RESUMO
Objective: To explore the feasibility of clinical factors to predict the pathological complete response after neoadjuvant chemoradiotherapy in rectal cancer. Methods: A retrospective analysis was performed on clinical factors of 162 patients with rectal cancer, who underwent neoadjuvant chemoradiotherapy in the General Hospital of People's Liberation Army from January 2011 to December 2018.According to the postoperative pathological results, the patients were divided into pathological complete response (pCR) group and non-pathological complete response group (non-pCR group) to check the predictive clinical factors for pCR. Results: Twenty-eight cases achieved pCR after neoadjuvant chemoradiation (17.3%, 28/162). Univariate analysis showed that patients with higher differentiation (P=0.024), tumor occupation of the bowel lumen≤1/2 (P=0.006), earlier clinical T stage (P=0.013), earlier clinical N stage (P=0.009), the time interval between neoadjuvant chemoradiotherapy and surgery>49 days (P=0.006), and maximum tumor diameter≤5 cm (P=0.019) were more likely to obtain pCR, and the differences werestatistically significant. Multivariate analysis showed that tumor occupation of the bowel lumen≤1/2 (P=0.01), maximum tumor diameter≤5 cm (P=0.035), and the interval>49 days (P=0.009) were independent factors in predicting pCR after neoadjuvant therapy. Conclusion: Tumor occupation of the bowel lumen, maximum tumor diameter, and the time interval between neoadjuvant chemoradiotherapy and surgery can predict the pCR in rectal cancer.
Assuntos
Quimiorradioterapia , Terapia Neoadjuvante , Neoplasias Retais/terapia , Humanos , Estadiamento de Neoplasias , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Objective: The aim was to investigate the genotype distribution of two major epitopes of large surface protein (PreS1) of hepatitis B in Chinese patients and to explore the association between the genotypes of these two epitopes, and to determine whether PreS1 full-length genotype could be revealed according to the polypeptide sequence of key epitopes. Methods: HBV DNA was extracted from the serum of patients for PCR amplification. 278 samples amplified successfully were sequenced and compared with the known HBV sequences in Genbank to determine the two key epitopes of HBV PreS1 genotype (amino acid epitope 21-47 and 94-117, abbreviated as P21 and P94) and PreS1 full-length genotypes. The correlation among three genotyping approaches was analyzed by Cohen's kappa coefficient to verify the consistency between the key-epitope genotyping and the full-length preS1 genotyping. Results: 232 samples were successfully sequenced. The genotyping based on the kind of P21 epitope protein sequence, 201 cases for genotype C, 23 cases for genotype B and 8 cases for uncertain genotypes and genotyping based on the form of P94 epitope protein sequence, 199 cases for genotype C, 25 cases for genotype B and 8 cases for indeterminate genotypes. Lastly, the genotyping based on sequence of the full-length PreS1 sequence, 207 and 25 cases for genotype C and B. P21 or P94 epitope genotyping and PreS1 full length genotyping were highly consistent, respectively, 96.55% and 96.12%, and the two epitopes (P21and P94) genotyping have parallel consistency (93.10%). Conclusion: In this study, an innovatively genotyping method based on the amino acid sequence of key epitopes was proposed. The genotypes of HBV in china were mainly B and C genotypes, and the genotypes of key conserved epitopes of HBV PreS1 were highly consistent with the full-length genotyping ( > 96%). Moreover, genotyping with one or two key epitopes can be used in place of the full-length genotyping.
