Study of long non-coding RNA Tug 1 expression in Egyptian colorectal adenocarcinoma patients

Purpose: Colorectal cancer (CRC) is one of the most fatal tumors worldwide. In Egypt, most CRC cases occur in individuals > 40 years old. TUG1 has been proved to be disrupted in different malignancies and may have a critical role in tumor progression, invasion, and metastasis. However, its role in CRC has not been adequately studied. Materials / Methods: Quantitative real-time polymerase chain reaction (PCR) was used to evaluate the expression levels of long non-coding RNA (LncRNA) taurine upregulated gene 1 (TUG1), in nonmetastatic and metastatic CRC tissues and adjacent noncancerous tissues as control. Results: LncRNA TUG1 expression was significantly upregulated in both nonmetastatic and metastatic CRC tissues, in comparison with the adjacent noncancerous tissue. It was found that TUG1 could have a possible prognostic role in CRC, by comparing the sensitivity and specificity of TUG1 with those of CEA and CA19-9. Conclusion: The results of the current study suggest that the LncRNA TUG1 participates in the malignant behaviors of CRC cells. (AU)

Construction of interferon alpha/beta receptor subunit 1 gene knockout Caco-2 cell line based on CRISPR/Cas9 system / 中国生物制品学杂志

@#Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

Bleeding per rectum during induction chemotherapy: Looking beyond the leukaemia – 2 case reports and review of literature

Synchronous malignancies involving acute leukemia and a solid organ are rare. Bleeding per rectum is a common manifestation of acute leukemia during induction chemotherapy and might mask the presence of synchronous colorectal adenocarcinoma (CRC). Here we present two rare cases of acute leukemia with synchronous CRC. We also review previously reported synchronous malignancies to investigate demographics, diagnosis, and treatment modalities. Management of these cases requires a multispecialty approach

Tissue factor expression in colorectal adenocarcinoma: association with angiogenesis and clinical and pathological aspects

Introduction: Tissue factor (TF) expression has been described in various neoplasms and was correlated with angiogenesis and metastases. Objectives: To describe TF expression in colorectal cancers, correlating it with microvessel density and clinical and pathological variables. Methods: Immunohistochemistry was used to determine TF expression and microvessel density. The Student t-test was used to compare high and low TF expression with microvessel density andwith age. The chi-squared test was used for other comparisons, and Kaplan-Meier curves were used for survival analyses. Results: Forty-three patients were operated with curative intent. Their mean age was 58.1±12.6 years old, and 62.8% were male. The rectum was the most common location (60,4%), and most tumors reached the serosa and peri-intestinal fat (72.1%). Lymph nodes were positive in 46.5%, and 72.1% of the tumors were moderately differentiated adenocarcinomas. Death occurred in 27.6±12.8months in 51.1% of the patients who had recurrence. Tissue factor expression was intense in 88.4%. There was a positive correlation between TF expression and microvessel density (p=0.02), and between TF and older age (p< 0.01). There was no correlation between TF expression and other variables (gender, histological type, penetration into the intestinal wall, and lymphatic and systemic metastases). Tissue factor expression did not correlate with survival. Conclusion: Tissue factor expression correlated with increased microvessel density and older age. Further studies are necessary to ascertain the clinical relevance of TF in colorectal cancer. (AU)

Clinicopathological Significance of Poorly Differentiated Clusters in Liver Metastatic Lesions of Colorectal Carcinoma / 肿瘤防治研究

Objective To investigate the clinicopathological significance of PDC in liver metastases and analyze the correlation of PDC between liver metastases and primary lesions. Methods Retrospective analysis of 72 matched cases of colorectal cancer with liver metastases was performed. The PDC in primary tumor and liver metastatic lesion was interpreted synchronously, and then the relationship between PDC in liver metastasis and clinicopathological parameters was analyzed based on the correlation of PDC between primary and metastatic lesions. In addition, PDC were interpreted in accordance with Uenos' standard. Results Among the 72 cases of liver metastasis of colorectal cancer, the number of G1, G2, and G3 graded by PDC was 28, 24, and 20, respectively. The PDC in liver metastatic lesion was correlated with tumor budding in liver metastatic lesion and PDC grade of primary lesion. No significant correlation with the size and number of liver metastatic lesion, the site, WHO grade, depth of invasion, lymph node metastasis, vascular invasion or tumor budding of the primary lesion was observed. Conclusion A positive correlation is found between liver metastasis of colorectal adenocarcinoma and PDC grade of primary tumor. Evaluating the PDC grade of primary tumor may provide a reference for the risk of liver metastasis.

