ABSTRACT
Objective:To evaluate the compatibility laws of effective-component compatibility of Bufei Yishen formula Ⅲ (ECC-BYFⅢ) in regulating mucus hypersecretion of chronic obstructive pulmonary disease (COPD).Methods:According to the efficacy of the original Chinese medicine, the components of ECC-BYFⅢ were divided into four categories: Buqi (Ginsenoside Rh1+Astragaloside), Bushen (Icariin), Huatan (Nobiletin), and Huoxue (Paeonol). The four categories were divided into 14 groups based on the method of mathematical permutation. ① The rats were divided into control group, model group, ECC-BYFⅢ, and different components compatibility groups according to the random number table, totaling 17 groups. COPD rat model in stable phase was established by cigarette smoke exposure combined with repeated bacterial infections. The corresponding drugs were given by gavage at the 9th week of modeling, and the samples were collected at the end of the 16th week. The levels of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors of metalloproteinase 1 (TIMP-1) in serum and bronchoalveolar lavage fluid (BALF), and the levels of mucin (MUC) 5AC in lung tissue and BALF were detected by enzyme linked immunosorbent assay (ELISA). ② Human lung epithelial cells BEAS-2B were divided into blank group, model group, and different components compatibility groups. Hypoxia-induced mucus hypersecretion model of human lung epithelial cells BEAS-2B was established 4 hours after corresponding drug pretreatment. The mRNA expressions of MUC5AC, MUC5B, and MUC1 were detected by quantitative polymerase chain reaction (PCR). The mucus secretion indexes of rats and BEAS-2B cells were evaluated by Region (R) value comprehensive evaluation method.Results:① Compared with the control group, MMP-9 in serum and BALF from the model group were significantly increased, the level of TIMP-1 was significantly decreased, and MUC5AC in lung tissue and BALF were significantly increased. The results of R value comprehensive evaluation showed that except for the Buqi and Bushen groups, ECC-BYFⅢ and other components compatibility groups significantly corrected mucus hypersecretion in COPD rats, ECC-BYFⅢ, Bushen Quxie, Fuzheng Huatan, and Quxie groups were much better (R values were 2.15±0.42, 2.11±0.23, 2.16±0.23 and 2.16±0.55, respectively), compared with the model group (R value: 3.00±0.00), the differences were statistically significant (all P < 0.05). ② Compared with the blank group, the mRNA expressions of MUC5AC, MUC5B, and MUC1 increased in the model group. But different components compatibility groups had no significant effects on the mucus secretion of BEAS-2B cells. ③ The comprehensive evaluation results of R value about each in vivo and in vitro index showed that ECC-BYFⅢ, Huoxue, Quxie, Bushen Huoxue, Fuzheng Huatan, Buqi Quxie groups significantly corrected the mucus hypersecretion (R values were 2.30±0.43, 2.33±0.44, 2.12±0.68, 2.27±0.64, 2.24±0.27 and 2.29±0.47, respectively), compared with the model group (R value: 3.00±0.00), the difference was statistically significant (all P < 0.01). The order was: Quxie > Fuzheng Huatan > Bushen Huoxue > Buqi Quxie > ECC-BYFⅢ > Huoxue. Conclusions:Different components compatibility of ECC-BYFⅢ had different effects on COPD mucus secretion. The components containing Huatan (Nobiletin) or Huoxue (Paeonol) showed a better inhibitory effect on mucus secretion.
ABSTRACT
Objective To study the value of TCT, human papillomavirus DNA combined with colposcopy in the early screening of cervical cancer. Methods The clinical data of 80 cases of patients with cervical lesions in our hospital from January 2011 to January 2017 were retrospectively analyzed. Results Among the 80 cases of patients, ASCUS was detected by TCT 8 cases, LSIL6 cases, HSIL4 cases, SCC2 cases, the detection rate of cervical lesions was 25.0% (20/80); Normal and inflammatory 60 cases, accounting for 75.0% (60/80) of the total. HPV-DNA positive 28 cases, negative 52 cases, the positive rate was 35.0% (28/80); The detection rate of TCT (+) HPV (+) was significantly higher than TCT (+) HPV (-), TCT (-) HPV (+), TCT (-) HPV (-) (P<0.05), the detection rate of SCC reached 100.0%; The detection rate of CIN I, CIN II, CIN III of TCT examination, HPV-DNA examination combined with colposcopy were significantly higher than TCT+HPV-DNA examination (P<0.05), but the difference of detection rates of SCC between the two was not significant. Conclusion The value of TCT, human papillomavirus DNA and colposcopy in the early screening of cervical cancer is higher, so is worthy of promotion and use in the clinical.
