| Part one:Experimental Study of GanDuQing Granules on Immune System in Cyclophosphamide-induced immunesuppressive miceObjective: To investigate the immune regulation effects of GanDuQingGranules in immunosuppressive mice, the immunosuppressive mice model was established to observe the impact on Immune System of GanDuQing Granules in Immune Organ Index, immune cell function, immune molecules content of immunosuppressive mice, and to explore its mechanisms.This study provide a theoretical basis for its clinical application.Methods:1. SPF grade Kunming mouse60, weight18-22g, were randomly divided into six groups, as follows:Normal control group(lavaged Physiological saline,7consecutive days); Cyclophosphamide model group (Lavaged saline0.2mL/10g, seven days, at the same time abdominal subcutaneous injected cyclophosphamide50mg/kg for3days); Positive control group (intraperitoneal subcutaneous injected cyclophosphamide50mg/kg modeling3consecutive days, then lavaged YuPingFeng Granules suspension2.25g/kg7consecutive days);GanDuQing Granules high group、middle grope、low dose group (intraperitoneal injected cyclophosphamide50mg/kg modeling daily3consecutive days, administered GanDuQing Granules8.1g/kg,4.05g/kg,2.025g/kg,7consecutive days). After the last administration, mouse were fasted12h, the eye was removed to collect blood, using flow cytometry to detect the proportion of NKT cells in the blood.2. To make model was the same as the first one. After blood collection from eyes, all mouse were killed to take thymus and spleen, which weighed to calculate the thymus and spleen index. Thymus was made to thymic tissue homogenate after centrifugation, and to detect the supernatant of IFN-y、IL-2、IL-4.3. To make model was the same as the first one. The mouse was killed to take its spleen, and the spleen was set into liquid nitrogen immediately, and subsequently transfered to-80℃cryogenic refrigerator, using immunohistochemical methods to detect the protein content of T-Bet and GATA-3in the spleen.Results:1.Compared with the normal control group, the thymus index and spleen index of cyclophosphamide model group was decreased significant(P<0.05), after the administration,the comparison between GanDuQing Granules dose group, YuPingFeng Granules control group, model group has no significant difference (P>0.05).2.Compared with normal control group, the content of NKT in the cyclophosphamide model group was obviously higher than normal control group, which has significant difference (P<0.05); after administration, compared with model group, NKT cells of Peripheral blood in GanDuQing high dose group was obviously higher than cyclophosphamide model group, which has significant difference(P<0.05); Compared with cyclophosphamide model group, the middle dose group of GanDuQing and YuPingFeng Positive control group was also increased, but the difference has no statistical significance(P>0.05).3.Compared with the normal control group, the content of IFN-y, IL-2in the cyclophosphamide model group was significantly lower than the control group, which has significant difference (P<0.05); after administration,the content of BFN-y in every dose group thymus tissue homogenate was significantly higher than the cyclophosphamide model group, the difference has statistical significance (P<0.05).The content of IL-2in the high dose group thymus tissue homogenate was significantly higher than the cyclophosphamide model group, the difference has statistical significance(P<0.05).Compared with the normal control group, the content of IL-4in the cyclophosphamide model group was significantly higher than the normal control group, the difference has statistical significance (P<0.05); after administration, the content of IL-4in the high and middle dose group thymus tissue homogenate was significantly lower than the cyclophospamide model group, the difference has statistical significance (P<0.05).4. Compared with the normal control group, the content of T-bet in the spleen of the cyclophosphamide model group was significantly lower than the control group, the difference has statistical significance (P<0.05); after administration, the content of T-bet in the high and middle dose group of GanDuQing Granules, YuPingFeng Positive control group was ignificantly higher than the cyclophosphamide model group, the difference has statistical significance(P<0.05). Compared with YuPingFeng Positive control group, the difference of the content of T-bet in the high dose group of GanDuQing has no statistical significance (P>0.05). It proved that GanDuQing Granules can improve the content of T-bet in the spleen tissue in immunosuppressed mice, and the effect was the same as YuPingFeng Granules.Compared with normal control group, the content of GATA-3in the spleen of the cyclophosphamide model groups was significantly higher than the normal control group, the difference has statistical significance (P<0.05); after administration, the content of GATA-3in the spleen of all GanDuQing dose group and YuPingFeng Positive control group was significantly lower than cyclophosphamide model group, and the difference was significant (P<0.05). Compared with YuPingFeng Positive control group, the difference of the content of GATA-3in the high dose group of GanDuQing has no statistical significance (P>0.05). It indicted that GanDuQing Granules can reduce the content of GATA-3in the spleen tissue of immunosuppressed mice, and the effect was the same as YuPingFeng Granules.5. Effects of GanDuQing Granules on Immune System were relative with dose, and the high and middle dose group was better than the low dose group.Conclusion:1.Using cyclophosphamide to make immunosuppressed mice model, which leaded to the thymus index, spleen index decreased, indicate that using cyclophosphamide to make immunosuppressive model was successful.2. GanDuQing Granules can improve the content of NKT cells in the peripheral blood of immunosuppressed mice, so that the body’s inherent immune can recover to the normal standard.3. GanDuQing Granules can improve the content of IFN-γand IL-2, decrease the content of IL-4in immunosuppressed mice; and balance cytokines of TH1/TH2subsets, regulate the content of immune cells; its effect was the same as YuPinFeng Granules. 