Profiles of endoglin and vascular endothelial growth factor based on staging and histological grading of colorectal cancer and their relationship with bevacizumab therapy / Perfis da endoglina e do fator de crescimento endotelial vascular com base no estadiamento e na graduação histológica do câncer colorretal e sua relação coma terapia com bevacizumabe

Objective: The present study evaluated the profile of endoglin (CD105) and vascular endothelial growth factor (VEGF) based on staging and histopathological grading of colorectal cancer as well as their relationship with bevacizumab therapy. Methods: A total of 88 cases of colorectal adenocarcinoma were included in the present study. The levels of VEGF and CD105 protein were evaluated with enzymelinked immunosorbent assay (ELISA). Results: There was a significant difference in the level of CD105 (p=0.002) between metastases and non-metastases subjects, showing that CD105 was higher in metastases subjects (4.59 ng/ml). Therewas no significant difference in the level of VEGF based on the presence of metastasis (p=0.625). There was a significant difference in the levels of CD105 (p=0.038) and VEGF (p=0.010) between the subjects who received chemotherapy and those who did not. The CD105 level was higher in the subjects who received chemotherapy (4.43 ng/ml); conversely, the level of VEGF was lower in subjects who received chemotherapy (543.65 pg/ml). There was a statistically significant difference in the levels of CD105 (p=0.003) and VEGF (p=0.002) between subjects who received bevacizumab therapy and subjects who did not. The levels of CD105 were higher in subjects who received bevacizumab therapy (5.11 ng/ml); in contrast, the level of VEGF was higher in subjects who did not receive bevacizumab therapy (645.92 pg/ml). There was a significant positive correlation between CD105 and VEGF in subjects who did not receive bevacizumab (p<0.01). Conclusion: The results of this study support a hypothesis of "escape mechanism" in the failure of anti-angiogenesis therapy (anti-VEGF). (AU)

Objetivo: Este estudo avaliou o perfil da endoglina (CD105) e do fator de crescimento endotelial vascular (FCEV) com base no estadiamento e graduação histopatológica do câncer colorretal, assim como sua relação com a terapia com bevacizumabe. Métodos: No total, 88 casos de adenocarcinoma colorretal foram incluídos no presente estudo. Os níveis das proteínas FCEV e CD105 foram avaliados com ensaio imunoenzimático (ELISA, na sigla em inglês). Resultados Houve uma diferença significativa no nível de CD105 (p=0,002) entre indivíduos commetástases e semmetástases, que indicou que o nível de CD105 émais alto em indivíduos com metástases (4,59 ng/ml). Não houve diferença significativa no nível de FCEV com base na presença de metástases (p=0,625). Houve diferença significativa nos níveis de CD105 (p=0,038) e de FCEV (p=0,010) entre os indivíduos que receberam quimioterapia e os que não receberam. Encontrou-se um nível de CD105 mais alto nos indivíduos que submetidos a quimioterapia (4,43 ng/ml); Em contrapartida, encontrou-se um nível de FCEV mais baixo em indivíduos que submetidos a quimioterapia (543,65 pg/ml). Houve uma diferença estatisticamente significativa nos níveis de CD105 (p=0,003) e de FCEV (p=0,002) entre os indivíduos submetidos e não submetidos à terapia com bevacizumabe. Os níveis de CD105 foram mais elevados em indivíduos submetidos à terapia combevacizumab (5,11 ng/ml); em contraste, observou-se um nível de FCEV mais alto em indivíduos que não foram submetidos à terapia com bevacizumabe (645,92 pg/ml). Houve uma correlação positiva significativa entre CD105 e FCEV em indivíduos que não receberam bevacizumabe (p<0.01). Conclusão: Os resultados deste estudo corroboram a hipótese de "mecanismo de escape" na falha da terapia anti-angiogênica (anti-FCEV). (AU)

