Activation of Sonic hedgehog signaling in ventricular cardiomyocytes exerts DNA protection against hypoxia / 中国药理学通报

Aim To investigate the protective roles of sonic hedgehog( Shh) signaling pathway in hypoxia-in-duced DNA damage with the neonatal rat cardiomyo-cytes. Methods The hypoxia model on neonatal car-diomyocytes was established with one to two days old Sprague Dawley rats by deprivation of oxygen and glu-cose ( OGD) . After pretreated with Shh pathway ago-nist SAG1.3 or antagonist GANT61, the survival rates of cardiomyocytes were assayed by MTT after OGD 6 hours or 12 hours. The protein levels of Shh pathway, phosphorylated histone H2AX at serine 139 (γH2AX), phosphorylated ATM (p-ATM), phospho-rylated p53 ( p-p53 ) , cleaved-caspase-3, Bcl-2 and Bax were detected by Western blot. The γH2AX foci was detected by immunofluorescence. Results Com-pared to control group, the protein expression of γH2AX, p-ATM, cleaved-caspase-3, p-p53 in OGD cardiomyocytes significantly increased, and Bcl-2/Bax ratio proportionally decreased. Particularly, the ex-pression of γH2AX, p-ATM was highest at OGD 6 h, and then gradually declined after OGD 12 h. After SAG1.3 pretreatment, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 dramatically de-creased and the Bcl2/Bax ratio increased in OGD 6 h or OGD 12 h cardiomyocytes. On the contrary, in GANT61 pretreatment group, the expression of γH2AX, p-ATM, cleaved-caspase-3 and p-p53 signifi-cantly increased and the Bcl-2/Bax ratio decreased compared to the OGD 6 h or OGD 12 h cardiomyo-cytes. Conclusion The activation of Shh pathway protects cardiomyocytes against hypoxia-induced apop-tosis through inhibition of DNA damage.

Crosstalk of Hedgehog/Gli and PI3k/Akt Pathway Promotes Epithelial-mesenchymal Transition in Gastric Adenocarcinoma / 中国医科大学学报

Objective To investigate the tumor molecular mechanism of Hedgehog/Gli in promoting the epithelial-mesenchymal transition (EMT) in gastric cancer AZ521 cells. Methods After 24 h of treatment with GANT61,the mRNA expression of Gli1,Gli2, N-cadherin,and E-cadherin in the AZ521 cell line were detected by real-time fluorescence quantitative PCR. A Western blotting assay was conducted to determine the expression of the above cytokines,p-AKT and AKT. The effect of GANT61 on invasion was observed by transwell assay. N-Shh stimulation of the Hedgehog pathway was conducted to confirm the changes in these cytokines. Results GANT61 significantly downregulated the mRNA expression of Gli1,Gli2,and N-cadherin,but upregulated E-cadherin mRNA expression. The Western blotting assay revealed that GANT61 downregulated the protein expression of Gli1,Gli2,p-AKT,and N-cadherin,but upregulated E-cadherin expression. Furthermore,GANT61 inhibited the invasion. N-Shh proteins up-regulated Gli1,Gli2,and N-cadherin mRNA,protein expression and p-AKT protein expression,but downregulated E-cadherin mRNA and protein expressions. N-Shh promoted the invasion of tumor cells. Conclusion Downregulation of Gli1 and Gli2 can inhibit the invasion and metastasis in gastric cancer cells,which may be related to the promotion of EMT by Gli through the PI3K/AKT pathway.

Study on role of GANT-61 as Gli inhibitor on epithelial-mesenchymal transition of lung cancer / 重庆医学

Objective To research the effect of GANT61 on epithelial‐mesenchymal transition (EMT) in human lung cancer H1703 and A549 cells lines ,and to preliminarily investigate its action mechanism .Methods DMSO was used as the control(DMSO group) .After treating H1703 and A549 cells with GANT61 for 24 h ,the gene changes of Gli‐1 ,Gli‐2 ,E‐cadherin and Vimentin were detected by using the real time fluorescence quantitative PCR method .The influence of GANT61 on the expression of E‐cad‐herin and Vimentin protein after acting on H1703 and A549 cells was observed by using the Western blotting assay .The scratch healing test was performed to evaluate the effect of GANT 61 on the tumor cell invasion ability after acting on H1703 and A549 cells .Results The real time fluorescence quantitative PCR showed that ,compared with the DMSO group ,GANT61 down‐regulated the mRNA expression of Gli‐1 ,Gli‐2 and Vimentin mRNA of H1703 and A549 cell lines and elevated the expression of E‐cadherin protein(P<0 .01);the Western blotting showed that GANT61 down‐regulated the expression of Vimentin of H1703 and A549 cell lines ,and elevated the expression of E‐cadherin(P<0 .01);the scratch healing test revealed the invasion ability of H 1703 and A549 cells in the GANT61 treatment group was significantly decreased (P<0 .01) .Conclusion EMT in lung cancer is related with aber‐rant activations of Gli1 and Gli2 in Hedgehog signal transduction pathway ,GANT61 could influence the EMT ability in lung cancer cells by down‐regulating the expression of Gli‐1 and Gli‐2 .Gli could become a new molecular target for inhibiting the lung cancer cell metastasis.

