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1.
Cancer Invest ; 42(6): 527-537, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38965994

RESUMO

Despite the emergence of various treatment strategies for rectal cancer based on neoadjuvant chemoradiotherapy, there is currently a lack of reliable biomarkers to determine which patients will respond well to neoadjuvant chemoradiotherapy. Through collecting hematological and biochemical parameters data of patients prior to receiving neoadjuvant chemoradiotherapy, we evaluated the predictive value of systemic inflammatory indices for pathological response and prognosis in rectal cancer patients. We found that baseline GRIm-Score was an independent predictor for MPR in rectal cancer patients. However, no association was observed between several commonly systemic inflammation indices and long-term outcome.


Assuntos
Terapia Neoadjuvante , Neoplasias Retais , Humanos , Neoplasias Retais/patologia , Neoplasias Retais/terapia , Neoplasias Retais/imunologia , Masculino , Feminino , Pessoa de Meia-Idade , Terapia Neoadjuvante/métodos , Idoso , Quimioembolização Terapêutica/métodos , Prognóstico , Resultado do Tratamento , Adulto , Quimiorradioterapia/métodos
2.
Int J Colorectal Dis ; 35(6): 1155-1161, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32300884

RESUMO

PURPOSE: Investigation of the role of sedation during colonoscopy is meaningful as the advantages of colonoscopy performing with sedation are still controversial. METHODS: Medical records of patients who underwent colonoscopy in our institution were retrospectively analyzed. The sedation rate, adenoma detection rate (ADR), polyp detection rate (PDR), cecal intubation rate (CIR), iatrogenic colonic perforation rate (ICP) were calculated. RESULTS: A total of 48,838 colonoscopies (24,498 in males) dated from July 2007 to February 2017 were analyzed. The median age was 50 years (range 16-85 years). An overall sedation rate was 80.38%. The PDR was 26.77%, and was not statistically different between colonoscopy with or without sedation (26.67% vs 27.22, p = 0.474). ADR was 12.9% regardless of applying sedation or not (13.0% vs 12.44%, p = 0.337). The CIR was 87.42% in all examinations with an adjusted CIR of 90.34%, and was higher when performed with sedation than without sedation (88.92% vs 80.64%, p < 0.0001). Five cases (0.01%) of ICP were reported, all of which occurred in patients under sedation. CONCLUSIONS: The use of sedation is associated with increased CIR, but ADR and PDR remain unchanged with or without sedation. However, perforation rate, albeit very low, is significantly higher in sedated patients.


Assuntos
Adenoma/diagnóstico por imagem , Pólipos do Colo/diagnóstico por imagem , Colonoscopia/normas , Neoplasias Colorretais/diagnóstico por imagem , Sedação Consciente/estatística & dados numéricos , Sedação Profunda/estatística & dados numéricos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Ceco/diagnóstico por imagem , Colonoscopia/efeitos adversos , Colonoscopia/métodos , Sedação Consciente/efeitos adversos , Sedação Profunda/efeitos adversos , Detecção Precoce de Câncer , Feminino , Humanos , Perfuração Intestinal/etiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto Jovem
3.
Tumour Biol ; 37(3): 3441-9, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26449827