Assuntos
Vírus da Hepatite B , Sequência de Aminoácidos , China , Epitopos , Genótipo , Hepatite B , Anticorpos Anti-Hepatite B , Antígenos de Superfície da Hepatite B , Humanos , Reação em Cadeia da PolimeraseRESUMO
OBJECTIVE: In humans, stem cell factor (SCF), produced by cumulus granulosa cells during the follicular phase, plays a crucial role in follicular development. Remarkably, polycystic ovary syndrome (PCOS), one of the main reasons affecting women fertility, is accompanied by some abnormal follicles. Is there a relationship between SCF and PCOS? This study aimed to compare the expression of SCF in follicle and serum from patients with and without PCOS undergoing in vitro fertilization (IVF) treatment and to investigate the potential relationship between aberrant SCF expression and PCOS. PATIENTS AND METHODS: Serum, follicular fluid (FF) samples and granulosa cells (GCs) from 48 patients with PCOS (PCOS group) and 62 normal ovulatory patients (control group) were collected. SCF was evaluated in FF, serum, and GCs by using enzyme-linked immunosorbent assay, immunofluorescence staining, Western blot and real-time PCR. The rates of metaphase II (MII) oocyte, fertilization, embryo cleavage and high-quality embryo between PCOS group and control group were also analyzed. RESULTS: The rates of MII oocyte and fertilization were significantly lower in PCOS group than those in control group (p < 0.05). No difference was observed for the rate of embryo cleavage and high-quality embryo in these two groups. The concentrations of SCF in serum and FF from PCOS patients were remarkably lower than those in the controls (p < 0.05). Moreover, the expressions of SCF protein and SCF mRNA in GCs from PCOS patients were also decreased compared with the controls (p < 0.05). CONCLUSIONS: PCOS patients showed a reduced SCF expression in serum and follicle, which might be associated with oocyte dysmaturity and low fertilization rate.
Assuntos
Síndrome do Ovário Policístico/metabolismo , Fator de Células-Tronco , Feminino , Fertilização in vitro , Líquido Folicular , Células da Granulosa , Humanos , OócitosRESUMO
Purified recombinant mouse endostatin protein has been reported to regress established murine solid tumors by inhibiting the proliferation of endothelial cells. To develop a clinical gene therapy strategy with endostatin, we cloned the cDNA of human endostatin by RT-PCR from human placenta. A 150-bp sequence encoding the IgG leader peptide was fused in frame to the 5' end of the endostatin cDNA and recombinant adenoviruses, AdENDO-YFP and AdENDO, carrying endostatin gene expression cassettes were rescued. AdENDO-YFP infects cultured mammalian cells at high efficiency and expresses a biologically active human endostatin in secreted form at high levels both in vitro and in vivo. When delivered in vivo, a strain-specific expression pattern was observed, with the highest and longest endostatin expression in 129/J mice. After systemic delivery of 2 x 10(9) PFU of AdENDO-YFP into 129/J mice, human endostatin expression was achieved at a mean value of 1.34 +/- 0.42 microg/ml of serum (n = 6) and inhibition of lung metastasis was observed in an EOMA tumor model. However, high dose intravenous delivery of AdENDO-YFP and AdENDO was associated with severe acute toxicity in recipient mice that included loss of weight, bleeding, and death of animals. These events were not observed with the injection of identical doses of a control adenovirus that did not contain the endostatin gene. Because the endostatin adenovirus-associated acute toxicity was also observed in immunodeficient NCRNU-M nude mice, the toxicity does not appear to be the result of the immunogenicity against human endostatin or the EYFP protein.
