Objective: To observe the expression of phosphorylated mammalian target of rapamycin(p-mTOR), epidermal growth factor receptor(EGFR), growth factor receptor-bound protein 2(Grb2) and vascular endothelial growth factor(VEGF) in human colorectal cancer tissues and to explore their roles in the carcinogenesis of colorectal cancer. Methods: Tissue microarray containing 185 colorectal cancer tissues was constructed and the expression of EGFR, Grb2, p-mTOR and VEGF in the colorectal cancer tissues and the corresponding adjacent tissues was examined by immunohistochemistry methods. The relationship between their expression with the clinicopathological characteristics such as age, sex, invasion depth, lymphatic metastasis, clinical stage and differentiation degree was analyzed. Results: EGFR, Grb2, p-mTOR and VEGF were scarcely expressed or absent in the corresponding adjacent tissues; their positive rates in the colorectal caner tissues were 21.1%, 44.9%, 42.2% and 54.1%, respectively, which were significantly higher than those in the corresponding adjacent tissues (P<0.05). The expression of EGFR, Grb2, p-mTOR and VEGF was not correlated with the patients' sex, age and differentiation degree of cancer. Overexpression of EGFR was found significantly associated with the invasion depth and clinical stage of cancer(P<0.05); and overexpression of p-mTOR and VEGF was significantly associated with lymphatic metastasis, invasion depth and clinical stage(P< 0.05). There was a correlation between every two of the four proteins (r=0.245-0.567, P<0.05). Conclusion: Over-expression of EGFR, Grb2, p-mTOR and VEGF is closely associated with the development and progresssion of colorectal cancer, and they may be worth further studying as new targets for the molecular target therapy of colorectal cancer.

Objective: To explore the roles of mammalian target of rapamycin (mTOR) and the activated mTOR (phosphorylated mTOR, p-mTOR) in the development and progression of colorectal cancer, and to discuss the clinical significance. Methods: The expression of mTOR and p-mTOR in 185 coLorectal cancer specimens and the corresponding adjacent tissues were evaluated by tissue microarray and immunohistochemistry,and the relationship between the expression and the age, sex, invasion depth( T stage) ,lymph metastasis, TNM stage and differentiation degree was analyzed. Results1 Diffused expression of mTOR and hardly any expression of p-mTOR were found in the adjacent tissues. The expression of mTOR and p-mTOR was obviously stronger in the colorectal cancer tissues compared with that in the adjacent tissues. The over-expression rates of mTOR and p-mTOR in colorectal cancer were 45. 9% and 42. 2%, respectively. There was no significant correlation of mTOR and p-mTOR over-expression with age, sex( P> 0. 05); the over-expression of mTOR was correlated with the differentiation degree (P<0. 05) ,but not with the invasion depth (T stage) ,TNM stage,or lymph metastasis (P>0. 05). The correlation of p-mTOR over-expression with the invasion depth (T stage) ,lymph metastasis and TNM stage was significant (P<0. 05) ,but that with the differentiation degree was not significant (P>0. 05). Conclusion:Over-expression of p-mTOR is closely associated with the malignant phenotype of colorectal cancer. It is also indicated that p-mTOR may be involved in the development and progression of colorectal cancer.

Cerebral Hemorrhage ; etiology ; Cerebrospinal Fluid Rhinorrhea ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Hemorrhage

Objective To study role of MDC/CCI22-CCR4 axis in mouse milky spots with peritoneal carcinomatosis of gastric cancer. Methods We examined the expression of CCR4 in 615 Mouse gastric cancer cell (MFC) lines by RT-PCR and Western-blot; Peritoneal metastasis model on the 615mouse was established by intraperitoneal injection of 0.2 ml MFC cells(1×104 cells). Dil fluorescence was used to observe the transfer process and section of MFC. Immunohistochemistry was conducted to detect the expression of CCR4 and CCL22 in omental milky spot; the structure of Milky spot was observed by scanning electron microscopy. Mice were randomly divided into 2 groups, namely, the saline control group (received saline) and MFC group. The concentration of CCL22 in ascitic fluid was measured in the 615 mice injected MFC after 6,8,10 days and in the saline group. Results MFC first metastasizes to the milky spot on the omentum, the expression of CCR4 and CCL22 were observered in the milky spot. The surface layer cells in milky spot consisted of discontinuous mesothelial cells and mainly macrophages and lymphocytes. The average value of CCL22 was 43 pg/ml and 364 pg/ml respectively in saline control group and MFC group.Conclusions MDC/CCL22-CCR4 axis plays an important role in the development of peritoneal carcinomatosis in mouse gastric cancer.