RESUMO

Rectal cancer, defined as a cancerous lesion of the colon distal to the rectosigmoid junction, is the fourth most common cancer cause of death globally. There were 474 patients with rectal cancer who underwent surgery between October 2007 and May 2013 enrolled in our center. Patients were respectively categorized by neoadjuvant therapy. This study aimed to explore the predictive factors that affected the Progression-free survival and overall survival of the patients with rectal cancer. Clinical characteristics of patients were compared with the groups and potential prognostic factors were analyzed by SPSS 19.0. In our study, neoadjuvant therapy increased the anus-retained rate (64.4 vs 53.4 % P = 0.016) and remission rate in the treatment group, compared to the non-treatment group (62.6 vs 34.8 %; P = 0.000). The neoadjuvant concurrent chemoradiotherapy, more operative duration, anus retained and micturition damaged are positive prognostic factors of PFS to patients. Poor differentiation, the tumor of ulcer, invasive, and pT4 stage, contributed the poor factors for PFS of patients (P < 0.05). Additionally, the patients with neoadjuvant concurrent chemoradiotherapy and adjuvant chemotherapy underwent the better prognosis of OS. Adjuvant chemotherapy cannot increase PFS of the patients who accepted neoadjuvant therapy after surgery get pCR, but can improve OS. The anus-retained and neoadjuvant radiotherapy, duration of surgery in rectal cancer have the positive correlation. Micturition damaged and neoadjuvant radiotherapy were positively correlated as well. In conclusion, adjuvant chemotherapy does not improve the PFS of patients with pCR to neoadjuvant therapy, but is good for OS. Further prospective and large population-based clinical studies are needed to establish clinical guidelines for the use of neoadjuvant therapy and adjuvant chemotherapy in patients with rectal cancer.


Assuntos
Neoplasias Retais/terapia , Idoso , Quimiorradioterapia , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante , Estadiamento de Neoplasias , Prognóstico , Neoplasias Retais/mortalidade , Neoplasias Retais/patologia , Estudos Retrospectivos
4.
Zhongguo Zhong Yao Za Zhi ; 40(9): 1825-30, 2015 May.
Artigo em Zh | MEDLINE | ID: mdl-26323157

RESUMO

A quantitative analytical method of ultra-high performance liquid chromatography (UPLC) was developed for simultaneously determining twelve components in Tibetan medicine Zuozhu Daxi. SIMPCA 12.0 software was used a principal component analysis PCA) and partial small squares analysis (PLSD-DA) on the twelve components in 10 batches from four pharmaceutical factories. Acquity UPLC BEH C15 column (2.1 mm x 100 mm, 1.7 µm) was adopted at the column temperature of 35 °C and eluted with acetonitrile (A) -0.05% phosphate acid solution (B) as the mobile phase with a flow rate of 0. 3 mL · min(-1). The injection volume was 1 µL. The detection wavelengths were set at 210 nm for alantolactone, isoalantolactone and oleanolic; 260 nm for trychnine and brucine; 288 nm for protopine; 306 nm for protopine, resveratrol and piperine; 370 nm for quercetin and isorhamnetin. The results showed a good separation among index components, with a good linearity relationship (R2 = 0.999 6) within the selected concentration range. The average sample recovery rates ranged between 99.44%-101.8%, with RSD between 0.37%-1.7%, indicating the method is rapid and accurate with a good repeatability and stability. The PCA and PLSD-DA analysis on the sample determination results revealed a great difference among samples from different pharmaceutical factories. The twelve components included in this study contributed significantly to the quantitative determination of intrinsic quality of Zuozhu Daxi. The UPLC established for to the quantitative determination of the twelve components can provide scientific basis for the comprehensive quality evaluation of Zuozhu Daxi.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Plantas Medicinais/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Controle de Qualidade , Tibet
5.
Front Oncol ; 13: 1201544, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37456230