GANT61 as an inhibitor of Gli inhibits growth and invasion of esophageal adenocarcinoma / 天津医药

Objective To study the inhibitory effects of GANT61, as an inhibitor of Gli, on the growth of human esophageal adenocarcinoma cell lines OE19 and OE33, and their mechanisms thereof. Methods After treating with different concentrations of GANT61(30,20,13.333 3,8.888 8,5.925 9,3.950 6,2.633 7,1.755 8,1.170 5μmol/L),the cell viabilities of OE19 and OE33 were detected by MTS method, which expressed by IC50. The Gli1and Gli2 mRNA expressions treated with GANT61(10 μmol/L GANT61) or DMSO for 24 h were detected in OE19 and OE33 cell lines by real time fluorescence quantitative PCR. The protein expressions of Gli1, Gli2 and CyclinD1 treated with GANT61 or DMSO for 24 h were detected in OE19 and OE33 cell lines by Western blot assay. Transwell invasion assay was performed to evaluate the inhibiting effect on OE19 and OE33 cell invasion by the treatment of GANT61 or DMSO. Results The IC50 of GANT61 was 8.08μmol/L in OE19 and 9.65μmol/L in OE33 cells. Compared with DMSO group, Gli1 and Gli2 mRNA expressions and Gli1,Gli2 and CyclinD1 protein expressions were significantly decreased in OE19 and OE33 cells of GANT61 group (P<0.05). The number of penetrating cells was significantly reduced in OE19 and OE33 cells of GANT61 group compared with that of DMSO group (P<0.01). Conclusion GANT61 can inhibit the growth and invasion of esophageal neoplasms cells by down-regulating Gli1 and Gli2 mRNA expression,which indicates that Hedgehog signaling pathway may play an important role in carcinogenesis and progression of esophageal adenocarcinoma.

Laparoscopic sleeve excision of lesser curvature gastrointestinal autonomic nerve tumour.

Gastrointestinal autonomic nerve tumors (GANT) are uncommon stromal tumors of the intestinal tract. Their histological appearance is similar to that of other gastrointestinal stromal tumors. We report probably the first laparoscopic excision of this rare tumor from our geographical region and compare our findings with the available case reports in the medical literature. A 38 year old male patient undergoing a routine health check was diagnosed with an exophytic growth on the lesser curvature of stomach on computerized tomography and underwent an initial endoscopic biopsy which was inconclusive. He underwent a laparoscopic sleeve excision of the GANT on the lesser curvature. Radical surgical resection of gastrointestinal autonomic nerve tumors seems to be the only available curative approach and survival seems possible even in large tumors.

Clinical analysis of 16 cases with complete rectal prolapse undergoing Gant-Miwa and anal encircling procedure with Xiaozhiling injection / 国际外科学杂志

Objective To evaluate the efficacy of Gant-Miwa and anal encircling procedure with Xiaozhiling injection for complete rectal prolapse.Methods Clinical data of 16 patients with complete rectal prolapse undergoing Gant-Miwa and anal encircling procedure with Xiaozhiling injection between Feb.2011 and Nov.2014 were retrospectively analyzed.Results All patients were operated successufully and theirs prolapse symptons disappeared.Urinary retention was found in 3 cases,2 cases were observed a replase who were infected in the area of the anal operation,1 cases was found delayed healing of perianal incision,and 2 cases were happened constipation duing to the rectal stenosis.Complications wasn't happened such as bleeding,perianal abscess,anal fistula,and so on.Thirteen cases were followed-up of average 2.2 years and 2 cases were observed postoperative recurrance.Conclusions Gant-Miwa and anal encircling procedure with Xiaozhiling injection for the treatment of complete rectal prolapse is a procedure with minimally trauma,fewer complications and low recurrence rate,which is worth promoting after innovation and improvement.

Schwannoma de recto. Reporte de un caso / Schwannoma of rectum. Case report

Se presenta el caso clínico de un paciente de 30 años de edad que consulta por un cuadro de pujos y tenesmo rectal de dos meses de evolución. Al examen físico se constata una lesión submucosa en recto inferior de 3 cm de diámetro aproximado. Se realiza RMI de alta resolución con difusión sin imágenes de restricción por lo cual se decide tratamiento quirúrgico mediante un abordaje transanal a través del plano interesfintérico. El análisis Inmunohistoquímico de la pieza quirúrgica muestra positividad difusa para Proteína S-100 y Vimentina y negativa para otros marcadores, hallazgos compatibles con Schwannoma rectal.

