Objective To investigate the mechanisms of inhibitory effect of Erlotinib,an epidermal growth factor(VEGF) receptor inhibitor,on angiogenesis of pancreatic carncer.Methods①In a tube formation assay,Erlotinib(100?mol/L) was applied to the culture media and compared to the serum free media.The expression of vascular endothelial growth factor(VEGF) in BxPC-3 cells treated with Erlo- tinib at different concentrations(5,50,100,200?mol/L) was determined by RT-PCR.②The xeno grafts derived from BxPC 3 cancer cells were inoculated into the BALB/C nude mice.The mice were treated with either Erlotinib(100 mg/kg of Erlotinib oral lavage daily) or saline for four weeks.The vol- ume of the xenografts was measured and the tumor growth rate was calculated.The microvessel density (MVD) of tumor tissue was determined by immunohistochemistry with an antibody against factorⅧ. Results There were less endothelium cells and close hollow tubular structures in grlotinib treated group compared to the control group in the tube formation assay.The mean weight of xenografts in Erlotinib treated group[(0.397?0.550)g] was significantly lower than that in the control group[(1.570?1.060)g] with a inhibitary rate of 74.5%.The expression of VEGF mRNA in Ertotinib treated groups (=50?mol/L) were decreased comparing to the control group.The VEGF expression in xeno- grafts tumor tissues was also markedly down-regulated.The MVI) was significantly decreased in Erlotinib treated group( 1.86?0.43)than that in the control group (5.98?1.27,P

Objectives To investigate and evaluate the preventive and therapeutic effects of celecox ib,a selective cyclooxygenase-2(COX-2)inhibitor,on multiple colorectal adenorna and compare it with aspirin.Methods Ninty-six patients with colorectal multiple adenoma were randomly divided into A,B and C groups.Adenomas in all patients were removed with high-frequency eleetrocoagulation,electroexci- sion or argon plasma coagulation(APC)under colonoscopy.Then,group A were administered celecoxib 200 mg twice daily,group B aspirin 50 mg twice daily,group C served as control.Colonoscopy was per formed every 6 months in the first year,and every year in order to observe and evaluate the recurrence rate of adenoma and the side effects after the treatment.Results Twenty-seven patients in group A,26 pa- tients in group B and 27 patients in group C had completed the treatment.At the end of the treatment, on PP/ITT analysis,the cure rate of the eolorectal adenoma were 84.4%/100% ,78.1%/96.2% and 75.0%/88.9% in group A,B and C,respectively.During the first year of follow-up,there were 1 ,1 and 6 cases which were found recurrences of the adenomas in group A,B and C,respectively.The recurrence rates of coloreetal adenomas in group A(3.7%)and group B(4.0%)were significantly low er than that in group C(24.0%) (P＜0.05 and＜0.05,respectively).At the end of follow-up,the total recurrence rate of colorectal adenomas in group A(14.80%)and group B(19.2%)were significant- ly lower than that in group C(46.2%)(P＜0.05 and＜0.05).While the side-effective rate regroup A (3.3%)was significantly lower than that in group B(22.5%)(P＜0.05).Conclusions After re- section of the multiple colorectal adenomas,both the selective inhibitor of COX-2,celeeoxib and the non- selective inhibitor of COX-2,aspirin,may reduce its recurrence rate,but the former has a good tolerance and lower side-effects.

OBJECTIVETo investigate the differences about RNA interference (RNAi) technique which focuses on single or multiple sites to suppress colon cancer LoVo cell line's epidermal growth factor receptor (EGFR) mRNA and protein expression, induce cell apoptosis and enhance 5-fluorouracil (5-FU) sensitivity.