Assuntos
Adenoviridae/genética , Inibidores da Angiogênese/genética , Colágeno/genética , Terapia Genética/métodos , Neoplasias Pulmonares/terapia , Melanoma Experimental/terapia , Fragmentos de Peptídeos/genética , Sequência de Aminoácidos , Inibidores da Angiogênese/metabolismo , Inibidores da Angiogênese/toxicidade , Animais , Antineoplásicos/metabolismo , Antineoplásicos/toxicidade , Sequência de Bases , Divisão Celular , Colágeno/metabolismo , Colágeno/toxicidade , Endostatinas , Endotélio Vascular/efeitos dos fármacos , Técnicas de Transferência de Genes , Humanos , Neoplasias Pulmonares/irrigação sanguínea , Neoplasias Pulmonares/patologia , Melanoma Experimental/irrigação sanguínea , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Nus , Dados de Sequência Molecular , Neovascularização Patológica/prevenção & controle , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/toxicidade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de AminoácidosRESUMO
Synergy between interleukin-12 (IL-12) and B7-1 (CD80) for cancer immunotherapy has previously been demonstrated in animal models of breast cancer, lymphoma, and multiple myeloma. With a view to human clinical application, tricistronic retroviral and adenovirus vectors co-expressing IL-12 (IL-12p40 plus IL-12p35) and CD80 were constructed by utilizing two internal ribosome entry site (IRES) sequences to link the three cDNAs. A murine stem cell virus (MSCV)-based retroviral vector (MSCV-hIL12.B7) utilized distinct IRES sequences from the encephalomyocarditis virus (EMCV) and the foot-and-mouth disease virus (FMCV), whereas Ad5-based adenovirus vectors contained transcriptional units with two EMCV IRES sequences under the control of murine (AdMh12.B7) or human (AdHh12.B7) cytomegalovirus promoters. AdMh12.B7 was found to consistently direct higher levels of IL-12 and CD80 expression than AdHh12.B7 following infection of a number of human tumor cell lines. In preclinical studies, the human myeloma cell line U266 was infected with MSCV-hIL12.B7 and a resulting clonal cell line, U/MSCV-h12.B7, was generated with stable expression of CD80 and secreting IL-12 at 1 ng/24 h/10(6) cells. By comparison, following AdMh12.B7 infection, 81% of infected U266 cells (U/AdMh12.B7) expressed CD80 and secreted IL-12 at 25-50 ng/24 h/10(6) cells. Both engineered myeloma cell lines stimulated enhanced allogeneic mixed lymphocyte proliferation and provoked increases in cytotoxic T-lymphocyte responses and gamma-interferon release from normal donor lymphocytes exposed to parental U266 cells. These results suggest potential clinical utility of AdMh12.B7 in immunotherapy strategies for the treatment of multiple myeloma and other cancers.
Assuntos
Adenoviridae/genética , Antígeno B7-1/genética , Vetores Genéticos , Imunoterapia/métodos , Interleucina-12/genética , Neoplasias/terapia , Retroviridae/genética , Células Tumorais Cultivadas/efeitos dos fármacos , Antígeno B7-1/metabolismo , Testes Imunológicos de Citotoxicidade , Citotoxicidade Imunológica , DNA Complementar , Quimioterapia Combinada , Citometria de Fluxo , Técnicas de Transferência de Genes , Humanos , Interleucina-12/metabolismo , Neoplasias/metabolismo , Neoplasias/virologia , Linfócitos T/imunologia , Células Tumorais Cultivadas/metabolismo , Células Tumorais Cultivadas/virologiaRESUMO
Interleukin-6 (IL-6) is implicated in the in vivo proliferation of malignant plasma cells in multiple myeloma. To define the molecular basis of the IL-6-induced mitogenic response in myeloma cells, we applied STAR (subtractive transcriptional amplification of mRNA), a new differential expression analysis technology, to isolate mRNAs preferentially expressed in IL-6-treated versus untreated cultures of the factor-responsive myeloma cell line U266. From the resulting collection of STAR clones, sequence information was obtained for a total of 72 distinct transcripts. Of these, 29 were found to correspond to known genes, 22 matched expressed sequence tags in public databases and 21 showed no sequence similarity to any existing entries. Among the known genes uncovered in the screen were those encoding proteins that function in cell division, cell signalling and gene/protein expression. Northern blot analysis documented that two transcription factor genes chosen for further study, c-myc promoter-binding protein (MBP-1) and X-box binding protein 1 (XBP-1), were up-regulated in U266 cells about 3-fold relative to the cell cycle-dependent beta-actin gene 12 h after IL-6 treatment. Both genes were also similarly up-regulated by IL-6 in factor-dependent ANBL-6 myeloma cells. These results indicate that MBP-1 and XBP-1 are IL-6 genes in myeloma cells; as such, they may play a role in IL-6-mediated growth control in multiple myeloma.