4. GanDuQing Granules can regulate proportion of Thl and Th2, which may be partly due to the effect of transcription factor T-bet and GATA-3.Part two: Clinical observation of GanDuQing Granules to cure acute upper respiratory tract infection (Pattern of Qi Deficiency and Evil Invaded) and Clinical research on immune function in patientsObjective: Through clinical observation, to explore the clinical effect of GanDuQing Granules on treating acute upper respiratory tract infection and its impact on the content of T cells, B cells, NK cells from peripheral blood in patients. The study would demonstrate effectiveness and feasibility of GanDuQing Granules on treating acute upper respiratory tract infection (Pattern of Qi Deficiency and Evil Invaded), and provide a theoretical basic for clinical medicine, develop a new treatment of acute upper respiratory tract infection.Methods:In December2012to March2013, to collect39clinical patients with acute upper respiratory tract infection, which see a doctor in attachment hospital of ChengDu University of TCM, ChengDu Xi’an Road QingYang Community health service station. All the patients were divided into the treatment group (26patients) and the control group (13patients).The treatment group had been treated with GanDuQing Granules, the control group had been treated with anti-viral Granules, then made clinical observation of treatment for three days. To select11cases from the treatment group randomly and11cases from the control group randomly, each of them adopted flow cytometry to detect the percentage of T cells, B cells, NK cells in the peripheral blood of patients at the same day and3day latter, then compared two group.Results:1. Compared with control group, the cure and markedly cure rate of GanDuQing treatment group have significant difference(p<0.05).The cure rate of GanDuQing group was87.5%, the total effective rate was100%; the cure rate of the anti-viral control group was27.3%, the total effective rate was100%. The cure rate of GanDuQing Granules treatment group was significantly higher than the anti-viral control group, the difference was statistically significant (p<0.05).2. After treatment with GanDuQing Granules and anti-viral Granules, the main symptoms of two groups such as fever, sore throat, malaise and fatigue, shortness of breath were all significantly alleviated. Compared with3days ago, the curative effect of GanDuQing treatmen group have significant difference (P<0.05). Compared with3days ago, the curative effect of Anti-viral treatment group, such as the fever, sore throat have significant difference (P<0.05), but malaise, fatigue, shortness of breath was no significant difference (P>0.05). It proved that GanDuQing Granules can alleviate the main symptoms of cold such as fever, sore throat, fatigue, shortness of breath, and GanDuQing Granules were more effective than anti-viral Granules.3. After3days treatment, the secondary symptoms of cold of two groups such as fear of cold, poor sweating, headache, nasal congestion, runny nose, cough were also alleviate. Compare with treatment ago, the secondary symptoms of cold were significant difference after GanDuQing treatment (P<0.05). And Compared with treatment ago, the secondary symptoms of cold such as poor sweating, headache, nasal congestion, cough were more significant difference (P<0.05) after Anti-viral drugs treatment, but the symptoms of bad cold, runny nose were no significant difference (P>0.05). The effective effect of GanDuQing Granules to alleviate the secondary symptoms of a bad cold such as poor sweating, headache, nasal congestion, runny nose, cough were more effective than anti-viral Granules.4. Compared with GanDuQing treatment ago, the percentages of T cells, B cells, NK cells from patient blood had a clear upward trend after treatment, and the ratio of CD4/CD8had a more significant difference(P<0.05). Compare with anti-viral treatment ago, the content of T cells did not change significantly, and there was an upward trend in the percentage of B cells, NK cells after anti-viral treatment, but there was no significant difference (P>0.05). The percentage of T cell subsets, B cells, NK cells between two groups was not significant difference (P>0.05).Conclusion:GanDuQing Granules had a significant effect on curing acute upper respiratory tract infection symptoms such as fever, sore throat, fatigue, shortness of breath, and relieve secondary symptoms such as fear of cold, poor sweating, headache, nasal congestion, runny nose, cough, etc. The therapeutic effect of GanDuQing Granules was better than anti-virus Granules. GanDuQing Granules can increase the content of immune cells such as T cell subsets, B cells, NK cells and other cells in upper respiratory tract infection patient’s blood, enhance their immunity. GanDuQing Granules had a better therapeutic effect on curing acute upper respiratory tract infection (Pattern of Qi Deficiency and Evil Invaded), which can relieve a variety of cold symptoms significantly. At the same time, GanDuQing Granules can improve the immune function of patient with the upper respiratory tract infection. |
PDF Full Text Request