METHODSThe human colon cancer LoVo cells were transfected by liposome with pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2 expressive vectors which were established by p Genesil-1 plasmid and EGFR short hairpin RNA (shRNA) synthesized in vitro, then were selected for 4 weeks by using G418. Five groups were selected for the study: Group 1: the normal cultured LoVo cells; Group 2: the negative control plasmid HK; Group 3: pU6-EGFR-shRNA-1 plasmid vector; Group 4: pU6-EGFR-shRNA-2 plasmid vector; Group 5: pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2, half for each. The mRNA and protein expression were assessed using Real Time PCR and Western blot, the cell apoptosis was determined via flow cytometry, and the suppressive rate and IC(50) to LoVo cells by 5-FU of different concentrations and time points were carried out by using Cell Counting Kit-8 (CCK-8).

RESULTSExpression plasmids encoding shRNA were successfully established and transfected into the LoVo cells. In group 3, 4 and 5, the mRNA expression was decreased by (80.2 +/- 3.4)%, (81.3 +/- 2.8)% and (90.6 +/- 2.8)%, respectively, and protein expression was decreased by (74.1 +/- 4.0)%, (73.4 +/- 2.3)% and (90.4 +/- 3.3)%, respectively; meanwhile, cell apoptosis increased by (10.4 +/- 0.5)%, (10.1 +/- 0.4)% and (14.2 +/- 0.5)%, respectively. The IC(50) of 5-FU and cell suppressive rate analysis demonstrated that there were significant differences among group 5, groups 3 and 4, and groups 1 and 2, but there were no significant difference between group 1 and group 2, as well as group 3 and group 4.

CONCLUSIONSBoth pU6-EGFR-shRNA-1 and pU6-EGFR-shRNA-2 were capable of suppressing EGFR expression of LoVo cells, and therefore promoting apoptosis and increasing the cell toxicity of 5-FU. The targeting double combined sites RNAi technique was significantly better than single site interference. The new therapeutic modalities in the treatment of human colon cancer are suggested by this study.

Apoptosis ; genetics ; Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; therapy ; Combined Modality Therapy ; Fluorouracil ; pharmacology ; Genetic Therapy ; Humans ; RNA ; genetics ; RNA Interference ; Receptor, Epidermal Growth Factor ; biosynthesis ; genetics ; Transfection

OBJECTIVETo explore the efficacy of tumor-ablative individualized allogeneic hematopoietic stem cell transplantation for the treatment of patients with high risk/refractory leukemia.

METHODSFivety-seven patients with high risk/refractory leukemia were enrolled. Tumor-ablative individualized conditioning regimens included HDAra-C + Bu/Cy, Ara-C + Bu/Fludarabine, G-CSF primed HDAra-C + Bu/Cy, and FLAG followed by reduced-intensified BuCy. Overall survival (OS), disease free survival (DFS), graft versus host disease, infection and relapse post grafting were analyzed.

RESULTSFifty-six patients attained durable engraftment. The median follow-up duration was 17.5 (2 - 34) months. The 18 months probabilities of OS and DFS were (74.7 ± 6.1)% and (62.4 ± 6.7)%, respectively. In addition, the 18 months probabilities of OS and DFS in patients who attained complete remission (CR) before transplantation were (74.2 ± 7.1)% and (58.8 ± 8.1)%, respectively, while in those not attained CR were (77.0 ± 11.8)% and (72.7 ± 11.7)%, respectively. Twenty nine patients developed acute GVHD (aGVHD) (grade I in 18, grade II in 4, grade III in 2 and grade IV in 5). The probabilities of aGVHD was (50.9 ± 6.6)% by Kaplan-Meier curve analysis. The probabilities of grades 2-4 and grades 3-4 aGVHD were (19.3 ± 5.2)% and (12.3 ± 4.3)% respectively. Extensive chronic GVHD (cGVHD) was observed in 36 patients. The probabilities of cGVHD was (64.3 ± 6.4)% by Kaplan-Meier curve analysis. Cytomegaloviremia (CMV) was observed in 39 (68.42%) patients, hemorrhagic cystitis in 13 (22.8%) patients, fungous infection in 16 (28.07%) patients and bacterial infection in 38 (66.67%) patients. Relapse occurred in 14 patients (hematologic relapse in 11 and extramedullary relapse in 3), probabilities of relapse being (24.6 ± 5.7)%. The 17.5-month probability of relapse in patients who attained CR before transplantation was (28.1 ± 7.7)%, while in those not attained CR was (15.6 ± 10.2)%. Fifteen patients died (6 from hematological relapse, 5 from infection of bacterial and fungous, 4 from cGVHD) after 100 days.