Assuntos
Proteínas de Ligação a DNA/genética , Interleucina-6/farmacologia , Mieloma Múltiplo/genética , Proteínas de Neoplasias/genética , Fosfopiruvato Hidratase , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor , Sequência de Bases , Biomarcadores Tumorais , Divisão Celular/efeitos dos fármacos , DNA Complementar/genética , Etiquetas de Sequências Expressas , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-6/fisiologia , Dados de Sequência Molecular , Mieloma Múltiplo/patologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/isolamento & purificação , RNA Neoplásico/biossíntese , RNA Neoplásico/genética , RNA Neoplásico/isolamento & purificação , Fatores de Transcrição de Fator Regulador X , Transdução de Sinais/efeitos dos fármacos , Técnica de Subtração , Transfecção , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/metabolismo , Proteína 1 de Ligação a X-BoxRESUMO
Increasing evidence suggests a role for immunologic vaccination and therapy in the management of minimal residual myeloma. We have previously demonstrated a synergistic effect of combining the Th1 stimulating cytokine IL-12 with the co-stimulatory molecule CD80 in murine myeloma vaccination therapy. We reasoned that the efficacy of such treatment might be further improved by incorporating additional gene products which enhance the function of antigen presenting cells. Studies were therefore conducted with murine myeloma BM1 cells expressing Flt3L (membrane bound or soluble forms) or GM-CSF and the IL-12 x CD80 combination. Single agent and combined therapeutic approaches were explored. All gene-modified BM1 cells, except BM1/IL-12 x CD80, developed tumors when subcutaneously injected into BALB/c mice. As prophylactic tumor vaccines, the combined use of gene-modified BM1/sFlt3L+GM-CSF+IL-12 x CD80 was most effective, providing 100% protection against subsequent parental BM1 tumor challenge. By comparison, only partial protection was observed with any single gene-engineered tumor vaccine. Notably, IL-12 x CD80 coexpressing BM1 cell vaccines were the most effective therapeutic vaccine in a minimal disease model. Such protective vaccination was achieved by stimulation of lymphocyte proliferation and enhancement of cytotoxic lymphocyte activity.
Assuntos
Vacinas Anticâncer/uso terapêutico , Terapia Genética , Imunoterapia , Mieloma Múltiplo/prevenção & controle , Adjuvantes Imunológicos/metabolismo , Adjuvantes Imunológicos/uso terapêutico , Animais , Antígeno B7-1/metabolismo , Antígeno B7-1/uso terapêutico , Vacinas Anticâncer/metabolismo , Divisão Celular , Terapia Combinada , Citotoxicidade Imunológica , Feminino , Técnicas de Transferência de Genes , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/uso terapêutico , Humanos , Interleucina-12/metabolismo , Interleucina-12/uso terapêutico , Ligantes , Proteínas de Membrana/metabolismo , Proteínas de Membrana/uso terapêutico , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/terapia , Retroviridae/genética , Baço/imunologia , Taxa de Sobrevida , Linfócitos T/imunologia , Linfócitos T/metabolismo , Linfócitos T Citotóxicos/imunologia , Células Tumorais Cultivadas , VacinaçãoRESUMO
We constructed a plasmid pBMTHBR2 containing mouse MT promoter, entire HBsAg gene (preS and S gene), splicing signal from SV40 and complete BPV genome. Using calcium phosphate precipitation technique to transform C127 Cells and using BPV transforming foci as selective markers, we obtained transformed cell clones. The experiment shows that 57% of the cloned cell lines can secrete HBsAg continuously.