OBJECTIVE:To investigate the effect of different doses of oxaliplatin on the efficacy and safety and related index-es of patients with hepatocellular carcinoma(HCC)after transcatheter arterial chemoembolization(TACE). METHODS:100 HCC patients were randomly divided into control group (50 cases) and observation group (50 cases). After TACE,control group re-ceived arterial infusion chemotherapy of 40 mg/m2 oxaliplatin,once a day+20 mg Epirubicin hydrochloride for injection,once a day,with little lipiodol. Observation group received arterial infusion chemotherapy of 80 mg/m2 oxaliplatin (the same usage with control group)+Epirubicin hydrochloride for injection(the same dosage with control group),with little lipiodol. Alanine aminotrans-ferase (ALT),aspartate aminotransferase (AST),total bilirubin (TBIL),white blood cell count (WBC) and alpha-fetoprotein (AFP) in 2 groups after 3 d treatment were observed,and the total overall survival (OS) and the incidence of adverse reactions were followed-up. RESULTS:Abdominal pain,incidences of nausea and vomiting,ALT,AST and TBIL in observation group were significantly higher than control group,WBC was significantly lower than control group,the differences were statistically sig-nificant (P<0.05). There were no significant differences in fever,hair loss,incidence of peripheral neurotoxicity,OS in 1 year and 3 years,and AFP in 2 groups(P>0.05). CONCLUSIONS:Compared with 40 mg/m2 oxaliplatin,80 mg/m2 can reduce the in-cidence of abdominal pain,nausea,vomiting,but other effects are poor than 40 mg/m2,and it can not prolong the survival time of patients.

OBJECTIVE:To observe the clinical efficacy of entecavir combined with intervention in the treatment of liver cancer patients with HBV DNA-negative hepatitis B. METHODS:100 liver cancer patients with HBV DNA-negative hepatitis B were random-ly divided into observation group and control group,50 cases in each group. Control group received intervention,percutaneous punc-ture of femoral artery,then injected Pirarubicin hydrochloride for injection + lipiodol in parent artery until blood stagnation,and also conventional liver protection therapy was conducted;observation group additionally received 1 mg Entecavir tablet,qd,for continuous 6 months. Clinical efficacy,HBV DNA quantification,Child-Pugh score and liver function indexes in 2 groups were compared. RE-SULTS:The total effective rate in observation group was 44.0%,which was significantly higher than control group(26.0%),the dif-ference was statistically significant(P<0.05). There were no significant differences in HBV DNA quantification,Child-Pugh score and liver function indexes between 2 groups(P>0.05). After treatment,HBV DNA quantification,Child-Pugh score,fetoprotein,alanine aminotransferase, total bilirubin and aspartate aminotransferase significantly decreased in observation group,the differences were statis-tically significant compared with control group (P<0.05). CONCLUSIONS:Entecavir combined with intervention can obviously im-prove the clinical efficacy and liver function indexes of liver cancer patients with HBV DNA-negative hepatitis B.

DEK protein's carboxy-terminal DNA-binding region(CBD)is a newly found DNA-binding domain of DEK,which contains several phosphorylation sites and has a close correlation with DEK protein's function in vivo and in vitro.Using prokaryotic expression system,the peptide of DEK protein's carboxy-terminal DNA-binding region(CDB)was expressed and purified.In detail,the CDB DNA fragment was constructed into pET-30a(+)vector,and E.coli BL21(DE3)competent cells were used as host cells.The fusion protein His-CBD was expressed by induction of IPTG and purified by Ni-NTA agarose.The result of SDS-PAGE showed that the molecular weight of the purified protein was about 10.7kDa.Electrophoretic mobility shift assay(EMSA)indicated that DEK-CDB prefered to bind to supercoiled form of DNA in vitro,it had similar character to the binding of whole length DEK protein with DNA.This suggested that the carboxy-terminal DNA-binding region of DEK protein might function on the binding of DEK protein to DNA partly.

Objective To study the characteristics of mesenchymal stem cells(MSCs)derived from chronic myelogenous leukemia(CML)patients' bone marrow and examine their hematopoiesis support in vitro.Methods MSCs from CML were obtained and cultured.Immunophenotype and in vitro differentiation capacities were investigated.Moreover,the Ph chromosome and bcr/abl gene of CML derived MSCs were detected.The expression of cytokines was detected by RT-PCR,and hema- topoiesis support of MSCs in vitro was detected by long-term bone marrow culture and the methylcel- lulose progenitor assay.Results CML derived MSCs showed a typical fibroblast like morphology. They were positive for CD29,CD44 and CD105,while negative for CD14,CD31,CD45,CD34 and HLA-DR.Under suitable conditions,CML derived MSCs could differentiate into osteoblasts and adi- pocytes.Further,CML derived MSCs showed a normal choromosome,and did not express bcr/abl gene.At last,they expressed hematopoiesis cytokines and possessed ability of hematopoiesis support in vitro.Conclusion There are MSCs with multidirectional differentiation potential in CML bone mar- row and they possess the ability of hematopoiesis support.

Adult ; Humans ; Joint Dislocations ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Spinal Fractures ; surgery

In order to establish the optimum condition for purifica tion of the nucleoprotein(NP) of rabies virus by immunoaffinity chromatography, the efficient and non-denaturative eluents(Mg-el) was obtained by using ELISA elution model; furthermore, it didn't damage the activity of NP. Two kind of NPs , expressed by recombinant vaccinia virus (rVac-N) and recombinant baculovirus (BRN), were purified by a Sepharose CL 4B column and a 2C12- Sepharose 4B colum n. By Western-blot and SDS-PAGE, high purity and good antigenical intact NPs w ere identified. The purified ribonucleoprotein (RNP) of rabies virus 5aG strain was also obtained. After immunized with NP and RNP, mice developed a strong anti -nucleoprotein response and were protected against a lethal challenge of rabies virus CVS strain. There were not difference been observed among the mice immuni zed with different purified protein. These data indicate that the NPs are antige nical and immunogenical comparable to the authentic rabies RNP and therefore pre sent a potential source of an effective ,safe and economical subunit vaccine.