| Objective: To observe the clinical efficacy of Yiqiqingshihuayu therapy on pelvic inflammatory disease(pelvic,inflammatory,disease,PID)with repeated attacks(Qi deficiency,blood stasis and damp retention),and from the association between growth arrest specific protein6(growth,arrest,specific,gene,6,Gas6)and Toll like receptors 2(Toll-like,Receptor 2,TLR2)and Toll like receptor 4(Toll-like,receptor 4,TLR4)to investigate the mechanism of PID(repeated Qi and blood stasis and dampness syndrome)with Yiqiqingshihuayu therapy.Methods: 50 Patients with recurrent PID were selected as test group in the gynaecological clinic of the Chengdu University of Traditional Chinese Medicine between February 2016 and December 2016,25 healthy people were selected as control group,Patient in test group was treated for 3 months with the comprehensive method-yiqi qingshi huayu method(oral penyankangfu decoction,traditional chinese medicine enema,moxibustion(Zhongwan/Shenque/Guanyuan/Zusanli).Healthy people in control group were without any treatment.Recording the syndrome score of test group every two weeks and signs score every 4weeks.,the scores of TCM syndromes and clinical efficacy were observed before and after treatment for 12 weeks,the recurrence rate was evaluated after 12 weeks of withdrawal.The blood of test group were collected separately before and after the treatment,while the healthy group before the study.Making tests just as follows:Testing the rate of TLR2 and TLR4 expression in peripheral blood leukocytes by flow cytometry detection,detecting the expression level of TLR2 and TLR4 mRNA in peripheral blood mononuclear cells by real-time PCR and detecting the expression level of Gas6 by ELISA method.There had 15 cases of test group were picked up randomly for safety evaluation.Results:1.Clinical observation results:(1)Single symptom effect: the lower abdomen pain or bearing-down pain,lumbosacral pain,which could be worse during menstruation or being tired,much Leucorrhea,of which color could be white or yellow,getting tired easily,menostaxis or menorrhagia,dark menstrual blood or menstruation mixed with blood clot,chest stufly,anorexia,sticky and greasy in mouth,all of which were comparised as observing indexes before and after treatment,the difference hadstatistical significance(P<0.05).(2)Single sign effect: uterine is restricted to move and tenderness,left and right adnexa tenderness,left and right adnexa are thickening and enclosed mass,uterosacral ligament is thickening and tenderness,all of which were comparised as observing indexes before and after treatment,the difference had statistical significance(P<0.05).(3)Analgesic efficacy:After treatment,The cure rate of the experimental group was24%,Apparent efficiency 36%,Effective rate 34%,The total effective rate was 94%,The pelvic pain score of the test group was 4.80 + 1.40 before treatment,After treatment,it was 1.68 +1.10,It was decreased by 3.12 + 1.62 before treatment,The difference between the two groups was statistically significant(P<0.05).(4)Signs effect: After the treatment,the cure rate,the remarkable effect rate and effect rate were 2.00%,56.00%,38.00%,total effective rate were 96.00%.The signs score of the test group were 9.02±1.99 before the treatment,and after that were 3.60±1.83,falling 5.42±2.52 than before.the difference had statistical significance(P<0.05).(5)Curative effect of TCM syndrome: After the treatment,the cure rate,the remarkable effect rate and effect rate were 0.00%,74.00%,22.00%,total effective rate were 96.00%.The TCM syndrome score of the test group was 9.96±3.74 before the treatment,and after that was4.52±2.21,falling 5.44±1.53 than before,the difference had statistical significance(P<0.05).(6)Safety evaluation: There was no serious adverse events and three adverse events during the process of tests,of which 2 cases happened mild adverse reaction and there was no obvious abnormal for laboratory examination.(7)Recurrence rate: Evaluating the 30 patient's condition with PID recurrent after stoping the treatment for three months,2 cases were recurrent,28 cases weren't recurrent,the recurrence rate was 6.67%.2.Laboratory index evaluation:(1)Protein expression index: The rate expression of TLR2,TLR4 of two groups inmonocyte were59.80±21.39,32.17±21.16(testgroup)and45.99±18.91,23.70±16.76(control group),which were significantly higher than which in lymphocyte and granulocyte.And the level of test group were higher than that ofcontrol group,the difference of TLR2 had statistical significance(P<0.05),while TLR4 had no statisticant significance(P>0.05).Comparing the rate of TLR2?TLR4 expression before the test with after the test showed that the latter former was higher than the latter,the difference of TLR2 's expression in lymphocyte,monocyte and granulocyte had statistical significance(P<0.05),the difference of TLR4 's expression in monocyte had statistical significance(P<0.05),the difference of which in lymphocyte and granulocyte had no statistical significance(P>0.05).(2)Gene expression index: The comparison between two groups before observing showed that TLR2 ? TLR4 mRNA expression level of test group was higher than that of the control group,the difference between two groups had statistical significance(P<0.05).Comparing the expression of TLR2?TLR4 mRNA before the treatment with after the treatment showed that the expression of the former was lower than that of the latter,the difference had statistical significance(P<0.05).(3)Cytokines indicators: Comparing the two groups before observing discovered that,the expression level of peripheral blood Gas6 in test group were higher than which in control group,the difference had no statistical significance(P>0.05).Comparing the expression of it before the treatment with after the treatment showed that the expression of the former was higher than that of the latter,the difference had statistical significance(P<0.05).Correlation analysis: The expression of TLR2 protein and mRNA in the experimental group was positively correlated with the expression of TLR4 protein and mRNA,Correlation coefficients were equal to 0.348 and 0.622,The latter is more significant than the former;The expression of TLR2 protein and mRNA in the control group was positively correlated with the expression of TLR4 protein and mRNA,Correlation coefficients were equal to 0.483 and0.599,The latter is more significant than the former;There was a negative correlation between Gas6 and TLR4 protein,TLR2 and TLR4 mRNA expression in the experimental group,Correlation coefficients were equal to-0.175,-0.327 and-0.052,Among them,the correlation between Gas6 and TLR2 mRNA was significantly higher than the other two.The expression of Gas6 and TLR2,TLR2 and TLR4 mRNA were negatively correlated in the control group,Correlation coefficients were equal to-0.016,-0.091 and-0.19,The correlation was not significant.Conclusions:1.Yiqi Huayu Huayu comprehensive treatment of recurrent pelvic inflammatory disease in the improvement of pelvic pain symptoms,signs and symptoms of traditional Chinese medicine,the curative effect is accurate,after stopping the drug in March,the recurrence rate is low,no obvious adverse reactions.2.TLR2?TLR4 and Gas6 may participate in the process of recurrent PID.3.The therapeutic mechanism of yiqi qingshi huayu comprehensive method treat recurrent pelvic inflammatory disease may be related to reduce the expression of TLR2,TLR4 in lymphocyte,and Gas6 in peripheral blood.4..TLR2 and TLR4 may be positively related to recurrent immune responses in PID and play a coordinating role;Gas6 and TLR2 m RNA may be negatively related to recurrent immune responses in PID and play a reciprocal inhibitory role;Gas6 and TLR4,mRNA,TLR2,and TLR4 proteins may not be significantly related to the repeated immune response to PID. |
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