This paper outlines some molecular marking technology based on rDNA sequences and several DNA fingerprinting technology (RAPD, ARDRA, AFLP, REP/ERIC-PCR) used in classification, identifica-tion and diversity research in lactic acid bacteria. The principles, methods and progress in recent years of these technologies were also introduced. At the same time, this paper also compares the advantages and dis-advantages of these methods. People should choose suitable method according to their purposes.

Objective To investigate the efficacy,side effects and cosmetic effects of breast-conserving operation combined with radiotherapy for patients with breast cancer. Methods Forty-three patients with early stage of breast cancer(n=30 for stage I and n=13 for stage II) were performed extended resection plus axillary lymph node dissection or quadrant resection plus axillary lymph node dissection.After operation,tangential field irradiation was conducted with 50 Gy of X ray,tumor bed was irradiated with additional 10 Gy of electron beam,and supraclavicular field irradiation was also administered with 30 Gy of X ray and 22 Gy of electron beam. Results There was no recurrence during the follow up for 14 to 50 months.The acute side effects of radiotherapy included breast complaints(n=10,grade Ⅰ,23.26%),skin erythema and hyperpigmentation(n=19,grade Ⅰ,44.19%),moist desquamation(n=5,grade Ⅱ,11.63%) and radiation esophagitis(n=4,grade Ⅰ,9.30%),while one patient experienced radiation esophagitis one month after radiotherapy.No such complications as skin fibrosis and necrosis,angiotelectasis,breast and upper extremity edema occurred.The short-term cosmetic effects were satisfactory to 93.02% of all the patients. Conclusion For early stage of breast cancer(stage I and stage Ⅱ),breast-conserving operation combined with radiotherapy may yield satisfactory survival rate and short-term cosmetic effects,as well as low incidences of side effects.

Objective To evaluate the clinical features,therapies and prognosis in patients with mucosa-assoeiated lymphoid tissue (MALT) lymphoma in ocular adnexal marginal zone (OAML).Methods A retrospective analysis was made upon clinical data from 21 patients with OAML admitted into Beijing Tongren Hospital from June,2008 to December,2011.Results There were 12 (57.1 ％) men and 9(42.9％) women,with a median age of 57 (23-79) years old.Majority of patients had localized pathological changes.Among them,16 patients (76.2％) were in stage Ⅰ E,and 5 (23.8％) in stage Ⅳ E.Surgical resection as the sole treatment was performed in 13 patients (61.9％),and positron emission tomography CT(PET-CT) imaging demonstrated normal fluorine 18-fluorodeoxyglucose (FDG) uptake after surgical resection,who were managed with no further therapy.All the 13 patients were followed up for median 14 (5-38) months,and all in complete remission.Combination chemotherapy was given to 8(38.1％) patients.Three patients in stage Ⅰ E treated with COP (cyclophosphamide,vineristine and prednisone) or CHOP (cyclophosphamide,adriamycin,vincristine and prednisone) were all in partial remission.Five patients in stage ⅣE were treated with COP/CHOP in combination with rituximab,and all in complete remission.The 3-year overall survival rate and disease-free survival rate in the total patients were 100.0％ and 74.9％ respectively.Conclusions The patients with OAML generally have localized disease,show indolent clinical course,and present low lymphoma-related mortality.Surgical resection is a very important treatment in the patients with local disease.Systemic chemotherapy should be considered in patients at advanced stages.Rituximab in combination with chemotherapy can improve the remission rate.

The aim of this study was to analyze the efficacy of first-line chemotherapy in treating patients with ocular adnexal extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT). Eight consecutive newly diagnosed ocular adnexal MALT lymphoma patients were treated with chemotherapy, in which 3 patients in stage IE were treated with a combination of cyclophosphamide, vincristine and prednisolone (COP) or cyclophosphamide, adriamycin, vincristine, and prednisolone (CHOP), 5 patients in stage IV were treated with COP/CHOP in combination with rituximab. The results showed that chemotherapy resulted in an overall response rate of 100%, 3 patients in stage IE were in partial remission (PR), 5 patients in stage IV were in complete remission (CR). After a median follow-up of 21 months, 3 patients in stage IE were still in PR status, 5 patients in stage IV were still in CR status, and no relapses or disease progression were observed. It is concluded that the first-line chemotherapy has been confirmed to be effective and well tolerated in patients with localized ocular adnexal MALT lymphoma. Rituximab combined with chemotherapy can increase remission rate, it is feasible option as first-line treatment for ocular adnexal MALT lymphoma.
Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Eye Neoplasms ; drug therapy ; Female ; Humans ; Lymphoma, B-Cell, Marginal Zone ; drug therapy ; Male ; Middle Aged