Resecciones colorrectales segmentarias en pacientes con colitis ulcerosa y displasia o adenocarcinoma / Segmental colectomy in patients with colitis-associated neoplasia or dysplasia

Resumen Introducción: El tratamiento estándar para los pacientes con colitis ulcerosa y displasia o adenocarcinoma de colon ha sido la proctocolectomía total, lo que conlleva una morbilidad y una reducción en la calidad de vida significativa. Materiales y Método: Se hace un análisis retrospectivo de 5 pacientes con colitis ulcerosa a los que se realiza una resección segmentaria por displasia o adenocarcinoma. Resultados: La mediana de edad al diagnóstico de colitis ulcerosa y de la neoplasia fue de 56 y 62 años respectivamente. El tiempo de evolución de la enfermedad fue de 1 a 13 años. La mediana de seguimiento postoperatorio fue de 57 meses apareciendo en uno de los pacientes un nuevo foco de displasia. Conclusiones: En determinados pacientes seleccionados, las resecciones segmentarias podrían ser una opción segura si tienen buen control de la enfermedad, escasa actividad inflamatoria, pocos años de evolución y que puedan realizar un adecuado seguimiento posterior.

Introduction: The historical management for patients with ulcerative colitis and displasia or adenocarcinoma associated was to perform a total proctocolectomy, what cause important morbidity and affect patient's quality of life. Materials and Method: A retrospective review about 5 patients with a segmental colectomy due to dysplasia or adenocarcinoma is done. Results: The median age at diagnosis of ulcerative colitis and cancer was 56 and 62 years respectively. Disease time evolution was between 1 and 13 years. The patients were followed up for a median of 57 months. During the follow-up evaluation, 1 patient was found to have dysplasia. Conclusion: Segmental colectomy could be a safe option in clinically stable patients, few years of diagnosis and with and a suitable follow up.

Growth pattern, tumor budding and CD8+T lymphocyte infiltration in colorectal adenocarcinoma invasive margins: association with clinicopathological features / 中国肿瘤临床

Objective: The aim of this study was to explore the correlation between the clinicopathological characteristics of colorectal adenocarcinoma and the growth pattern, tumor budding, and CD8+T lymphocyte infiltration in anterior invasive margins, and to assess their value as prognostic indicators. Methods: Paraffin embedded samples were collected from 126 patients with primary colorectal adenocarcinoma who underwent surgical resection in Zhangye People's Hospital Affiliated to Hexi College from January 2008 to December 2019. A total of 126 pathological sections were stained by immunohistochemistry. Anti-cytokeratin antibodies were used to mark tumor cell budding and anti-CD8 antibody markers T lymphocytes were evaluated. Results: In colorectal adenocarcinoma, infiltrative growth patterns and high-grade tumor budding in invasive margins were significantly associated with pathological stage of tumor size (pT) (P=0.029 and P<0.001, respectively), pathological stage of lymph node metastasis (pN) (P<0.001 and P=0.023, respectively) and vessel infiltration (P<0.001 and P<0.001, respectively). Furthermore, high-grade CD8+T lymphocyte infiltration was associated with the absence of lymph node metastases (P=0.050). Conclusions: Infiltrative growth patterns and high-grade tumor budding in colorectal adenocarcinoma invasive margins were correlated with patient prognosis. Importantly, these two features are easily detectable (with the help of pan-cytokeratin immunohistochemistry staining), in a reproducible manner. Therefore, we propose that they could be used as prognostic indicators in colorectal adenocarcinoma patients.

