflammation in pancreas and the ileum injury (ileum mocosa thickness, villus height, and crypt depth) were significantly alleviated by EIN (P < 0.05). Moreover, concerning liver function, the serum levels of alanine aminotransferase, aspartate aminotransferase and albumin were ameliorating significantly in the EIN group. CONCLUSION: Our results suggested that EIN could maintain the integrity of intestinal mucosal barrier and reducing the incidence of bacterial translocation in SAP dogs. Early EIN was safe and more effective treatment for SAP dogs.
Assuntos
Mucosa Intestinal/metabolismo , Apoio Nutricional , Pancreatite/terapia , Doença Aguda , Amilases/metabolismo , Animais , Bactérias/isolamento & purificação , Cães , Endotoxinas/sangue , Nutrição Enteral , Imunidade , Fígado/fisiopatologia , Pâncreas/patologia , Pancreatite/imunologia , Pancreatite/metabolismo , Nutrição ParenteralRESUMO
AIM: To construct a recombinant attenuated Salmonella typhimurium DNA vaccine carrying Helicobacter pylori hpaA gene and to detect its immunogenicity. METHODS: Genomic DNA of the standard H pylori strain 17 874 was isolated as the template, hpaA gene fragment was amplified by polymerase chain reaction (PCR) and cloned into pUCmT vector. DNA sequence of the amplified hpaA gene was assayed, then cloned into the eukaryotic expression vector pIRES through enzyme digestion and ligation reactions. The recombinant plasmid was used to transform competent Escherichia coli DH5alpha, and the positive clones were screened by PCR and restriction enzyme digestion. Then, the recombinant pIRES-hpaA was used to transform LB5000 and the recombinant plasmid isolated from LB5000 was finally used to transform SL7207. After that, the recombinant strain was grown in vitro repeatedly. In order to identify the immunogenicity of the vaccine in vitro, the recombinant pIRES-hpaA was transfected to COS-7 cells using Lipofectamine2000, the immunogenicity of expressed HpaA protein was detected with SDS-PAGE and Western blot. RESULTS: The 750-base pair hpaA gene fragment was amplified from the genomic DNA and was consistent with the sequence of H pylori hpaA by sequence analysis. It was confirmed by PCR and restriction enzyme digestion that H pylori hpaA gene was inserted into the eukaryotic expression vector pIRES and a stable recombinant live attenuated Salmonella typhimurium DNA vaccine carrying H pylori hpaA gene was successfully constructed and the specific strip of HpaA expressed by pIRES-hpaA was detected through Western blot. CONCLUSION: The recombinant attenuated Salmonella typhimurium DNA vaccine strain expressing HpaA protein with immunogenicity can be constructed and it may be helpful for further investigating the immune action of DNA vaccine in vivo.
Assuntos
Helicobacter pylori/genética , Hemaglutininas/genética , Salmonella typhimurium/genética , Vacinas Atenuadas/genética , Vacinas de DNA/genética , Adesinas Bacterianas , Animais , Vacinas Bacterianas/genética , Células COS , Clonagem Molecular , Helicobacter pylori/imunologia , Hemaglutininas/imunologia , Reação em Cadeia da Polimerase , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Salmonella typhimurium/imunologiaRESUMO
OBJECTIVE: To clone human canstatin gene and express its recombinant protein. METHODS: The total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy-mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG). RESULTS: The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET-22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction. CONCLUSION: Human canstatin gene was successfully cloned and its recombinant proteins were expressed in this study.