This study was to investigate the anti-lymphocytic malignancy immunologic effects induced by two types of the idiotypic nucleic acid vaccines which were constructed from the genomic DNA and RNA of the human B lymphoma cell line respectively. Namalwa cell line and BALB/c mice were used as the models. The gene fragments of the IgH variable region (IgHV), which were obtained from the genomic DNA and RNA of Namalwa cell respectively, were cloned into the eukaryocytic expression vector pcDNA 3.0 to be used as the idiotypic nucleic acid vaccines. After transfecting COS cells with one of vaccines constructed from the genomic DNA by using LipofectAMINE, the result of transcription was identified by using RT-PCR. The experimental mice were immunized by intramuscular injection with two types of vaccines. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The results showed that the nucleic acid vaccine constructed from the genomic DNA can be transcribed in COS cells, the transcription product turned shorter, and the intron region of 86 bp was spliced accurately. When immunizing the mice, two vaccines both induced the anti-idiotypic antibody against Namalwa cell, the anti-idiotypic antibody could be detected since detected since after immunization, and got to the peak of titer on the sixth week. It was concluded that the nucleic acid vaccines against lymphoma can be constructed from both the genomic DNA and RNA.
Animals ; Antibodies, Anti-Idiotypic ; blood ; immunology ; Base Sequence ; COS Cells ; DNA, Neoplasm ; genetics ; immunology ; Humans ; Immunoglobulin Heavy Chains ; genetics ; immunology ; Immunoglobulin Variable Region ; genetics ; immunology ; Injections, Intramuscular ; Lymphoma, B-Cell ; genetics ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Plasmids ; administration & dosage ; genetics ; immunology ; Time Factors ; Tumor Cells, Cultured ; Vaccines, DNA ; genetics ; immunology

OBJECTIVETo analyze the phenotypic and functional characteristics of endothelial (T3A) cells derived from human hepatocellular cell carcinoma.

METHODSEndothelial cells were isolated from human hepatocellular carcinoma specimens. The identification of T3A cells was performed by checking von Willebrand Factor (vWF), CD31, CD34 and Dil-Ac-LDL uptake. The cell surface fenestrations, a specific morphological feature of tumor derived EC, were investigated by scanning and transmission electron microscopy. The phenotypic characteristics of T3A cells were analyzed by fluorescence-activated cell sorter (FACS) and were further conformed by real-time PCR at transcription level. Furthermore, tumor necrosis factor alpha (TNFalpha)-induced cytotoxicity was evaluated by 3-(4, 5-dimethythiazolyl) -2, -diphenyl-2H-tetrazolium-bromide (MTT) assay; Matrix metalloproteinase secretion was detected by zymography; Angiogenic ability in vitro was analyzed by culturing T3A cells in three-dimensional Matrigel plug. Coagulant and fibrinolytic activities were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe isolated T3A cells exhibited classic "spindle-shape" morphology and monolayer growth and contact inhibition properties. Immunofluorescent staining showed that T3A cells expressed vWF, CD31, CD34, and uptake of Dil-Ac-LDL at a high level. The cell surface fenestrations were observed on T3A cells by scanning and transmission electron microscopy. By FACS and real-time PCR, T3A cells were found to express alphav3, alphavbeta5 and TNF receptor p75 at high levels, and TNF receptor p55 and ICAM-1 at low levels, as compared with those in human liver sinusoidal endothelial cells (LSEC). In response to TNFalpha, LSEC exhibited a dose-dependent cytotoxicity, while T3A cells were resistant. Gelatin zymography showed that MMP-2 activity was higher in T3A cells than that in LSEC. In a three-dimensional plug of Matrigel, T3A cells exhibited stronger angiogenic ability as compared with LSEC. In addition, T3A cells released more tissue factor (TF), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1) and urine plasminogen activator (u-PA) than LSEC in response to TNFalpha.

CONCLUSIONTumor-derived endothelial cells are phenotypically and functionally different from those derived from normal liver tissue.

Antigens, CD34 ; metabolism ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Cell Shape ; Cells, Cultured ; Endothelial Cells ; metabolism ; pathology ; ultrastructure ; Gene Expression ; Humans ; Integrin alphaVbeta3 ; metabolism ; Integrins ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Lung ; blood supply ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; metabolism ; Microscopy, Electron, Scanning ; Neovascularization, Pathologic ; metabolism ; pathology ; Phenotype ; Plasminogen Activator Inhibitor 1 ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Tumor Necrosis Factor, Type I ; metabolism ; Receptors, Vitronectin ; metabolism ; Tissue Plasminogen Activator ; metabolism ; Tumor Cells, Cultured ; Tumor Necrosis Factor Decoy Receptors ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; von Willebrand Factor ; metabolism

OBJECTIVETo explore the feasibility and safety of enteral nutrition in preoperative bowel preparation for rectal cancer patients undergoing radical operation.

METHODSSixty rectal cancer patients undergoing selective low anterior resection were randomized into the trial group(n=30) and the control group(n=30). Patients in the trial group received clean liquid integral protein diet for 3 days before operation without mechanical bowel preparation. Patients in the control group received traditional diet and mechanical bowel preparation. The intraoperative and postoperative clinical data, the quality of bowel preparation, postoperative complications, and nutritional parameters were compared between the two groups.

