ABSTRACT
Objective To screen for malignant tumors and high-risk factors in rural residents over 60 years old， so as to prevent and control the occurrence and development of tumors in the future. Methods The survey was conducted with reference to part of the questionnaire in the "Urban Cancer Early Diagnosis and Treatment Project and Evaluation of High-risk Populations". Clinical examinations included serum tumor marker detection， CT screening for lung cancer， occult blood （+） plus colonoscopy screening for colorectal cancer， and mammography screening. Individuals who were positive in the abovementioned clinical tests were defined as high-risk subjects. Results A total of 271 high-risk subjects （1.91%） were screened out of 14 161. Among the high-risk subjects， 71 cases of malignant tumors （26.19%） were found， with an incidence rate of 501.38 per 105. The top five tumors （63.38% of all diagnosed） were mainly concentrated in lung， upper digestive tract， blood system， urinary system， and rectum-colon. The proportion of malignant tumors detected by positive indicators was 61.54% of blood； 46.15% of carcinoembryonic antigen and carbohydrate antigen 125； 23.08% of alpha-fetoprotein； 16.66% of lung CT； and 3.09% of prostate PSA. The positive indicators in the high-risk subjects were mainly for the tumors in the prostate， lungs， liver， and CEA/CA125. The subjects with positive test indicators had lower average annual income in the last 5 years than the normal subject group （χ2=3.380， P=0.040）. The subjects with positive test indicators had higher proportion in family history of tumors than the normal group （χ2=2.596， P=0.046）. People in thehigh-risk group had a higher proportion than the normal group in suffering from hypertension， liver disease， gastrointestinal disease， respiratory system disease， and surgical treatment. Patients with high-risk tumors were found to have higher proportion than the normal group in showing pre-tumor clinical symptoms in the last 1 year. Study of the tumor-related risk factors found that the high-risk group had a higher proportion of high-fat/high-cholesterol diet， alcohol drinking， passive smoking， and personality depression. Conclusion High tumor risk factors have been identified in this population. It is necessary to strengthen the corresponding intervention and follow-up treatment of precancerous diseases in the future. We recommend the government to conduct tumor screening among high-risk groups to improve cost-effectiveness.
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Objective To compare the efficacy of two doses of ambroxol in the treatment of the elderly patients with acute exacerbation of chronic obstructive pulmonary disease (COPD).Methods A hundred and ten cases of the elderly patients with an acute exacerbation of COPD were divided into low-dose group and high-dose group.Low-dose group (56 cases) were given Ambroxol 30 mg,2 times per day for 10 day and high-dose group (54 cases) were given Ambroxol 60 mg,2 times per day for 10 days.Results The effective rate in high-dose group was significantly higher than that in low-dose group.Conclusion High-dose ambroxol has significant effect in the treatment of acute exacerbation of chronic obstructive pulmonary disease.
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Objective To investigate the CD34~+ umbilical cord blood hematopoietic stem cells (CD34~+ UBSC) transfected with interleukin 21 (IL-21) against the ovarian cancer effect in tumor-bearing nude mice. Methods CD34~+ UBSC were obtained from the UBSC by a magnetically activated cell sorting technique and then transfected with recombinant plasmid plRES2-IL-21-EGFP after the CD34~+ UBSC were proliferated in vitro. The therapeutic effect was evaluated by the size of the tumor and the life span in nude mice treated with the CD34~+ UBSC-IL-21. The expression of IL-21 and its bioactivity in CD34~+ UBSC-IL-21 and in local neoplasitc tissues were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR), immune fluorescence technique, ELISA, Western blot, immunohistochemistry and splenocyte proliferative activity. The NK cell cytotoxicity and the numbers of NK cells, serum level of IFN-γ and TNF-αwere simultaneouly detected by FCM and ELISA, respectively. Results CD34~+ UBSC were cultured and transfected with pIRES2-IL-21-EGFP successfully. CD34~+ UBSC-IL-21 could inhibit the tumor growth and extended nude mice life span compared with other groups (P < 0.01). The expression of IL-21 in the neo-plastic tissue, serum level of IFN-γ and TNF-α , NK cell activity and the numbers of NK cells of mice origin and of human origin in splenocytes were increased significantly in the nude mice treated with CD34~+ UBSC-IL-21 compared with other groups(P <0.01). Conclusion The CD34~+ UBSC transfected with IL-21 have competent against ovarian cancer in tumor-bearing nude mice. The findings may establish a foundation for gene therapy of the ovarian cancer by CD34~+ UBSC-IL-21 in clinic application.