RESUMO

Objectives: Locally advanced rectal cancer (LARC) has a high risk of distant metastasis (DM). Currently, many treatment courses of LARC have arisen, but patients' DM status has not significantly improved. This study was designed to compare the effect between preoperative regional transarterial chemoembolization combined with neoadjuvant chemoradiotherapy and standard neoadjuvant therapy on preventing DM in patients with LARC. Methods: A total of 81 LARC patients between July 2013 and May 2018 were enrolled in this retrospective study. Among them, 44 patients received preoperative regional transarterial chemoembolization combined with concurrent chemoradiotherapy (the interventional group), and 37 patients received only neoadjuvant chemoradiotherapy (the control group). The baseline data; preoperative toxicities; postoperative DM rate within 1, 2, and 3 years; and postoperative complications were compared between the two groups. Results: All patients successfully completed their treatments. There were no significant differences between the two groups in age, gender, tumor size, distance between the tumor and anal verge, CEA level, lymphovascular invasion, or tumor stage before treatment. The pathological T staging post-treatment in the interventional group was significantly reduced compared to that of the control group (p = 0.025). There were no significant differences between groups in DM rates within 1 and 2 years after surgery. In terms of DM rate within 3 years after surgery, the interventional group was significantly lower than that of the control group (9.1% vs. 29.7%, p = 0.036). Conclusion: Preoperative regional transarterial chemoembolization combined with concurrent chemoradiotherapy may play an important role in reducing postoperative DM in LARC.

6.
Quant Imaging Med Surg ; 11(7): 3120-3132, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34249639

RESUMO

BACKGROUND: Acute arterial occlusive mesenteric ischemia with transmural intestinal necrosis (TIN) is a fatal disease, which is difficult to diagnose on multidetector computed tomography (MDCT). The aim of the present study was to determine the relationship of superior mesenteric artery (SMA) thrombus density with TIN on MDCT in patients with acute mesenteric ischemia (AMI) due to SMA thromboembolism. METHODS: In this retrospective study, 33 patients who underwent abdominal MDCT and angiography for AMI due to SMA thromboembolism were divided into two groups: the AMI with TIN group and the AMI without TIN group. We analyzed the relationships of clinical characteristics, qualitative MDCT signs, and SMA thrombus density with TIN. The SMA thrombus density was measured on non-contrast MDCT. Univariate and multivariate analyses were performed to determine the risk factors for predicting TIN. The diagnostic performances of risk factors were evaluated by receiver-operating characteristic (ROC) curve analysis. RESULTS: Of the patients with AMI enrolled in this study, 33.3% (11/33) were diagnosed with TIN. Peritonitis (P=0.042), bowel wall thinning (P=0.033), and pneumatosis/portomesenteric gas (P=0.010) were significantly associated with TIN. AMI patients with TIN exhibited a higher SMA thrombus density than AMI patients without TIN [41.2±6.1 vs. 34.2±3.0 Hounsfield unit (HU), P=0.003]. Multivariate analysis showed that SMA thrombus density was an independent predictor of TIN [P=0.044, hazard ratio (HR): 1.82, 95% confidence interval (CI): 1.02-3.25]. For diagnosing AMI with TIN, the area under the ROC curve (AUC) of SMA thrombus density (0.83) was larger than those of peritonitis (0.68), bowel wall thinning (0.66), and pneumatosis/portomesenteric gas (0.71). CONCLUSIONS: In patients with AMI, erythrocyte-rich thrombus blocking the SMA trunk which has a higher density on MDCT is prone to the occurrence of TIN compared with erythrocyte-scarce thrombus with a lower density. SMA thrombus density could be an independent risk factor for TIN in patients with AMI due to SMA thromboembolism.