Expression and significance of Notch3, DLL1 and CD133 in colorectal adenocarcinoma / 重庆医学

Objective To investigate the expression of Notch3,DLL1 and CD133 in human colorectal adenocarcinoma and its clinical pathology meaning.Methods Immunohistochemical staining was used to detect the expression of Notch3,DLL1 and CD133 in 12 cases of normal colorectal mucosa tissue,30 cases of colorectal adenoma tissue and 50 cases of colorectal adenocarcinoma tissue,and the relationship between them and clinicopathological data wereanalyzed.Results The positive rates of Notch3,DLL1 and CD133 were 64.0 ％ (32/50),68.0 ％ (34/50) and 54.0 ％ (27/50) in colorectal carcinoma tissues,respectively,which were remarkably higher than those in colorectal adenoma tissue(26.7 ％,33.3％％,36.7 ％)and those in normal colorectal mucosa tissue (8.3％,16.7％,8.3％) (P＜0.05).There was no significant difference between the adenoma group and the normal group(P＞0.05).The expression levels of Notch3,DLL1 and CD133 were not correlated to age,gender,tumor location,degree of differentiation and the tumor size,except lymph node metastasis,in addition,the expression of Notch3 and DLL1 were also associated with Dukes staging,and The expression of DLL1 were also associated with tumor infiltration depth (P＜0.05).The expression of Notch3 protein was positively related to that of DLL1 protein(r=0.478,P=0.000).Conclusion The positive expression rates of Notch3,DLL1 and CD133 in colorectal adenocarcinoma tissue are remarkably higher than those in the normal colorectal mucosa and colorectal adenoma tissue,which prompts that the high expression of Notch3,DLL1 and CD133 may all participate in the process and metastasis of colorectal adenocarcinoma.Moreover Notch3 signalling pathway may act through the cancer stem cells and eventually regulate the initiation and development of colorectal adenocarcinoma.

Cáncer colorectal en la edad pediátrica: reporte de un caso

El cáncer colorrectal (CCR) constituye una dolencia extremadamente infrecuente en niños. Es más excepcional cuando ocurre en niños con neurofibromatosis tipo 1, que es considerada como factor de riesgo para el desarrollo de tumores malignos. Se reporta el caso de un niño de 8 años con CCR metastático de alto grado. El paciente presentó diarrea crónica muco-sanguinolenta, rectorragia y pérdida de peso. Su examen reveló manchas café con leche diseminadas sobre la espalda, pelvis y miembros inferiores. En la colonoscopia se visualizó una masa polipoide, estenosante, pediculada, ulcerativa, localizada a 35 cm del margen anal. La TAC reveló una metástasis hepática. Se realizó una colectomía amplia y el estudio histológico demostró 23 pólipos, 112 ganglios reaccionales y un tumor con componente mucinoso. Recibió quimioterapia y lobectomía hepática. No hay protocolo específico para el tratamiento de esta afección en niños. El pronóstico depende principalmente del carácter agresivo del tumor, de su estadificación y del lugar en el que se desarrolla.

Colorectal cancer (CRC) is an extremely rare condition in children. It is more exceptional when it oc-curs in children with neurofibromatosis type 1, which is considered a risk factor for the development of malignant tumors. We report the case of an 8-year-old child with high-grade metastatic CRC. The patient had chronic muco-bloody diarrhea, rectal bleeding and weight loss. His examination revealed cafe-au-lait spots scattered on his back, pelvis, and lower limbs. A polipoid, stenosing, pediculated, ul-cerative mass located 35 cm from the anal margin was visualized by colonoscopy. CT revealed hepatic metastasis. An ample colectomy was performed and the histology showed 23 polyps, 112 reactive lymph nodes and one tumor with a mucinous component. The patient received chemotherapy and liver lobectomy. There is no specific protocol for the treatment of this condition in children. The prog-nosis depends mainly on the aggressive nature of the tumor, its staging and the place where it develops.