Assuntos
Colágeno Tipo IV/biossíntese , Fragmentos de Peptídeos/biossíntese , Proteínas Recombinantes/biossíntese , Sequência de Bases , Clonagem Molecular/métodos , Colágeno Tipo IV/genética , Escherichia coli/genética , Vetores Genéticos , Humanos , Técnicas In Vitro , Fragmentos de Peptídeos/genética , Proteínas Recombinantes/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Urban runoff is an increasingly important source of excess phosphorus (P) to local receiving waters. Bioretention, a promising technology for urban stormwater pollution treatment, was investigated to determine whether the mixture of purple soil and sand could adsorb sufficient P at low concentrations in urban stormwater. The TP concentrations of urban runoff from variously impervious areas in Chongqing City ranged from 0. 04 to 7. 00 mg . L-1 (mean ± S. D. = 0. 75 mg . L-1 ± 1. 08 mg . L-1); the TDP concentrations ranged from 0. 02-0. 46 mg . L-1 ( mean ± S. D. = 0. 15 mg . L-1 ± 0. 10 mg . L-1). The media adsorption benchmark was determined for a bioretention facility sized at 10% of the 100% impervious catchment area and having 10 years of capacity according to annual rainfall pattern and the runoff TDP range. The media benchmark for adsorption was calculated as 7. 5 mg . kg-1 at soluble P concentration of 0. 30 mg . L-1 which provided the necessary stormwater treatment. The oxalate-extractable aluminum and iron content influenced the P sorption capacity for neutral and acid purple soils. A strong positive linear relationship was observed between the oxalate ratio [OR = (Alox + Feox)/Pox] and media P sorption capacity. The media mixture of 20% purple soil and 80% sand showed excellent P removal, meeting the developed benchmark for adsorptive behavior. The media mixture in a large-scale (60 cm) column consistently produced soluble reactive phosphorus effluent event with mean concentrations <0. 05 mg . L-1. The media mixture of purple soil and sand can be used as a bioretention media to treat low-concentration phosphorus in urban runoff under various hydrologic and pollutant concentration conditions.
Assuntos
Fósforo/isolamento & purificação , Movimentos da Água , Poluentes da Água/isolamento & purificação , Adsorção , China , Cidades , Chuva , Dióxido de Silício/química , Solo/químicaRESUMO
AIM: To determine the distribution of cagG gene of Helicobacter pylori (H pylori) isolates cultured from patients with various digestive diseases and its relationship with gastroduodenal diseases. METHODS: cagG was amplified by polymerase chain reaction in 145 H pylori isolates cultured from patients with chronic gastritis (n=72), duodenal ulcer (n=48), gastric ulcer (n=17), or gastric and duodenal ulcer (n=8), and the relationship between cagG status and the grade of gastric mucosal inflammation was determined. RESULTS: cagG was present in 91.7% of the 145 H pylori isolates, with the rates were 90.3%, 93.8%, 88.2% and 100.0%, respectively, in those from patients with chronic gastritis, duodenal ulcer, gastric ulcer, and gastric and duodenal ulcer. There was no significant difference among the four groups (P>0.05). The average grade of gastric mucosal inflammation in the antrum and corpus was 1.819+/-0.325 and 1.768+/-0.312, respectively in cagG positive patients, whereas the average inflammation grade was 1.649+/-0.297, 1.598+/-0.278 respectively in cagG negative cases (P>0.05). CONCLUSION: cagG gene of H pylori was quite conservative, and most H pylori strains in Chinese patients were cagG positive. cagG status was not related to clinical outcome or the degree of gastric mucosal inflammation. Therefore, cagG can not be used as a single marker for discrimination of H pylori strains with respect to a specific digestive disease.
Assuntos
Gastrite/microbiologia , Gastrite/patologia , Infecções por Helicobacter/microbiologia , Infecções por Helicobacter/patologia , Helicobacter pylori/genética , Adenocarcinoma/microbiologia , Adenocarcinoma/patologia , Adolescente , Adulto , Idoso , Povo Asiático , Feminino , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Úlcera Péptica/microbiologia , Úlcera Péptica/patologia , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/patologiaRESUMO
OBJECTIVE: To evaluate the killing effects of the cytosine deaminase (CD) gene mediated by adenovirus vector on human pancreatic cancer cell lines in vitro. METHODS: The CD gene was cloned into pAdTrack-CMV-CD, and pAdTrack-CMV-CD and pAdEasy-1 were recombinated in bacteria. The newly recombinated Ad-CD containing green fluorescent protein (GFP) was propagated in 293 cells and purified by cesium chloride gradient centrifugation. Human pancreatic cancer cell lines Patu8988 and SW1990 were infected with this virus, then 5-FC was added. XTT assay was used to estimate relative numbers of viable cells. RESULTS: The positive clones were selected by using endonuclease to digest the combinatants and the concentration of viral liquids containing the CD gene was 2X10(11) pfu/ml. It was found that significant cytotoxic activities were possessed by 5-FC for the CD gene transduced pancreatic cell lines, but little effects exerted on the nontransduced pancreatic carcinoma cells. CONCLUSIONS: The CD gene mediated by adenovirus with a high infectivity is efficient for gene therapy of pancreatic carcinoma cell lines. These data demonstrate the therapeutic efficacy of an enzyme prodrug strategy in experimental pancreatic cancer.
