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Objective:To investigate the effects of hyperthermia on the biological behavior of human laryngeal cancer Hep-2 cisplatin-resistant (Hep-2/CDDP) cell line and its possible mechanism.Methods:Hep-2/CDDP cell line was induced by high impact combined with increasing concentration method. Cell count method was used to detect the cell proliferation ability of Hep-2 parental cell group (Hep-2 cells without cisplatin-resistance and the cells were cultured with RPMI 1640 cultured medium without cisplatin), Hep-2/CDDP cell group and Hep-2/CDDP+cisplatin group (using RPMI 1640 cultured medium including 4 mg/L cisplatin). Hep-2/CDDP cell group and Hep-2 parental cell group were treated with cultured medium including 0, 0.004, 0.04, 0.4, 4, 40 mg/L cisplatin, respectively. The sensitivity of Hep-2/CDDP cells to cisplatin, vincristine and 5-fluorouracil was determined by using methyl thiazolyl tetrazolium (MTT) method. The half inhibitory concentration ( IC50) and resistance index (RI) were also calculated. Hep-2/CDDP cell group was divided into 4 subgroups: the cells in the control group were cultured for 24 h at 37 ℃; the cells in hyperthermia group were treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h; the cells in cisplatin group were cultured at 37 ℃ for 24 h in cultured medium containing 4 mg/L cisplatin. The cells in hyperthermia combined with cisplatin group were cultured in cultured medium containing 4 mg/L cisplatin, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. The effects of hyperthermia combined with cisplatin on the proliferation and early apoptosis of Hep-2/CDDP cells were detected by using MTT and flow cytometry. The interaction of hyperthermia combined with cisplatin on the proliferation and early apoptosis of HEP-2/CDDP cells was observed by using factorial analysis. Western blotting was used to detect the effect of hyperthermia combined with cisplatin on the expressions of wild-type p53 and PI3K in Hep-2/CDDP cells. Hep-2/CDDP cells were divided into 4 groups: the control group (Hep-2/CDDP cells were cultured for 24 h at 37 ℃); chemotherapy group was treated with 12 mg/L vincristine or 9 mg/L 5-fluorouracil; in the hyperthermia group, Hep-2/CDDP cells were treated at 43℃ for 2 h and then re-cultured at 37 ℃ for 22 h; in hyperthermia combined with chemotherapy group, the cells were cultured in a medium containing 12 mg/L vincristine or 9 mg/L 5-fluorouracil, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. MTT method was used to detect the effect of hyperthermia combined with vincristine and 5-fluorouracil on the proliferation of Hep-2/CDDP cells. Results:Hep-2/CDDP cell line was successfully established. There were no significant differences in the number of cells in Hep-2/CDDP cell group, Hep-2 parental cell line group and Hep-2/CDDP + cisplatin cell group at different time points (all P > 0.05), and the doubling time was 43.8, 40.6 and 43.5 h, respectively. The IC50 of Hep-2 parental cell line group and Hep-2/CDDP cell group to cisplatin was 4.771 mg/L and 42.749 mg/L, respectively, and the RI was 8.960. Hyperthermia combined with cisplatin could inhibit the proliferation of Hep-2/CDDP cells ( F = 327.91, P < 0.05) and promote the early apoptosis of Hep-2/CDDP cells ( F = 724.63, P < 0.05). Factorial analysis showed that hyperthermia combined with cisplatin had an interaction effect on the proliferation and early apoptosis of Hep-2/CDDP cells ( F = 185.68, 472.51, all P < 0.05). Western blotting showed that the relative expression levels of wild-type p53 protein and PI3K protein in the control group, hyperthermia group, cisplatin group and hyperthermia combined with cisplatin group were significantly different ( F = 547.76, 404.44, all P < 0.01). Hyperthermia combined with vincristine or 5-fluorouracil could inhibit the proliferation of Hep-2/CDDP cells ( F = 33.06, 34.61, all P < 0.05). Factorial analysis showed that hyperthermia combined with vincristine and 5-fluorouracil had no interaction effect on the proliferation of Hep-2/CDDP cells ( F = 0.64,0.60, all P > 0.05). Conclusions:Hyperthermia may reverse the resistance of Hep-2/CDDP cell line to cisplatin by upregulating wild-type p53 expression and inhibiting the PI3K pathway. Hep-2/CDDP cell line has cross-resistance to vincristine and 5-fluorouracil. Hyperthermia can increase the sensitivity of Hep-2/CDDP cell line to vincristine and 5-fluorouracil.
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According to the difference of menstrual quantity, color, quality and the symptoms, we can distinguish two types of menstrual disorders: hemorrhagic and impotency disorders. In this paper, we summarized the clinical progress of Traditional Chinese Medicine in treating menstrual disorders from the following areas: the flexible use of Chinese patent medicine, treatment based on syndrome differentiation and syndrome, the recognize of the artificial cycle in modern medicine, and non-drug therapy.
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Objective:To investigate the change of apoptosis-associated genes in human lung adenocarcinoma cell lines A549/DDP cells,which were induced by the recombinant human interleukin-24(rhIL-24) combined with Cisplatin (DDP). Methods: Six genes expression level by GeXP genetic analysis system at the same time,after rhIL-24,DDP and rhIL-24+DDP were used to intervene in A549/DDP cells. Results:rhIL-24 could induce Bax gene,Caspase3 gene and Rb gene transcription up regulation;Bcl-2 gene and survivin gene transcription down regulation. But no regular change in the genes expression level of P53. Bax,Survivin and Rb were more obviously changed after rhIL-24 combined with DDP. Conclusion: RhIL-24 can induce apoptosis of A549/DDP cells through upregulated the genes expression level of Bax,Caspase3,Rb and down regulated of Bcl-2,Survivin.