7.
J Zhejiang Univ Sci B ; 21(8): 657-667, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32748581

RESUMO

Camellia oil has become an important plant oil in China in recent years, but its effects on non-alcoholic fatty liver disease (NAFLD) have not been documented. In this study, the effects of camellia oil, soybean oil, and olive oil on NAFLD were evaluated by analyzing the fatty acid profiles of the plant oils, the serum lipids and lipoproteins of rats fed different oils, and by cytological and ultrastructural observation of the rats' hepatocytes. Analysis of fatty acid profiles showed that the polyunsaturated fatty acid (PUFA) n-6/n-3 ratio was 33.33 in camellia oil, 12.50 in olive oil, and 7.69 in soybean oil. Analyses of serum lipids and lipoproteins of rats showed that the levels of total cholesterol and low-density lipoprotein cholesterol in a camellia oil-fed group (COFG) were lower than those in an olive oil-fed group (OOFG) and higher than those in a soybean oil-fed group (SOFG). However, only the difference in total cholesterol between the COFG and SOFG was statistically significant. Cytological observation showed that the degree of lipid droplet (LD) accumulation in the hepatocytes in the COFG was lower than that in the OOFG, but higher than that in the SOFG. Ultrastructural analysis revealed that the size and number of the LDs in the hepatocytes of rats fed each of the three types of oil were related to the degree of damage to organelles, including the positions of nuclei and the integrity of mitochondria and endoplasmic reticulum. The results revealed that the effect of camellia oil on NAFLD in rats was greater than that of soybean oil, but less than that of olive oil. Although the overall trend was that among the three oil diets, those with a lower n-6/n-3 ratio were associated with a lower risk of NAFLD, and the effect of camellia oil on NAFLD was not entirely related to the n-6/n-3 ratio and may have involved other factors. This provides new insights into the effect of oil diets on NAFLD.


Assuntos
Camellia , Hepatopatia Gordurosa não Alcoólica/etiologia , Óleos de Plantas/administração & dosagem , Animais , Camellia/química , Ácidos Graxos/análise , Hepatócitos/patologia , Hepatócitos/ultraestrutura , Gotículas Lipídicas/fisiologia , Lipídeos/sangue , Masculino , Hepatopatia Gordurosa não Alcoólica/patologia , Ratos , Ratos Sprague-Dawley
8.
Br J Radiol ; 93(1116): 20190605, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32886528

RESUMO

OBJECTIVES: To identify risk factors of geriatrics index of comorbidity (GIC) and multidetector CT (MDCT) findings for predicting mortality in patients with acute mesenteric ischemia (AMI) due to superior mesenteric artery (SMA) thromboembolism. METHODS: 33 patients with AMI due to SMA thromboembolism underwent abdominal MDCT and angiography. Patients' comorbidities and MDCT findings of ischemic bowel/mesenteric injuries, regions of SMA involved by thromboembolism, and degrees of SMA stenosis were retrospectively reviewed. The comorbidities were classified into 1-4 levels according to GIC. The association of MDCT signs and GIC classification with mortality were analyzed. Diagnostic performances of risk factors associated with mortality were evaluated by receiver operating characteristic (ROC) curve analyses. RESULTS: Eighteen patients (54.5%) died during hospitalization or follow-up, including one patient with class 1, two patients with class 2, eight patients with class 3, and seven patients with class 4 according to GIC. Three risk factors significantly associated with mortality were identified, including pneumatosis and/or portomesenteric venous gas (PPMVG) (p = 0.017), four regions of SMA involved by thromboembolism (region I + II + III + IV) (p = 0.036), and class 3 + 4 of comorbidities (p = 0.001). The sensitivity and specificity of PPMVG, region I + II + III + IV, class 3 + 4 of comorbidities, and the three risk factors combined for diagnosing mortality were 33.3 and 100%, 27.8 and 100%, 83.3 and 73.3%, and 88.9 and 73.3%, respectively. The areas under the ROC curve (AUC) of the three risk factors combined (0.88) and class 3 + 4 of comorbidities (0.78) were larger than that of PPMVG (0.67) and region I + II + III + IV (0.64). The mortality rate rose from 15.4% in patients without risk factor to 66.7%, 100%, and 100% in patients with one, two, and three factors, respectively. CONCLUSION: Three risk factors for mortality were identified in patients with AMI due to SMA thromboembolism, including PPMVG and four regions of SMA involved by thromboembolism on MDCT images, and class 3 + 4 of comorbidities. Close monitoring of these risk factors could possibly lower the mortality. ADVANCES IN KNOWLEDGE: Risk factors based on GIC and MDCT findings may be used to predict mortality in patients with AMI. Close monitoring of these risk factors could possibly lower the mortality.