RESULTSThere were no significant differences in clinicopathological characteristics between the two groups before operation. The operative time, blood loss, quality of bowel preparation as well as postoperative hospital stay were not significantly different(all P>0.05). While the time to first flatus [(2.53±0.91) d vs. (3.03±0.68) d] and semi-liquid diet intake[(3.95±0.83) d vs. (4.52±1.14) d] were significantly shorter in the trial group as compared with the control group(all P<0.05). There were no death and no significant difference in postoperative complications [16.7%(5/30) vs. 20.0%(6/30), P>0.05]. The levels of postoperative total protein, albumin, and prealbumin decreased significantly. Meanwhile, the levels of postoperative albumin[(36.2±2.5) g/L vs. (33.5±2.6) g/L, P<0.01] and prealbumin [(325.4±28.2) mg/L vs. (302.5±34.2) mg/L, P<0.01] in the trial group were significantly higher than those in the control group.

CONCLUSIONSPreoperative enteral nutrition can replace the mechanical bowel preparation with better efficacy, and improve the postoperative nutritional status without increasing surgical risk in rectal cancer patients undergoing radical operation.

Digestive System Surgical Procedures ; adverse effects ; Enteral Nutrition ; Humans ; Postoperative Complications ; Preoperative Care ; methods ; Rectal Neoplasms ; surgery

OBJECTIVETo investigate whether S. aureus could activate NF-κB signaling pathway in human osteoblasts.

METHODSImmunoblot and electrophoretic mobility shift assay were used to detect the degradation of I-κBα and activation of NF-κB in human osteoblasts following infection with S.aureus, respectively, and there were investigated the activated state of NF-κB signaling pathway in human osteoblasts. In addition, enzyme-linked immunosorbent assay was used to measure the secretion of IL-6 in culture supernatants, which was represented as one of important cytokines in osteomyelitis, and an inhibitor of NF-κB, SN50, which was added to human osteoblasts culture prior to 1 hour at 50 µmol/L before the infection of S.aureus, was used to determine whether S.aureus-activated NF-κB signaling pathway regulates IL-6 secretion of human osteoblasts.

RESULTSS.aureus could induce the degradation of I-κBα (I-κBα(15 min)/I-κBα(0 min) = 0.409 ± 0.245 and I-κBα(30 min)/I-κBα(0 min) = 0.061 ± 0.010) and activation of NF-κB in human osteoblasts in a time and dose-dependent manner following infection. In addition, the secretion of IL-6 in the supernatants of human osteoblasts ((2.17 ± 0.11) µg/L) was suppressed by 50 µmol/L SN50 compared to without the addition of SN50 ((3.58 ± 0.31) µg/L) (F = 174.25, P < 0.05).

CONCLUSIONSS.aureus could activate NF-κB signaling pathway in human osteoblasts, which could regulate cytokines secretions of human osteoblasts.

Cells, Cultured ; Humans ; Interleukin-6 ; secretion ; NF-kappa B ; metabolism ; Osteoblasts ; metabolism ; Signal Transduction ; Staphylococcal Infections ; metabolism

OBJECTIVETo investigate the role of mutation and mRNA expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 (PTEN) in tumorigenesis and progression of oral squamous cell carcinoma (OSCC).

METHODSThe mutation of exon 3, 5, 6 and exon 8 of PTEN gene in 42 oral squamous cell carcinoma tissue and paired normal tissue were detected by polymerase chain reaction (PCR) and DNA sequencing methods. The levels of PTEN mRNA expression in these tissues were assayed using semi-quantitative RT-PCR. Data was analyzed with SPSS 14.0 software package.

RESULTSMutated exon 5 of PTEN gene was found in 2 cases of advanced OSCC. The expression of PTEN mRNA was detected in all OSCC and paired normal tissue. The level of PTEN mRNA in OSCC tissue (0.36 +/- 0.12) was significantly lower than that of paired normal tissue (0.64 +/- 0.09, P < 0.01).

CONCLUSIONSThe decreased expression of PTEN mRNA contributes to tumorigenesis of OSCC.

Carcinoma, Squamous Cell ; genetics ; Chromosomes, Human, Pair 10 ; Exons ; Gene Expression ; Genes, Tumor Suppressor ; Humans ; Microfilament Proteins ; Mouth Neoplasms ; genetics ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphoric Monoester Hydrolases ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; metabolism ; Tensins

OBJECTIVETo establish the characteristic mode of HPLC fingerprint of Fufangteng combination formula for the quality control of this medicine.

METHOD10 batch certified products were selected to establish the characteristic mode of HPLC fingerprint of Fufangteng combination formula by chromatographic pattern matching and retention time correction.

RESULTThis characteristic mode of HPLC fingerprint can clearly differentiate the eligible Fufangteng products from the unqualified products.

CONCLUSIONThis method can provide thorough information for the quality control of Fufangteng combination formula.

Administration, Oral ; Astragalus membranaceus ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Euonymus ; chemistry ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control