Expression of USP10 in colorectal adenocarcinoma and analysis of bioinformatics / 临床与实验病理学杂志

Purpose To investigate the expression and significance of USP10 protein and mRNA in normal colorectal mucosa and colorectal adenocarcinoma,and to analyze the cause of the disorder.Methods 99 cases of colorectal adenocarcinoma and 83 cases of normal intestinal mucosa tissue were selected.Using tissue microarray and immunohistochemistry the expression of USP10 protein was detected,and the relationship was analyzed between USP10 protein and clinical pathological parameters or prognosis survival time.The expression of USP10 mRNA was analyzed by GEO datesets.Some miRNAs that down-regulate the expression of USP10 protein were screened by bioinformatics methods.The expression of USP10 protein and miR-149 in colorectal cancer cell lines were detected by Western blot and real-time quantitative PCR.Results The positive rate of USP10 protein in normal intestinal mucosa tissues was 71.08％(59/83),which was significantly higher than that in colorectal adenocarcinoma tissues (53.54％,53/99,P =0.015).No correlation were proved between USP10 protein expression and clinical pathological parameters or survival time (P ＞ 0.05).The expression level of USP10 mRNA in colorectal adenocarcinoma was 1.07 ～ 1.45 times that were higher than that of normal intestinal mucosa,which showed that the down-regulation of USP10 protein was at the post-transcriptional level.The program predicted a putative highly-conserved binding site in the USP10 mRNA 3'UTR for miR-149 which was up regulated in colorectal adenocarcinoma tissues.In addition,the expression of miR-149 was negatively correlated with the expression of USP10 protein in colorectal cancer cell lines.Conclusion The down-regulation of USP10 protein which occurs at the post-transcriptional level is closely related to the pathogenesis of colorectal adenocarcinoma.The high expression of miR-149 may be one of the factors that negatively regulate the expression of USP10 protein.

Anticancer property of berberine in human colorectal adenocarcinoma / 中草药

Objective: To illustrate the anticancer property induced by berberine, which is extracted from Chinese medicine Coptidis Rhizoma. Methods: Human colorectal adenocarcinoma cells, HCT-15, were treated with different concentration of berberine. Cells were then observed under the optical microscope to analyze the morphological changes; CCK-8 assay was used to detect the inhibition of cell proliferation of HCT-15; Immunoblotting was used to detect the changes of autophagy and apoptosis related markers. Athymic mice injected with HCT-15 cells were used to detect the pharmacological activity of berberine in vivo. Results: After berberine treatment, with the increased dose of berberine, the cell number reduced, cell spacing increased, and cell shape shrunk. The IC50 for berberine treatment in HCT-15 cell was 50 μmol/L. Autophagy and apoptosis markers were enhanced after treatment of 0, 50, and 100 μmol/L of berberine. The data further showed that enhanced autophagy and apoptosis were the main contributors for inhibiting cell proliferation. Immunoblotting analysis showed that berberine induced HCT-15 cells to autophagy and apoptosis mainly through p38 pathway. Athymic mice study showed that berberine treatment could decrease the volume of tumor in vivo, and the tumor inhibitory rates by berberine at 0, 50, and 100 mg/kg were 0, 32% and 74%, respectively. Conclusion: Berberine inhibits the HCT-15 cell proliferation, mainly by p38 pathway and enhances autophagy and apoptosis. Berberine can effectively inhibit tumor growth in vivo.