Assuntos
Adenoviridae/genética , Citosina Desaminase/genética , Vetores Genéticos/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/terapia , Linhagem Celular Tumoral , Clonagem de Organismos/métodos , Terapia Genética/métodos , Humanos , Pró-Fármacos/uso terapêutico , Resultado do TratamentoRESUMO
OBJECTIVE: To assess the diagnostic value of endoscopic pancreatic duct brushing in detecting mutation of the K-ras gene at codon 12 in cytologic specimens from patients with pancreatic cancer. METHODS: Thirty-five patients treated at Changhai Hospital, Shanghai between 1999 and 2001 were enrolled. Their cells obtained by pancreatic duct brushing during endoscopic retrograde cholangiopancreatography (ERCP) were suspended with phosphate buffer solution (PBS). DNA of the cells was extracted and mutation of the K-ras gene at codon 12 detected by means of PCR-SSCP. RESULTS: The K-ras gene mutation rate of pancreatic cancer was 70%, which was higher than that of chronic pancreatitis (14%, P<0.05). K-ras gene mutation was not found in patients with pancreatic cystocarcinoma and duodenum carcinoma. As to the location of pancreatic cancer, no significant difference was observed between the head, the body and tail. The sensitivity, specificity, accuracy of pancreatic duct brushing in detecting pancreatic cancer was 70%, 94%, and 83%, respectively. CONCLUSION: K-ras analysis of pancreatic brushing samples is helpful in the diagnosis of patients with early pancreatic cancer.
Assuntos
Colangiopancreatografia Retrógrada Endoscópica/métodos , Genes ras/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Mutação Puntual , Adulto , Idoso , Idoso de 80 Anos ou mais , Colangiopancreatografia Retrógrada Endoscópica/efeitos adversos , Doença Crônica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ductos Pancreáticos , Pancreatite/genética , Pancreatite/patologia , Polimorfismo Conformacional de Fita SimplesRESUMO
OBJECTIVE: The incidence of pancreatic cancer is increasing in China, and in many patients the surrounding lymphatics have already been invaded and there is blood-borne metastasis at the time of diagnosis. Additionally, pancreatic cancer is largely refractory to conventional therapies. Therefore, to improve its prognosis, it is important to resolve the problem of its growth. Angiotensin II type 1 receptor (AT1) stimulates the growth and angiogenesis of pancreatic cancer and a selective AT1 antagonist could inhibit these effects. The present study aimed to investigate the expression of AT1 in pancreatic cancer cell lines to provide the theoretical basis for its treatment. METHODS: The pancreatic cancer cell lines were SW1990, PaTu8988s and PANC-1. RT-PCR was used to detect the AT1 mRNA expression, and ABC immunocytochemical staining and SDS-PAGE were used to detect the expression of AT1 protein. RESULTS: Both AT1 mRNA and protein were expressed in all three cell lines. The AT1 protein was found on the cell membrane and in the cytoplasm of these cells. The AT1 protein (44 x 10(3)) was also demonstrated by SDS-PAGE. CONCLUSION: The results suggest that AT1 plays an important role in the growth of pancreatic cancer and its inhibition may be a therapeutic strategy.