Assuntos
Isquemia Mesentérica/diagnóstico por imagem , Isquemia Mesentérica/mortalidade , Tomografia Computadorizada Multidetectores , Idoso , Idoso de 80 Anos ou mais , Feminino , Avaliação Geriátrica , Humanos , Masculino , Isquemia Mesentérica/complicações , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Retrospectivos , Fatores de Risco
9.
World J Clin Cases ; 8(23): 5976-5987, 2020 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-33344596

RESUMO

BACKGROUND: Abdominal ventral rectopexy (AVR) with colectomy is controversial in the treatment of obstructed defecation syndrome (ODS). Literature data on this technique for ODS are very limited. AIM: To evaluate the safety and efficacy of AVR with colectomy for selected patients with ODS. METHODS: Consecutive patients who underwent AVR with colectomy for ODS were identified prospectively from 2016 to 2017 in our department. Patient demographics, perioperative surgical results, and postoperative follow-up outcomes were collected and analyzed. Long-term follow-up was evaluated with standardized questionnaires. The severity of symptoms was assessed by the objective Wexner Constipation Score (WCS) and ODS Score. The quality of life was assessed by the Patients Assessment of Constipation Quality of Life score. Functional outcome was compared pre- and post-operatively for each patient. The primary outcomes were determined by the improvement in symptoms and quality of life. Secondary outcome measures were operating time, postoperative length of stay, morbidity and mortality, improvement of pelvic floor structure, and patient satisfaction. RESULTS: Four patients underwent robotic-assisted surgery, and two patients underwent a laparoscopic-assisted procedure. The mean operating time for the robotic approach was 243 min (range 160-300 min), and the mean operating time for the laparoscopic approach was 230 min (range 220-240 min). The mean postoperative length of stay was 8.2 d (range 6-12 d). There was no conversion to open procedure and no postoperative mortality. No urinary retention, wound infection, prolonged ileus, pelvic infection and anastomosis leakage occurred. Six patients were followed up for 36 mo. The WCS, ODS, and Patients Assessment of Constipation Quality of Life score improved significantly postoperatively (P < 0.05). The WCS and ODS scores showed the best remission and stabilization at 6 to 12 mo after surgery. There was no recurrence or novel constipation after surgery. None of the patients used laxative medication. CONCLUSION: Robotic and laparoscopic-assisted ventral rectopexy with colectomy is a safe and effective procedure for selected patients with ODS. However, comprehensive preoperative evaluation and careful patient selection are essential.

10.
J Zhejiang Univ Sci B ; 19(12): 960-972, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30507079

RESUMO

Royal jelly (RJ) from honeybee has been widely used as a health promotion supplement. The major royal jelly proteins (MRJPs) have been identified as the functional component of RJ. However, the question of whether MRJPs have anti-senescence activity for human cells remains. Human embryonic lung fibroblast (HFL-I) cells were cultured in media containing no MRJPs (A), MRJPs at 0.1 mg/ml (B), 0.2 mg/ml (C), or 0.3 mg/ml (D), or bovine serum albumin (BSA) at 0.2 mg/ml (E). The mean population doubling levels of cells in media B, C, D, and E were increased by 12.4%, 31.2%, 24.0%, and 10.4%, respectively, compared with that in medium A. The cells in medium C also exhibited the highest relative proliferation activity, the lowest senescence, and the longest telomeres. Moreover, MRJPs up-regulated the expression of superoxide dismutase-1 (SOD1) and down-regulated the expression of mammalian target of rapamycin (MTOR), catenin beta like-1 (CTNNB1), and tumor protein p53 (TP53). Raman spectra analysis showed that there were two unique bands related to DNA synthesis materials, amide carbonyl group vibrations and aromatic hydrogens. These results suggest that MRJPs possess anti-senescence activity for the HFL-I cell line, and provide new knowledge illustrating the molecular mechanism of MRJPs as anti-senescence factors.