ABSTRACT
BACKGROUND:CTLA-4Ig as a tolerance-induction agent is a potential strategy in graft-versus-host disease prevention. OBJECTIVE:To investigate the efficacy of CTLA4Ig-gene-modified bone marrow stromal cels mediated by adenovirus to induce T-cel tolerance of haploidentical donors. METHODS: The bone marrow stromal cels isolated culture from the bone marrow of HLA haploidentical donors were transfected by recombinant adenovirus encoding CTLA4IgcDNA (AdCTLA4Ig) at a multiplicity of infection=50 for 72 hours. The expression rate and the location of CTLA4Ig in the transfected cels were detected by fluorescence microscope after immunofluorescence staining. CTLA4Ig-modified bone marrow stromal cels (2×104, 4×104and 8×104) were respectively co-cultured with 105 T cels from the peripheral blood of HLA haploidentical donors and 105 peripheral blood mononuclear cels from recipients. The proliferative inhibition rate was determined by MTT assay, and the level of interleukin-2 in the supernatant was detected by ELISA. The bone marrow mononuclear cels (1×105/wel) were co-cultured with CTLA4Ig-modified bone marrow stromal cel layers constructed in 6-wel plates. The number of bone marrow mononuclear cels and colony-forming unit-granulocyte macrophages were calculated after 5-day culture. RESULTS AND CONCLUSION: The expression rate of CTLA4Ig at the multiplicity of infection=50 was as high as 85%, and the immunofluorescence signals of CTLA4Ig were distributed unevenly in the cytoplasm. The inhibition rates of 2×104, 4×104, and 8×104 CTLA4Ig-modified bone marrow stromal cels on proliferation of T cels were higher than that of untransfected cels. The levels of interluekin-2 in the corresponding cel groups were significantly lower than that in the untransfected cels (P 0.05). These findings indicate that CTLA4Ig-modified bone marrow stromal cels mediated by adenovirus can induce immune tolerance of T-lymphocyte from HLA haploidentical donors in vitro.
ABSTRACT
BACKGROUND:Imatinib resistance is a key issue in treatment of chronic myeloid leukemia. It is confirmed that the leukemia cels can obtain drug resistance phenotype mediated by adhesion to the bone marrow stromal cels (BMSCs). But, the role of BMSCs in imatinib resistance is unclear because chronic myeloid leukemia is deficient in adhesion function. OBJECTIVE: To in vitro simulate the bone marrow microenvironment of patients with chronic myeloid leukemia and to explore its influences on imatinib sensitivity as wel as possible mechanisms. METHODS:BMSCs isolated from patients with chronic myeloid leukemia but not undergoing treatment were co-cultured with K562 cels to construct the BMSCs-K562 cel co-culture model in chronic myeloid leukemia patients, then exposed to 0.2-3.2 μmol/L imatinib for 48 hours, and the proliferation inhibition rate of K562 cels was studied by MTT assay. The apoptosis rate of K562 cels and the expression of the CXCR4 in K562 cels exposed to 0.5 μmol/L imatinib for 72 hours were detected by flow cytometry. The K562 cels were exposed to 0.5 μmol/L imatinib for 4 hours and labeled by Calcein-AM fluorescent labeling sytem, and then, the adhesion rate of the K562 cels was calculated based on fluorescence intensity. RESULTS AND CONCLUSION: The suppressing effect of imatinib (0.2-3.2μmol/L) on the proliferation of K562 cels was weakened significantly by co-culture with the bone marrow stromal cels from patients with chronic myeloid leukemia. The apoptosis rate of K562 cels exposed to 0.5 μmol/L imatinib for 72 hours in co-culture group was significantly lower than that in the suspension culture group (P=0.020). The positive rates of CXCR4 in the co-culture group and suspension culture groups were both increased after exposure to 0.5 μmol/L imatinib for 72 hours (P=0.001). The adhesion rate of the K562 cels to the BMSCs was elevated from (32.18±6.17)% to (68.97±11.08)% when the K562 cels were exposed to 0.5 μmol/L imatinib for 4 hours, and the difference had statistical significance (P=0.022). These findings indicate that the co-culture with the BMSCs from patients with chronic myeloid leukemia can mediate K562 cels resistance to imatinib, which may be related to that the co-culture with BMSCs and exposure to imatinib can induce the K562 cels to express CXCR4. But the mechanism needs in-depth studies.