Assuntos
Papillomavirus Bovino 1/genética , Antígenos de Superfície da Hepatite B/metabolismo , Vírus da Hepatite B/genética , Papillomaviridae/genética , Plasmídeos/genética , Animais , Sequência de Bases , Transformação Celular Viral/genética , Células Clonais , Regulação Viral da Expressão Gênica , Camundongos , Regiões Promotoras GenéticasRESUMO
We have previously cloned two mouse homeobox genes Tlx-1 (T-cell leukemia homeobox gene-1) and a related gene Tlx-2 based on their homology to human HOX11, a putative proto-oncogene involved in human T-cell leukemia. We have mapped Tlx-1 to mouse chromosome 19 and Tlx-2 to chromosome 6 by linkage analysis using an interspecific backcross (C57BL/6J x Mus spretus) F1 x M. spretus. The proposed gene orders and genetic distances for Tlx-1 and Tlx-2 are centromere 19-Lpc-1-(25.53 cM)-Pltr-4-(5.32 cM)-Tlx-1- (3.19 cM)-Ins-1-(7.45 cM)-Xmv-18, and centromere 6-Tcrb-(12.90 cM)-Mltr-3-(10.75 cM)-Tlx-2-(18.42 cM)-Xmv-6.
Assuntos
Mapeamento Cromossômico , Genes Homeobox , Proteínas de Homeodomínio , Leucemia de Células T/genética , Animais , Proteínas de Ligação a DNA/genética , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Oncogênicas/genética , Proto-Oncogene Mas , Proteínas Proto-OncogênicasRESUMO
Recent epidemiological studies have identified plasma triglyceride as a risk factor for atherogenesis. We have generated a mouse transgenic line that carries a recessive mutation designated lpd (lipid defect). Homozygous lpd mice develop as runts and die by age 10-15 days with striking liver pathology characterized by the presence of numerous lipid-containing vacuoles and extensive accumulation of triglycerides. Cloning of the mutant insertion locus and the wild-type lpd locus have revealed a duplication of host genomic sequences at the site of integration. Mapping of the lpd locus with the Jackson Laboratory BSS interspecific backcross panel of (C57BL/6JEi x SPRET/Ei) F1 x SPRET/Ei placed the lpd locus to the distal part of chromosome 16. These observations suggest that the transgene disrupts a putative gene at the lpd locus and that lpd is a novel locus related to triglyceride metabolism. The lpd mutant mice may serve as models for human disorders of fatty livers or hypertriglyceridemia.
Assuntos
Triglicerídeos/metabolismo , Animais , Southern Blotting , Colesterol/sangue , Mapeamento Cromossômico , Clonagem Molecular , Modelos Animais de Doenças , Fígado/metabolismo , Fígado/patologia , Camundongos , Camundongos Transgênicos , Família Multigênica , Mutagênese , MutaçãoRESUMO
We have previously generated a mouse transgenic line with an insertional mutation designated lpd that demonstrates a phenotype of hypertriglyceridemia and fatty liver. Since the recently identified phosphatidylserine-specific phospholipase A1 (PS-PLA1) demonstrates significant homology to triglyceride lipases, we reasoned that the mouse Ps-plaI gene may be the disrupted gene within the lpd locus. Using a rat PS-PLA1 cDNA sequence to search the EST database, we identified a mouse EST homolog AA839424. Sequencing analysis of AA839424 revealed a putative Ps-pla1 protein of 456 amino acids with extensive overall structural conservation with human and rat PS-PLA1 and with triglyceride lipases. Conserved sequences in Ps-pla1 include a lipase consensus sequences GxSxG, a catalytic triad, and eight of the ten conserved cysteine residues that are required for tertiary structure. Mouse Ps-plal carries a phosphatidylserine-binding motif that is absent in all triglyceride lipases. Using a mouse whole-genome radiation hybrid (WG-RH) mapping panel (T31), we mapped mouse Ps-pla1 to Chromosome (Chr) 16 between genetic markers D16Mit194 and D16Mit38, which is 17.1 cM centromeric to the lpd locus. On the basis of chromosome location, we conclude that Ps-pla1 and lpd are distinct genes in lipid metabolism.