Aurora Kinase A Is a Prognostic Marker in Colorectal Adenocarcinoma

BACKGROUND: Aurora kinase A (AURKA), or STK15/BTAK, is a member of the serine/threonine kinase family and plays important roles in mitosis and chromosome stability. This study investigated the clinical significance of AURKA expression in colorectal cancer patients in Korea. METHODS: AURKA protein expression was evaluated by immunohistochemistry in 151 patients with colorectal adenocarcinoma using tissue microarray blocks. We analyzed the relationship between clinicopathological characteristics and AURKA expression. In addition, the prognostic significance of various clinicopathological data for progression-free survival (PFS) was assessed. Also we evaluated copy number variations by array comparative genomic hybridization and AURKA gene amplification using fluorescence in situ hybridization in colorectal carcinoma tissues. RESULTS: AURKA gene amplification was found more frequently in the 20q13.2–13.33 gain-positive group than the group with no significant gain on the AURKA-containing locus. AURKA protein expression was detected in 45% of the cases (68/151). Positive staining for AURKA was observed more often in male patients (p = .035) and distally located tumors (p = .021). PFS was shorter in patients with AURKA expression compared to those with low-level AURKA expression (p < .001). Univariate analysis revealed that AURKA expression (p = .001), age (p = .034), lymphatic invasion (p = .001), perineural invasion (p = .002), and TNM stage (p = .013) significantly affected PFS. In a multivariate analysis of PFS, a Cox proportional hazard model confirmed that AURKA expression was an independent and significant prognostic factor in colorectal adenocarcinoma (hazard ratio, 3.944; p < .001). CONCLUSIONS: AURKA could serve as an independent factor to predict a poor prognosis in Korean colorectal adenocarcinoma patients.

Anatomical hepatectomy for liver metastasis from rectal adenocarcinomapresenting with intrabiliary extension: a case report / Journal of Rural Medicine

<p>Liver metastases from colorectal carcinoma commonly form nodular lesions in the liverparenchyma. We report a case of liver metastasis from rectal adenocarcinoma that extendedpredominantly into the bile duct. A 62-year-old Japanese man underwent low anteriorresection for rectal adenocarcinoma 9 years ago. Approximately 3 years later, he underwentradiofrequency ablation therapy for a metastatic liver tumor. Nine years after surgery, atumor in liver segment III exhibiting intrabiliary extension was discovered; it wasunclear if this was a metastatic liver tumor or intrahepatic cholangiocarcinoma.Accordingly, we performed a left hepatectomy with lymph node dissection. The tumor wasnegative for cytokeratins 7 and 20, and was histologically similar to the primary rectaladenocarcinoma; it was diagnosed as rectal carcinoma metastasis. The patient has survivedfor 3 years after the hepatic surgery, for 9 years after radiofrequency ablation therapy,and for 12 years after the primary surgery. This case shows that liver metastasis fromcolorectal carcinoma can present as a predominantly intrabiliary growth that mimicsintrahepatic cholangiocarcinoma on imaging. Moreover, our case provides evidence for thesuperiority of anatomical hepatectomy over partial hepatectomy for metastatic liver tumorswith intrabiliary growth arising from rectal adenocarcinomas.</p>

Expression of CDX-2 and Ki-67 in different grades of colorectal adenocarcinomas.

Background: CDX2 is a caudal homeobox gene essential for intestinal differentiation and is specifi cally expressed in colorectal adenocarcinomas. Its role in colorectal carcinogenesis is not fully elucidated. Aims and Objectives: To study the expression pattern of CDX2 and Ki-67 in different grades of colorectal adenocarcinomas and to observe the relationship of their staining patterns in various tumor stages and to look for correlation if any, between Ki-67 labeling index (Ki-67 LI) and CDX2 expression. Materials and Methods: A total of 74 cases were enrolled. Detailed clinical profi le, peroperative fi ndings, histological grading and staging were noted. Immunohistochemistry for CDX2 and Ki-67 was done, and Ki-67 LI was calculated. CDX2 staining was graded semiquantitatively, and statistical analysis was done. Result: Age of presentation ranged from 20 to 75 years, and the male:female ratio was 1.83:1. There were 8, 47 and 13 cases of well, moderate and poorly differentiated adenocarcinomas, respectively. The mean Ki-67 LI of well, moderate and poorly differentiated adenocarcinomas were 14.25, 31.34 and 43.08 respectively, and their difference was statistically signifi cant, correlation was also noted with stage. CDX2 expression appeared to be stronger in poorly differentiated cases, but there was no signifi cant difference in its expression in the different grades and stages. There was no correlation between Ki-67 LI and CDX2 immunostaining pattern. The lymph node metastasis showed CDX2 positivity in all the cases. Conclusion: Expression of CDX2 does not signifi cantly change with the grade of colorectal adenocarcinomas. However, it is an important diagnostic marker in metastatic colonic lesions. The Ki-67 LI, on the other hand, showed a strong correlation with histopathological grades.