Assuntos
Neoplasias Pancreáticas/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Linhagem Celular Tumoral , Humanos , RNA Mensageiro , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVE: To investigate the dynamic changes of mitogen-activated protein kinase (MAPK) signal transduction in rats with severe acute pancreatitis (SAP). METHODS: The SAP model was induced by infusing the bilio-pancreatic duct of 56 Sprague-Dawley rats with 5% sterile sodium taurocholate solution. The rats were randomly divided into seven groups: control group, 0.5 h postoperative group, 1 h group, 3 h group, 6 h group, 12 h group and 24 h group. Western blot analysis was used to determine the activities of p38 MAPK and c-Jun N-terminal kinase (JNK) in the pancreas and lungs. RESULTS: In the rats of the control group, basal p38MAPK activity could be detected but not that of JNK. After SAP was induced, the p38MAPK activity in the pancreas increased markedly and peaked at 3 h, but in the lung it peaked at 6 h. The p38MAPK activity in the pancreas and lungs was significantly higher than the basal activity at the 24 h time point. The activity of INK was only increased at the 12 h point and was not detectable at 24 h. CONCLUSION: The MAPK signal transduction pathway, in particular p38MAPK, plays an important role in the pathogenesis of SAP.
Assuntos
Pancreatite/fisiopatologia , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/farmacologia , Doença Aguda , Animais , Pulmão/enzimologia , Masculino , Pâncreas/enzimologia , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To investigate in rats the role of endothelin (ET)-1 gene expression in the development and progression of acute gastric mucosal lesions (AGML) induced by stress, and the effect of BQ-123 (a special ETA receptor antagonist) on the AGML. METHODS: A rat model of gastric ulcer induced by cold-restraint-stress (CRS) was used. ET-1 concentrations in the plasma and gastric mucosa were determined by radioimmunoassay (RIA), gastric mucosa blood flow (GMBF) was measured with a laser Doppler flow meter, the ulcer index (UI) was used to estimate the degree of gastric mucosa damage and the expression levels of ET-1 mRNA in the gastric mucosa were measured using dot blot and reverse transcription polymerase chain reaction (RT-PCR). Different doses of BQ-123 were administered via the left femoral vein prior to the stress in order to observe the effects of BQ-123 on the ET-1 concentrations in the plasma and gastric mucosa, the GMBF and the UI. RESULTS: Compared with the normal controls, the ET-1 concentrations in the plasma and gastric mucosa of the stressed rats were increased significantly (P < 0.05), the GMBF was decreased markedly (P < 0.01), and the UI increased dramatically (P < 0.01). There was a significant positive correlation between the gastric mucosal EF-1 concentration and the UI (r = 0.98, P < 0.01), and a significant negative correlation between the gastric mucosal ET-1 concentration and GMBF (r = -0.89, P < 0.01) and also between the UI and GMBF (r = -0.98, P < 0.01). The expression level of ET-1 mRNA in the gastric mucosa of the stressed rats increased significantly compared with that of the normal controls (P < 0.01), and there was a positive correlation between the expression of ET-1 mRNA and the ET-1 concentration in the gastric mucosa (r = 0.93, P < 0.01). Compared with the untreated animals, the GMBF was increased (P < 0.01) and the UI decreased significantly (P < 0.01) in the BQ-123-treated rats, and the dose of BQ-123 correlated with the degree of change in the GMBF and UI; however, the ET-1 concentrations of either the plasma or the gastric mucosa did not change markedly in the BQ-123-treated animals (P > 0.05). CONCLUSION: The present study showed that the level of expression of ET-1 mRNA and the synthesis of ET-1 in the gastric mucosa both increased significantly, which suggests that the increased concentration of endogenous ET-1 may be involved in the development and progression of stress ulcer (acute gastric mucosa lesion). The mechanism of action may be associated with a reduction of GMBF induced by ETAR-mediated vasoconstriction. BQ-123 can dose-dependently attenuate significantly the degree of damage to the gastric mucosa induced by stress, and may have therapeutic benefits for stress ulcer.