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Objective:Adjuvant arthritis(AA)rats interfered with recombinant hIL-10,methotrexate(MTX)separately,we detected the changes of T cell subsets in rat′s blood and IL-17 A in rat′s serum.Methods:AA rats interfered with recombinant hIL-10, MTX and IL-10 plus MTX.The changes of the CD4+and CD8+T cell subsets were detected by flow cytometry;the Levels of IL-17A in serum of rats were measured by ELISA method.Results:Compare to normal control group ,the CD4+T cell total quantity and CD4+/CD8+T cell Proportion was significantly decreased , there were significant differences between the treatment group and the untreated group(P<0.05 ), the IL-17A of treatment group was significantly decreased , there was a significant differences ( P<0.05 ).The combination group had significant difference compared with IL-10 group.Conclusion:Recombinant hIL-10 can down-regulate the blood of AA rats in the number of CD4 +T cells and CD4+/CD8+ratio,reduce serum levels of IL-17A.The combination group compared with the IL-10 group effect on serum IL-17A more significantly,the results displayed recombinant hIL-10 plus MTX in the treatment of rheumatoid arthritis can better play the role.
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Objective:To elucidate the characterization of CD8+T cell in H1N1 influenza vaccine for children.Methods:PBMCs were isolated from 31 children aged from 3 to 6 years old who had accepted H 1N1 influenza vaccine during December 2009 to January 2010.The lymphocytes were joined with the H 1N1 influenza vaccine as experimental group and cultured .The experiment set without vaccine group as control group .At last we detected the surface molecules by FCM .The CCK-8 assay was added to detecting cellular proliferation and cellular proliferation index were detected by CCK-8.Results: CD8+T cells of PBMC in the two groups were 13.41%and 9.41%,P>0.05.CD8+CD45RAA+naive T cells in the two groups were up to more than 80%,P>0.05.The proportion of CD8+CD45ROA+memory T cells in two groups were up to 17%-19%,P>0.05.Two subsets of CD8+CD45ROA+memory T cells :CCR7+and CD62L+single positive memory T cell subsets in the experimental group were significantly lower than that of the control group,P<0.05.The CCK-8 assay was added to detect cellular proliferation .Only 51.16% of which cellular proliferation index was greater than 0.8,with none was greater than 1 in this study.Conclusion:This study showed that the CD4+T cells were low-level,naive T cells (CD8+CD45RAA+)were higher,with antigen stimulation and response.H1N1 vaccination specific memory T cells were few in number , specific memory T cell subsets were diversity , control memory cells were the main phenotypic characteristics .Cellular proliferation index showed that the proliferation of specific CD 8+T cells vaccine was poor .
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OBJECTIVE: To discuss the distribution laws of traditional Chinese medicine (TCM) syndrome factor and their combination in coronary heart disease (CHD), and to study the correlation between the TCM syndrome factor combination and cardiac function as well as blood-lipid. METHODS: The parameters of the cardiac function of 300 patients with a final diagnosis of CHD by coronary angiography were measured by echocardiography, and the levels of blood lipids in the CHD patients were detected. An analysis of the correlation was done between the TCM syndrome factor combination and cardiac function as well as blood-lipid in CHD. RESULTS: The TCM syndrome factor combinations of CHD were blood stasis due to qi deficiency, qi and yin deficiency, intermingled phlegm and blood stasis, and yang deficiency and blood stasis. The ejection fraction of CHD patients with yang deficiency and blood stasis was markedly decreased. The levels of triglyceride and low-density lipoprotein cholesterol in CHD patients with intermingled phlegm and blood stasis were markedly increased, and the level of triglyceride in CHD patients with qi and yin deficiency was markedly increased too. CONCLUSION: The treatment of CHD should aim directly at the symptoms and causes. It is also proved that some compound traditional Chinese herbal medicines for supplementing qi and activating blood circulation, nourishing yin and resolving phlegm, and activating yang should be used in treatment of CHD. In cases of CHD with low cardiac function, particular emphasis should be laid on activating yang and blood circulation, while in cases of CHD with blood-lipid disturbance, particular emphasis should be laid on resolving phlegm and activating blood circulation, replenishing qi and nourishing yin.
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Syndrome factors and their combinations based on syndromes are the important links of construction of syndrome differentiation system,in which the theory of prescriptions corresponding to syndromes plays a key role.The basic requirements of good clinical eff icacy are'syndrome factors'seen as the basic connotation of'syndromes'in prescriptions corresponding to syndromes,keeping the compatibility of effect classical prescriptions,emphasizing further detailed formula indication and dynamic evolution,and increasing the conformity of prescription and syndrome.These are also the basis of construction of syndrome differentiation system.Method of evidence-based medicine should be used for reference in clinical research design,and reverse thinking of conjecturing syndrome based on prescription be used for guidance in the constructing process.
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Objective To explore the correlation between syndrome elements of traditional Chinese medicine (TCM) and quality of life (QOL) of the patients with angina pectoris.Methods With the clinical epidemiology method, 233 patients with angina pectoris confirmed by coronary angiography were collected, and their QOL were assessed by the Seattle Angina Questionnaire (SAQ). The correlation of the scores of five different dimensions, including somatic movement limitation, angina stability and attacks, treatment satisfaction, and disease recognition, and the total scores of SAQ and the TCM syndrome elements was analyzed by Spearman rank correlation analysis.Results The six factors, namely, qi stagnation, blood stasis, phlegm turbidity, stagnated heat, yang and yin deficiency, had the correlation with one or more of the dimensions and total score of QOL, in which the phlegm turbidity had a negative impact (P