Expression of enhancer of zeste homolog2 in colorectal adenocarcinoma and its significance / 肿瘤研究与临床

Objective To investigate the expression of the enhancer of zeste homolog 2 (EZH2) gene and its significance in colorectal adenocarcinoma.Methods Immunohistochemistry and Western blot was used to assess the expression of EZH2 in human colorectal adenocarcinoma tissues,colorectal adenoma tissues and non-cancerous adjacent colorectal tissues.The relationships between EZH2 and each clinical pathology parameter were analyzed.Results The results of immunohistochemical trail showed that the expression rates of EZH2 in colorectal adenocarcinoma,colorectal adenoma and non-cancerous adjacent colorectal tissues were 80.56 ％ (87/108),62.5 ％ (25/40) and 5.00 ％ (2/40),respectively (P ＜ 0.05).Western blot revealed that the expression level of EZH2 in colorectal adenocarcinoma tissues,colorectal adenoma tissues and non-cancerous adjacent colorectal tissues level 0.549±0.145,0.283±0.023 and 0.107±0.022,respectively.The level in colorectal adenocarcinoma tissues (0.549±0.145) and colorectal adenoma (0.283±0.023) was significantly higher than that in non-cancerous adjacent colorectal tissues (0.107±0.022).Compared with colorectal adenoma tissues,level in colorectal adenocarcinoma tissues was significantly higher.There were significant differences among the three groups (F =20.113,P ＜ 0.05).The ratio of high expression level of EZH2 in colorectal adenocarcinoma tissues was closed related with tumorgenesis,differentiation,TNM staging and lymphatic metastsis (all P ＜ 0.05).However,no correlation was revealed between EZH2 expression and the age,gender (both P ＞ 0.05).Conclusion The expression of EZH2 may be associated with the tumorgenesis invasion,metastasis and progression of colorectal adenocarcinoma.

Expression and clinical significance of VEGF and Id-1 in ccervical cancer tissues / 中国基层医药

Objective To investigate the expression VEGF and Id -1 and its possible relationship with the c1inical pathological features of cervical cancer .Methods The expression of VEGF and Id -1 were examined in cer-vical cancer and normal tissues by immunohistochemical SP technique .Results The positive expression rate of VEGF in cervical cancer was 80.00%(48/60),which was significantly higher than that in normal tissues 6.67%(2/30)(P<0.01);The positive expression rate of Id -1 protein in cervical cancer was 73.34%(44/60),which was significantly higher than that in normal tissues 10.00%(3/30)(P<0.01);The expressions of VEGF and Id -1 were positively correlated with the degree of differentiation ,vascular invasion,and FIGO stage(P<0.05),and while was not significantly associated with age ,tumor size,lymph node metastasis ,and pathologic types .Expression of VEGF was positively related with expression of Id -1 in cervical cancer tissues(r=0.642,P=0.000).Multiple regression anal-ysis showed that:VEGF,Id -1,differentiation,vascular invasion,and FIGO were independent prognosis factors for cervical cancer .Conclusion The expressions of VEGF and Id -1 are correlated with the development of cervical cancer.Detection of the expression of VEGF and Id -1 in cervical cancer can predict the metastasis and prognosis , which can provide the basis for clinical targeted therapy .

Pharmacokinetic aspects and in vitro-in vivo correlation potential for lipid-based formulations