ABSTRACT
Objective To investigate influences of co-culture with the bone marrow stromal cells (BMSCs) on imatinib sensitivity,and the role of stromal cell-derived factor-1 (SDF-1)/chemokine receptor 4 (CXCR4) axis in imatinib resistance of K562 cells in the co-culture model.Methods The model was constructed by co-culturing K562 cells with BMSCs isolated and cultured from the patients with chronic myeloid leukemia.The apoptosis rate and the CXCR4 expressing rate of the K562 cells exposed to 0.5 μmol/L imatinib for 72 hours were detected by fluorescent-activated cell scanning (FACS) machine.The K562 cells were exposed to 0.5 μmol/L imatinib for 4 hours,and labelled by calckin-AM fluorescent labeling sytem.The adhesion rate of the K562 cells co-cultured with BMSCs for 24 hours was calculated with fluorescence intensity.The IC50 value of K562 cells exposed to imatinib was detected by methyl thiazolyl tetrazolium (MTT) assay while the SDF-1/CXCR4 axis was blocked by 10 μg/ml monoclonal antibody of CXCR4.Results The apoptosis rate of K562 cells exposed to 0.5 μmol/L imatinib for 72 hours in co-culture group and suspension culture group was (15.48 ±4.17) ％ and (32.01 ±6.83) ％,respectively.The apoptosis rates of K562 cells in the two groups were significantly different (t =5.587,P =0.001).For the co-culture group,the CXCR4 expressing rates of K562 cells unexposed and exposed to 0.5 μmol/L imatinib for 72 hours were (20.31 ± 3.76) ％ (suspension cultured:11.28％ ± 3.44％) and (53.64 ± 5.35) ％ (suspension cultured:25.34％ ± 3.21％),respectively.Those results showed that co-culture with BMSCs and exposure to imatinib induced the K562 cells to express CXCR4.The adhesion rates of the K562 cells to the BMSCs were elevated from (42.18 ± 6.17) ％ to (68.97 ± 11.08) ％ when the K562 cells were exposed to 0.5 μmol/L imatinib for 4 hours.The IC50 values of block group (the SDF-1/CXCR4 axis was blocked by 10 μg/ml monoclonal antibody of CXCR4) and unblock group were (0.68 ± 0.04) μmol/L and (1.27 ± 0.05) μmol/L,respectively.The IC50 values of two groups were significantly different(t =4.869,P =0.001).Conclusions The K562 cells co-cultured with the BMSCs from the patients with chronic myeloid leukemia can obtain resistance to imatinib,which was related with that co-culture with the BMSCs and exposure to imatinib can induce the K562 cells to express CXCR4.To a certain extent,the imatinib resistance mediated by co-culture with BMSCs can be reversed by monoclonal antibody of CXCR4.
ABSTRACT
Since 1988,our school has been enrolling in increasing numbers students in 7-year programs of clinical medicine and stomatology. During the period, we have accumulated some experience, which can be embodied in five major points. On this basis, we have, starting from this year, enrolled students working on doctoral degree in 8-year programs with candidates who have completed 4-year undergraduate programs of science and engineering, designed to further intensify reforms in medical education programs.