Assuntos
Senescência Celular/efeitos dos fármacos , Ácidos Graxos/química , Fibroblastos/citologia , Pulmão/citologia , Animais , Abelhas , Bovinos , Linhagem Celular , Proliferação de Células , Meios de Cultura , Relação Dose-Resposta a Droga , Fibroblastos/efeitos dos fármacos , Humanos , Proteínas de Insetos/química , Pulmão/efeitos dos fármacos , Albumina Sérica/metabolismo , Análise Espectral Raman , Superóxido Dismutase-1/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Proteína Supressora de Tumor p53/metabolismo , beta Catenina/metabolismo
11.
Pest Manag Sci ; 74(7): 1720-1727, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29363888

RESUMO

BACKGROUND: The use of thelytokous Wolbachia-infected Trichogramma (parasitic wasps) has long been considered as a way to enhance the efficacy of biocontrol. However, Wolbachia can affect the host physiology. We compared decision-making between bisexual and thelytokous Wolbachia-infected lines of Trichogramma dendrolimi Matsumura regarding behavior toward fresh and old eggs of Corcyra cephalonica at 25 ± 1 °C and 70 ± 5% relative humidity. RESULTS: The behavioral patterns and sequences of the two lines were basically the same. The durations of various behavioral patterns and values of fitness indicators of the bisexual line on fresh eggs were generally significantly shorter and better, respectively, than on old eggs, whereas the thelytokous line behaved similarly toward the two types of eggs, and differences in most fitness indicators between fresh and old eggs were not significant. On fresh eggs, the durations of various behaviors in the bisexual line were generally significantly shorter than in the thelytokous line and the fitness indicators were generally significantly better. CONCLUSION: Wolbachia affected the fitness of T. dendrolimi negatively. The potential of the thelytokous line as a biocontrol agent would not be as good as that of the bisexual line when decision-making only is considered. Therefore, further evaluations need to be carried out before the thelytokous line can be used in practical biocontrol. © 2018 Society of Chemical Industry.


Assuntos
Mariposas/parasitologia , Controle Biológico de Vetores , Vespas/microbiologia , Vespas/fisiologia , Wolbachia/fisiologia , Animais , Tomada de Decisões , Feminino , Controle de Insetos
13.
Mayo Clin Proc ; 91(4): 456-68, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27046523

RESUMO

OBJECTIVE: To evaluate the efficacy and safety of lubiprostone in the treatment of chronic idiopathic constipation (CIC) and irritable bowel syndrome with constipation (IBS-C). PATIENTS AND METHODS: We performed a literature search of the MEDLINE, Cochrane, Google Scholar, and ClinicalTrials.gov databases (from January 1, 2005, through January 31, 2015). Relevant studies meeting the inclusion criteria were manually searched by 2 independent reviewers. Efficacy outcomes evaluated at 1 week, 1 month, and 3 months of intervention were weekly frequency of spontaneous bowel movements, severity of constipation, consistency of stools, degree of abdominal pain/discomfort, degree of straining, and abdominal bloating. RESULTS: Of 246 studies identified, data from 9 trials comprising 1468 patients (63.6%) in the lubiprostone group and 841 (36.4%) in the placebo group were analyzed. We found that lubiprostone treatment significantly improved the severity of constipation, stool consistency, abdominal pain, degree of straining, and abdominal bloating at 1 week (P≤.03) and 1 month (P≤.004), except for abdominal pain at 1 month, which was similar to that when treated with placebo (P=.21). At 3 months, except for abdominal bloating (P=.03), there was no difference between lubiprostone and placebo groups in all other outcomes (P≥.05). Adverse effects such as nausea, vomiting, and diarrhea were common (incidence rate, 2.4%-75%); however, the incidence of serious adverse effects was low (<5%) and was mostly unrelated to lubiprostone treatment. CONCLUSION: Lubiprostone is a safe and efficacious drug for the treatment of chronic idiopathic constipation and irritable bowel syndrome with constipation, with limited adverse effects in 3 months of follow-up.