CONCLUSIONTumor-ablative individualized allogeneic hematopoietic stem cell transplantation is a promising and safe choice for treatment of high risk/refractory leukemia, even with high leukemia burden.

Cytarabine ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; Transplantation Conditioning

OBJECTIVETo analyse the clinical features, diagnostic methods and risk factors of cytomegalovirus (CMV) enteritis after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSAnalysis was made on 24 cases of CMV enteritis after allo-HSCT in Beijing Daopei Hospital from Aug. 2007 to Jul. 2009, including clinical data, endoscopic diagnosis, histopathological and virological results, and the association between CMV enteritis with viremia and graft-versus-host disease(GVHD).

RESULTS87.5% of the patients were over 18 years old. The median time to diagnosis of CMV enteritis was 63 days after HSCT. The mucosal lesions in enteroscopic examination had no significant differences between CMV enteritis and gastrointestinal GVHD complicated with the enteritis. The methods used in diagnosis included histopathology (32.1%) and virology (92.9%). The copies of CMVDNA in mucosal samples greater than 10(5)/10(6) PBNC was better diagnosis. A number of risk factors were compared between the survival and death groups: type of transplant, conditioning regimen, the time span of ganciclovir prophylaxis therapy, grade II-IV GVHD before enteritis, the time of diagnosis as GVHD, using MP > or = 1 mg/kg to treat GVHD, the time between GVHD and enteritis, CMV viremia before enteritis, the time of diagnosis as enteritis, CMVDNA quantitation, and there were no any statistic differences.

CONCLUSIONCytomegalovirus enteritis should be carefully diagnosed by histopathology and virology through endoscopic examination. It is better to undertake pan-colon endoscopy as well as terminal ileum examination for more accurate diagnosis. PCR can significantly improve the detection rate. CMVDNA detection in patients' stool may be helpful to diagnosis, especially for those patients who can not stand the endoscopy examination.

Adolescent ; Adult ; Cytomegalovirus ; Cytomegalovirus Infections ; etiology ; DNA, Viral ; isolation & purification ; Enteritis ; etiology ; virology ; Female ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Risk Factors ; Young Adult

OBJECTIVETo report two de novo acute myeloid leukemia (AML) patients with t(11;22)(q23;q11.2) and summarize the clinical and biological characteristics.

METHODSBone marrow cells morphology, immunophenotype, chromosome karyotype, fluorescence in situ hybridization (FISH), PCR and gene sequencing were performed. Clinical manifestation and routine laboratory tests were analyzed.

RESULTSThe patients were diagnosed as AML-M₂ and AML-M₅ by morphology and immunophenotype results. Both patients carried t(11;22)(q23; q11.2) and one of them carried an additional chromosome abnormality. MLL-SEPTIN5 fusion transcript was identified in two patients by RT-PCR and sequencing. The two patients got hematologic complete remission after induction chemotherapy with daunorubicin, homoharringtonine, and cytarabine (DHA) or daunorubicin and cytarabine (DA). One of them relapsed and died during consolidation therapy with intermediate-dose cytarabine.

CONCLUSIONLeukemia with t(11;22)(q23;q11.2) chromosome translocation met the clinical and laboratory manifestations of AML. The MLL-SEPTIN5 fusion transcript was the distinctively biological etiology. Patients with t(11;22)(q23;q11.2) were vulnerable to relapse after conventional chemotherapy and had poor prognosis. Allogeneic hematopoietic stem cell transplantation should be recommended as early as possible.