The clinical case of a 30 year old who consults for pushing and rectal tenesmus of over two months of evolution is presented. On physical examination a small submucosal lesion in the lower rectum is found. RMI is performed with diffusion which show no images restraining which surgical treatment is decided by a transanal approach. Immunohistochemical analysis of the surgical specimen shows diffuse positivity for S-100 protein and vimentin, findings consistent with rectal schwannoma.

Tumores mesenquimales diagnosticados en la unidad de gastroenterología del HMPC entre los años 2007 aL 2009 / Mesenchymal tumors diagnosed in the gastroenterology unit of HPMC from 2007 to 2009

Se evaluaron las historias médicas con diagnóstico de tumores mesenquimales confirmados por análisis inmunohistoquímico entre los años 2007 y 2009. El objetivo fue describir a la población según características clínicas, epidemiológicas, diagnósticos por imágenes y análisis anatomopatológico. Se obtuvieron 11 pacientes con diagnósticos de GIST(7), Leiomiomas(2), GANT(1) y Leiomiosarcoma(1). Del género femenino (82%) y masculino (18%). Con un promedio de 55 años de edad. Clínicamente presentaron dolor abdominal (45%), mareos (27%), pirosis (9%) y disfagia (1%). El 18% permaneció asintomático. Entre los signos encontrados figuran: melena (36%), pérdida de peso (27%), palidez cutánea (9%) y vómitos (9%). El 36% de los pacientes no presentaron hallazgos al examen físico. Se realizó ultrasonido abdominal en el 100% de los pacientes, con hallazgos patológicos relacionados con el tumor en el 27 % de los casos. Al 90% de los pacientes se les realizó una Endoscopia Digestiva Superior, el 82% de los hallazgos se describieron como Tumores Submucosos y 9% se reportó como normal. Las lesiones se ubicaron en el estómago (60%), 27% en el intestino delgado ( duodeno y yeyuno); 9% en esófago y 9% en retroperitoneo. El ultrasonido endoscópico fue practicado al 82% de los pacientes, con un porcentaje de aciertos diagnósticos en el 90% de los tumores localizados en esófago, estómago e intestino delgado, y de 87,5% si se incluye el tumor retroperitoneal. La Tomografía Computada (TC) de abdomen fue practicada en 45% de los pacientes, con hallazgos patológicos en el 100% de los casos, descritos como Lesiones Ocupantes de Espacio (LOE), en estómago, intestino, esófago y retroperitoneo; y en el 20% de los pacientes se encontró enfermedad metastásica hepática Recibieron tratamiento quirúrgico el 82%, el 18% restante no se realizó por contraindicación médica o se encuentran en espera del procedimiento. Y un 18% de los pacientes recibieron tratamiento médico con Imatinib...

We evaluated the clinical records with a diagnosis of mesenchymal tumors confirmed by immunohistochemical analysis, from 2007 to 2009. The objective was to describe the population according to clinical and epidemiological features, diagnostic imaging and histopathological analysis. We found 11 patients with diagnoses of GIST(7), leiomyoma(2), leiomyosarcoma(1) and GANT(1); (82%) female and (18%) male, with a mean age of 55 years. They clinically presented abdominal pain (45%), dizziness (27%), heartburn (9%) and dysphagia (1%). 18% remained asymptomatic. Among the symptoms were: melena (36%), weight loss (27%), paleness (9%) and vomiting (9%). 36% of the patients had no findings at physical examination. Abdominal ultrasound was performed in 100% of the patients, with pathological findings related to the tumor in 27% of cases. In 90% of patients an upper digestive endoscopy was performed. 82% of findings were described as submucosal tumors and 9% was reported as normal. Lesions were located in stomach (60%); 27% in small bowel (duodenum and jejunum), 9% in esophagus and 9% in retroperitoneum. Endoscopic ultrasound was performed to 82% of patients, with a diagnostic accuracy of 90% for the tumors located in esophagus, stomach and small bowel; and 87,5% if the retroperitoneal tumor is included. The abdomen Computed Tomography (CT) was performed in 45% of patients with pathological findings in 100% of the cases, described as Space occupying lesion (SOL) in stomach, intestine, esophagus and retroperitoneum; and, in 20% of patients metastatic liver cancer was found. 82% received surgical treatment, the remaining 18% was not performed due to contraindication or are waiting for the procedure. And 18% of patients received medical therapy with Imatinib...