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Sox9 gene was cloned from immortalized precartilaginous stem cells and its eukaryotic expression vector constructed in order to explore the possibility of bone marrow-derived stromal cells differentiation into precartilaginous stem cells induced by Sox9. A full-length fragment of Sox9 was obtained by RT-PCR, inserted into pGEM-T Easy clone vector, and ligated with pEGFP-IRES2 expression vector by double digestion after sequencing. The compound plasmid was transfected into born marrow-derived stromal cells by Lipofectamine 2000, and the transfection efficacy and the expression of Sox9 and FGFR-3 were observed. Flow cytometry was used to identify the cell phenotype, and MTT was employed to assay proliferative viability of cells. Sequencing, restrictive endonuclease identification and RT-PCR confirmed that the expansion of Sox9 and construction of Sox9 expression vector were successful. After transfection of the recombinant vector into bone marrow-derived stromal cells, the expression of Sox9 and FGFR-3 was detected, and proliferative viability was not different from that of precartilaginous stem cells. It was concluded that Sox9 gene eukaryotic expression vector was successfully constructed, and the transfected bone marrow-derived stromal cells differentiated into the precartilaginous stem cells.
Subject(s)
Base Sequence , Bone Marrow Cells/cytology , Cartilage/cytology , Cell Differentiation/genetics , Cells, Cultured , Cloning, Molecular , Genetic Vectors/genetics , Molecular Sequence Data , Receptor, Fibroblast Growth Factor, Type 3/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , SOX9 Transcription Factor/biosynthesis , SOX9 Transcription Factor/genetics , Stem Cells/cytology , Stromal Cells/cytology , TransfectionABSTRACT
BACKGROUND: Precartilaginous stem cells (PSCs) have strong proliferation ability and differentiation potential,but they are instable and prone to differentiate.Importing exogenous gene could immortalize them and leave phenotype character unchanged.OBJECTIVE: To establish immortalized precartilaginous stem cells (PSCs) from neonatal SD rats in vitro for the further related research about the differentiation mechanism and clinical application of precartilaginous stem cells.DESIGN,TIME AND SETTING: Single sample observation.The study was carried out in the Department of Orthopedics.Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from October 2005 to September 2006.MATERIALS: Neonatal SD rats,irrespective of gender,24-hour old,were used for prepare PSCs.METHODS: By using LipofectamineTM 2000,a gene transfection reagent,plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigene gene (SV40Tag) was transfected into the primary cultured PSCs isolated by immuniomagnetic beads coasted with the second antibody.Colonies were isolated by puromycin selection and expanded by many passages.MAIN OUTCOME MEASURES: Biological character of PSCs; plasmid identification; biological character of transfected cells and identification; RT-PCR; growth curve.RESULTS: Immunomagnetic beads separation system obtains PSCs,which was confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive PSCs.Double restriction enzyme was cut,electrophoresis confirmed pCMV was 3 kb,SV40T was 2.3 kb.A particular anti-puromycin cell clone was acquired,which was confirmed as FGFR-3 positive PSCs.The total RNA was isolated from the positive cell clones,and a 588 bp fragment,which was specific for the SV40T antigene gene,was amplified.The transfected cells were expanded to immortalized cell strain,named as immortalized precartilaginous stem cells (IPSCs).Thepopulation doubling time of IPSCs was (22.98±2.77) hours,no significant effect of subculture,freezing and recovering had been found.CONCLUSION: Precartilaginous stem cells could be isolated from neonatal SD rats,cultured in vitro,and immortalized through the transfection of pCMVSV40T/PUR.
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Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.
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Objective To observe the effect of Huangqi (Radix Astragali seu Hedysari) Injectio on the abnormal expression of aquaporin-1 (AQP-1) caused by glucose lactate peritoneal dialysis solution. Methods Different glucose concentration lactate peritoneal dialysis solutions and Huangqi Injectio given to the rats intraperitoneally. Eight weeks later, the ultrafiltration of peritoneal dialysis, peritoneum clearance, peritoneal structure, AQP-1 and its gene expression were observed. Results Compared with the control group, in the glucose and Huangqi groups, the ultrafiltration of peritoneal dialysis was remarkably decreased (P