Adult ; Chromosome Aberrations ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 22 ; Female ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; drug therapy ; genetics ; Male ; Prognosis ; Translocation, Genetic

This study was purposed to investigate the clinical features and related factors influencing prognosis of patients with severe intestinal graft-versus-host disease (siGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). 710 patients received allo-HSCT in Beijing Dao-Pei hospital from Jan 2007 to Jan 2011 were enrolled in this study. A total of 34 patients with siGVHD out of 710 patients were analyzed retrospectively, and the univariate analysis for related factors influencing prognosis were carried out by using SPSS 19.0 software. The results showed that the incidence of siGVHD was 4.79%, its medium occurrence time was 29 (18 - 210) days after allo-HSCT. 18 out of 34 patients with siGVHD received colonoscopy, among them 6 patients were complicated with viral enteritis. The deep ulcers could be found under colonoscope. Histopathologic examination revealed the viral inclusion bodies or positive viral antigen. Methylprednisolone (MP), cyclosporine A (CsA) or tacrolimus combined CD25 monoclonal antibody and oral budesonide were used for treatment of siGVHD. 29 out of 34 cases achieved complete response (CR) with CR rate of 85.29%, overall survival rate was 58.82% (20/34). 9 out of 29 cases achieving CR died of other complications. The univariate analysis of the related factor indicated the hyperacute GVHD is the adverse factor influencing overall survival of patients with siGVHD. It is concluded that early colonoscopy is an effective way for definitive diagnosis of siGVHD. The combined treatment including MP, CsA or tacrolimus, CD25 monoclonal antibody and oral budesonide shows a significant curative effects. Intensive treatment of complications in late period of GVHD can enhance the overall survival rate.
Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Gastrointestinal Tract ; Graft vs Host Disease ; diagnosis ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo evaluate the efficacy of salvaged allogeneic hematopoietic stem cell transplantation (allo-HSCT) for refractory/recurrent acute myeloid leukemia (AML).

METHODSA total of 45 patients with refractory/recurrent AML were enrolled from September 2006 to April 2010. The median blasts in bone marrow (BM) were 36% (20% to 92%) before conditioning. The donors were identical siblings (6) or unrelated ones (9) or haploidentical family members (30). Conditioning regiments were individualized according to patients' status, the regimen with high-dose cytarabine plus BuCy/CY was mostly used (20). The patients with impaired organ function received above regimen except using fludarabine instead of cyclophosphamide (16). FLAG followed by reduced-intensified BuCy was employed for the recipients with more than 40% blasts in BM (6) to reduce leukemia burden. TBI/CY or TBI/Fludarabine was used for the recipients with extramedullary infiltration of leukemia or multidrug resistant leukemia. G-CSF, MTX, NVT, Vm26, Acla or Thaltipa was added into conditioning regiments according to leukemia character.

RESULTSAll but 2 patients attained durable engraftment. The incidence of grade II to IV aGVHD and cGVHD were 34%, 59.1%, respectively. With median follow-up 30 (0.5 - 57) months, the relapse rate was 29.2%. Twenty-nine of 45 (60.2%) patients remained in complete remission since salvaged HSCT. Three-years disease-free survival and overall survival were 60.2% and 62.6%, respectively.

CONCLUSIONOur results indicated that the combination of salvaged HSCT with prophylactic immunotherapy might be a promising modality for treatment of refractory/recurrent AML, even with high leukemia burden.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Middle Aged ; Recurrence ; Survival Rate ; Transplantation Conditioning ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the differences of olfactory bulb (OB) volumes between younger and older, male and female, left-side and right-side in healthy middle and old-aged persons by MRI.

METHODSNinety five healthy middle and old-aged volunteers (male:female = 45:50) were divided into 2 groups, group one included persons aged from 50 to 69, group two included persons elder than 70. The left-side, right-side and both-side volumes of OB, the volumes of brain and the ratio of OB/brain were measured by MRI.