Assuntos
Endotelina-1/biossíntese , Mucosa Gástrica/metabolismo , Expressão Gênica , Úlcera Gástrica/metabolismo , Estresse Fisiológico/metabolismo , Doença Aguda , Animais , Temperatura Baixa , Antagonistas dos Receptores de Endotelina , Endotelina-1/genética , Mucosa Gástrica/irrigação sanguínea , Masculino , Peptídeos Cíclicos/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Restrição Física , Úlcera Gástrica/etiologia , Úlcera Gástrica/genética , Úlcera Gástrica/fisiopatologia , Estresse Fisiológico/genética , Estresse Fisiológico/fisiopatologiaRESUMO
OBJECTIVE: Visceral hypersensitivity is highly prevalent in most functional bowel disorders, such as irritable bowel syndrome (IBS), and activation of intestinal mast cells (MC) may play a role because they have been found in close proximity to gastrointestinal mucosal sensory nerve terminals containing neuropeptides and a bi-directional pathway connecting the central nervous system, gut, and MC has been demonstrated. The current study appraised the status of rectal visceral perception, as well as the changes in the MC and substance P (SP) in the intestinal mucosa of patients with IBS. METHODS: The study group comprised 42 patients with IBS and 19 healthy subjects who underwent anorectal manometry and rectal perception thresholds to balloon distension. The MC and the SP-ergic terminals in the mucosa were stained for respective histochemical and immunohistochemical investigations. The results were presented both qualitatively and quantitatively by color image analyzer, based on analysis of the intensity and area of stained fibrils. The structural relationship between the MC and nerve terminals was studied by electron microscopy, using an in situ embedding technique. RESULTS: The anorectal resting pressure, squeezing pressure and relaxation pressure were normal in both groups. The sensation threshold, defecation threshold and pain threshold in diarrhea-predominant IBS and the pain thresholds in constipation-predominant IBS were much lower than in the controls. Rectal compliance decreased in IBS. The number of MC in the terminal ileum, the ileocecal junction and the ascending colon was significantly elevated in IBS (P < 0.01), and the MC showed great variation. A significantly increased concentration of SP was found in the colon of the IBS patients compared with the controls. There was a positive correlation between the profiles of mucosal MC and the SP-ergic terminals, and MC were closely adjacent to SP-ergic terminals in the lamina propria. CONCLUSION: As altered rectal perception is present in almost all patients with IBS, it might be a reliable biological characteristic of the disease. Alterations in the MC and SP of the intestinal mucosa may be important factors in visceral hypersensitivity.
Assuntos
Síndrome do Intestino Irritável/fisiopatologia , Percepção , Reto/fisiologia , Adulto , Estudos de Casos e Controles , Defecação , Feminino , Humanos , Mucosa Intestinal/fisiologia , Masculino , Mastócitos/fisiologia , Pessoa de Meia-Idade , Limiar da Dor , Pressão , Reto/inervação , Substância P/análiseRESUMO
OBJECTIVE: To investigate the changes of the mast cells (MCs) and substance P (SP), and to elucidate their possible roles in visceral hypersensitivity in patients with irritable bowel syndrome (IBS). METHODS: In 22 diarrhea-predominant IBS, 20 constipation-predominant IBS and 19 controls, the biopsies were carried out from the terminal ileum, the ileocecal junction, the ascending colon, and the sigmoid colon. The MCs and the SP-ergic nerve terminals, SP receptor (SPR) cells were stained by histochemistry and immunohistochemistry respectively, and the results were investigated qualitatively and quantitatively by color image analyzer. The biopsies of the ICJ and the sigmoid colon were measured by radioimmunoassay. The structure relation between the MCs and SP-ergic terminals, SPR-ergic cells were studied through an ultramicroscopy using in situ embedding technique and a light microscopic study in serial sections respectively. RESULTS: The number of MCs in the terminal ileum, the ileocecal junction, and the ascending colon were significantly elevated in IBS patients (P < 0.01), and the MCs in IBS have great variations. Significantly increased the SP-ergic nerve terminals were found in patients with IBS of intestine compared with the control. The correlation between mucosal MC and the SP-ergic nerve terminals was found, and MCs were close to these terminals in lamina propria, which were demonstrated SP-ergic nerve terminals. Some MCs were demonstrated to be SPR-positive cells. CONCLUSIONS: The MCs and SP of intestinal mucosa may play a central role in the gut hypersensitivity in both motor response and visceral perception in IBS.