Assuntos
Dor Abdominal/tratamento farmacológico , Doença Crônica/tratamento farmacológico , Constipação Intestinal/tratamento farmacológico , Fármacos Gastrointestinais/uso terapêutico , Síndrome do Intestino Irritável/tratamento farmacológico , Laxantes/uso terapêutico , Lubiprostona/uso terapêutico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ensaios Clínicos Controlados Aleatórios como Assunto , Adulto Jovem
14.
World J Gastroenterol ; 11(18): 2838-40, 2005 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-15884135

RESUMO

AIM: To study the method of dissociation, culture and investigate its morphologic changes in vitro of interstitial cells of Cajal (ICC). METHODS: Enzymatic digestion and Ficoll density centrifugation were used to dissociate ICC from the ileal segment of mice. Factors including contamination, Ca(2+), Mg(2+) and collagenase, and stem cell factor, etc., were investigated. ACK2, the antibody of c-kit, was used to identify the cultured ICC. Both light microscope and fluorescence microscope were used to observe the changes of ICC in vitro. RESULTS: The method for dissociation and culture of ICC in vitro was successfully established. After 24 h, cultured ICC exhibited a few axis-cylinders, and longer axis-cylinders were observed to form synapse of each other after 3 d. More widespread connections formed within 7 d in vitro. The changes of its morphologic character were obvious within 7 d; however, there were no obvious morphologic changes after 30 d. CONCLUSION: Many factors can influence the dissociation and culture of ICC.


Assuntos
Relógios Biológicos , Técnicas de Cultura de Células , Separação Celular , Trato Gastrointestinal/citologia , Trato Gastrointestinal/fisiologia , Músculo Liso/citologia , Músculo Liso/fisiologia , Animais , Células Cultivadas , Camundongos , Camundongos Endogâmicos BALB C , Fatores de Tempo
16.
Zhonghua Wei Chang Wai Ke Za Zhi ; 10(2): 119-23, 2007 Mar.
Artigo em Zh | MEDLINE | ID: mdl-17380448

RESUMO

OBJECTIVE: To investigate the construction of the green fluorescent protein (GFP) labeled recombinant adenovirus containing human stem cell leukemia (hSCL) and its distribution and efficiency in mice with interstitial cells of Cajal (ICC) loss. METHODS: The recombinant adenovirus Ad-GFP/SCL was constructed by Ad-Easy system based on the homologous recombination in bacteria, then 1.6 x 10(9) PFU of recombinant adenoviruses were injected into Balb/c mice with ICC loss via the tail vein. In vivo distribution and efficiency of recombinant adenoviruses mediated hSCL were observed by GFP under the fluorescent microscope at different phases. The expression of SCL gene was measured by RT-PCR method. The damages were observed in different organs by HE staining. RESULTS: The recombinant adenovirus containing hSCL was quickly constructed by homologous recombination in bacteria using Ad-Easy system. Under the fluorescent microscope, GFP was revealed in heart, lung, liver, spleen, kidney, small intestine and large intestine of mice with ICC loss at different phases. No obvious damages were observed in various visceral organs by HE staining. RT-PCR showed SCL mRNA expression in various visceral organs at different levels. CONCLUSIONS: Construction of adenovirus vector by the homologous recombination in bacteria is an efficient and time saving method, and a high titer of adenovirus is able to mediate the safe and stable expression of SCL gene in mice with ICC loss. This finding will make further gene therapy in mice with STC possible.


Assuntos
Adenoviridae/genética , Terapia Genética , Proteínas de Fluorescência Verde/genética , Leucemia Mieloide Aguda/genética , Adenoviridae/metabolismo , Animais , Constipação Intestinal/terapia , Feminino , Vetores Genéticos , Proteínas de Fluorescência Verde/metabolismo , Humanos , Células Intersticiais de Cajal/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Recombinação Genética , Transdução Genética
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