RESULTS(1) The left-side and both-side volumes of OB (x(-) +/- s), the volumes of brain [(39.89 +/- 8.7) mm(3), (81.70 +/- 16.8) mm(3) and (1281.86 +/- 140.2) cm(3)] in 50 - 69 years old group were respectively larger than those in >/= 70 years old group [(34.45 +/- 10.4) mm(3), (72.10 +/- 19.3) mm(3) and (1165.77 +/- 165.3) cm(3)], and the differences reached statistical significance (t were respectively 2.649, 2.449, 3.516, all P < 0.05). There were no significant differences of right-side OB volumes and the ratio of OB/brain between 50 - 69 years old group and >/= 70 years old group (t were respectively 1.904, 0.616, each P > 0.05). (2) The male's OB volumes of left-side, right-side and both-side, the brain volumes and the ratio of OB/brain were respectively larger than females', and the differences reached statistical significance (t were respectively 4.461, 3.630, 4.399, 3.800, 2.400, all P < 0.05). (3) The right-side OB volumes were larger than left-side's and significant differences were found in female group, 50 - 60 years old group and >/= 70 years old group (t were respectively 2.732, 2.117, 3.516, all P < 0.05). There were no significant differences of OB volumes between left-side and right-side in female (t = 2.649, P = 0.110). The ratio of right-side OB/brain were larger than the ratio of left-side's and the differences reached statistical significance (t = 3.183, P = 0.002).

CONCLUSIONSMRI could be used to measure the volume of OB. The older the people, the smaller the OB volumes. There was no influence of age on the ratio OB/brain. The OB volumes of right-side were larger than those of left-side. The OB volumes of male were larger than those of female.

Humans ; Magnetic Resonance Imaging ; Olfactory Bulb ; Smell

In order to understand the structural characteristics of squalene synthase genes in the triterpenoids biosynthesis pathway of Crataegus pinnatifida, the squalene synthase genes of C. pinnatifida was cloned and analyzed by bioinformatics and prokaryotic expression. Two squalene synthase genes CpSQS1 and CpSQS2 were cloned from C. pinnatifida fruit by RT-PCR. The ORF length of CpSQS1 and CpSQS2 were 1 239 bp and 1 233 bp respectively, encoding 412 aa and 410 aa respectively. CpSQS1 and CpSQS2 were predicted to be stable acidic proteins by online tools. The secondary structure was mainly composed of α-helix structure, and the tertiary structure was predicted by homology modeling. Structural functional domain analysis showed that 35-367 aa of CpSQS1 and CpSQS2 cDNA containing conserved trans-isoprenyl pyrophosphate synthase domains. Transmembrane domain analysis predicted that two transmembrane domains were founded in CpSQS1 and CpSQS2. The squalene synthase amino sequence of C. pinnatifida had higher homology with the known SQS of Salvia miltiorrhiza and Glycyrrhiza glabra. Phylogenetic tree analysis showed that CpSQS1 and CpSQS2 were clustered into one branch of MdSQS1 and MdSQS2, which were consistent with the phylogenetic rule. Prokaryotic expression vector pGEX-4 T-1-CpSQS1 and pGEX-4 T-1-CpSQS2 were transformed into Escherichia coli Transetta(DE3) for induction, and the target protein was successfully expressed at 65 kDa. The expression levels of CpSQS2 were significantly higher than that of CpSQS1 in three different developmental stages of C. pinnatifida. In this study, the full-length cDNA sequences of C. pinnatifida SQS1 and SQS2 were cloned and analyzed for the first time, which provided the foundation for further study on the metabolic pathway of C. pinnatifida triterpenoids.
Amino Acid Sequence ; Cloning, Molecular ; Crataegus/genetics* ; Farnesyl-Diphosphate Farnesyltransferase/genetics* ; Fruit/enzymology* ; Phylogeny ; Plant Proteins/genetics*