Lipid-based formulations have been an attractive choice among novel drug delivery systems for enhancing the solubility and bioavailability of poorly soluble drugs due to their ability to keep the drug in solubilized state in the gastrointestinal tract. These formulations offer multiple advantages such as reduction in food effect and inter-individual variability, ease of preparation, and the possibility of manufacturing using common excipients available in the market. Despite these advantages, very few products are available in the present market, perhaps due to limited knowledge in the in vitro tests (for prediction of in vivo fate) and lack of understanding of the mechanisms behind pharmacokinetic and biopharmaceutical aspects of lipid formulations after oral administration. The current review aims to provide a detailed understanding of the in vivo processing steps involved after oral administration of lipid formulations, their pharmacokinetic aspects and in vitro in vivo correlation (IVIVC) perspectives. Various pharmacokinetic and biopharmaceutical aspects such as formulation dispersion and lipid digestion, bioavailability enhancement mechanisms, impact of excipients on efflux transporters, and lymphatic transport are discussed with examples. In addition, various IVIVC approaches towards predicting in vivo data from in vitro dispersion/precipitation, in vitro lipolysis and ex vivo permeation studies are also discussed in detail with help of case studies.

Preliminary screening for microsatellite instability and loss of heterozygosity in the deleted in colorectal cancer (DCC) gene among Filipino patients with colorectal adenocarcinoma

OBJECTIVE: This study aimed to detect the presence of microsatellite (MSI) and loss of heterozygosity (LOH) of the Deleted in Colorectal Cancer (DCC) gene in normal and tumor tissues of Filipino colorectal cancer patients and examine its correlation with age, gender, tumor grade, tumor stage and site of lesion.METHODS: Paired frozen normal and tumor tissues from thirtynine (39) patients with colorectal adenocarcinoma were used by polymerase chain reaction (PCR). Single strand conformation polymorphism - polyacrylamide gel electrophoresis (SSCP - PAGE) was used to determine MSI and restriction fragment length polymorphism (RFLP) was used to study LOH.RESULTS: Based on our data, out of the 39 patients, 10 showed LOH of the DCC gene using the LOH markers VNTR, M2 and M3, while no MSI was detected in the samples using the MSI markers BAT25 and BAT26. Correlation with clinicopathological characteristics showed that there is significance for the site of lesion. The LOH has correclation with tumor samples from the colon but not with those from the rectum.CONCLUSION: Preliminary screening for MSI and LOH of the DCC gene shows that occurrences of colorectal cancer among Filipino patients can be correlated with LOH of the DCC gene with colorectal cancer in a Filipino sample population.

Integrated gene expression signature analysis of prognosis and drug-resistance in patients with serrated colorectal adenocarcinoma / 第二军医大学学报

Objective: To conduct a integrated analysis of gene expression signature and gene expression profile, so as to provide reference for the prognosis and drug-resistance of serrated colorectal adenocarcinoma (SCA). Methods: We downloaded four gene expression datasets (GSE14333, GSE17538, GSE33113, and GSE37892) of colorectal carcinoma with the follow-up survival data from GEO database, and then integrated them into a entire expression profile (n=600) with batch adjustment and gene expression value extraction. An SCA gene signature and the corresponding expression profile were integrated with the previous expression dataset. Using the gene signature score model we assigned score to each patient in the gene expression dataset and classified the patients into serrated, transitional, or conventional subtypes. Kaplan-Meier analysis and Cox model were used to compare the risks of cancer recurrence between three subtypes of colorectal carcinoma. Gene signature model were also used to generate the score for each patient in the datasets associated with alleviative therapy of colorectal cancer (GSE28702, GSE5851). The SCA drug-resistance was analyzed by observing the therapeutic effective group and non-effective group. Results: According to cut-off value of CRC subtypes, 600 patients in the combined dataset were classified as 50 with serrated subtype, 126 with transitional subtype, and 424 with conventional subtype. Survival analysis showed the serrated and transitional subtypes had very similar scores to predict patient survival, and they were also independent risk factors for postoperative recurrence. When comparing serrated and conventional subtypes, multivariate Cox model analysis indicated the patients with serrated subtype was an unfavorable independent risk factor for prognosis (AHR=1.792, 95%CI 1.011-3.177). Responders to cetuximab treatment had significantly higher signature scores than non-responders (P=0.017), while responders to FOLFOX treatment had similar signature scores with the non-responders. Conclusion: Serrated subtype is an independent risk factor of postoperative recurrence in SCA patients, and is related to the treatment with cetuximab.