| ObjectiveTo evaluate the clinical efficacy of Replenishing qi,Expelling dampness and Dissipating blood stasis therapy of TCM in the treatment of recurrent pelvic inflammatory disease(repeated Qi and blood stasis and dampness syndrome),and to explore its regulation of Toll-like receptor 7(TLR7),Toll-like receptor 9(TLR9),myeloid differentiation primary response gene 88(MyD88)and interleukin-6(IL-6)expression related immune mechanisms.MethodsFrom August 2017 to May 2018,a total of 30 patients who met the RPID diagnosis were selected as the experimental group.All of them were from the gynecological outpatient clinic of the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine during this time period,and 10 healthy volunteers were selected as control group at the same period.The experimental group was given “Replenishing qi,Expelling dampness and Dissipating blood stasis therapy”,namely oral Chinese medicine(penyankangfu decoction),traditional chinese medicine enema(gynecological enema)and moxibustion treatment(Zhongwan,Shenque,Guanyuan and bilateral Zusanli),a total of 3 months of treatment.According to the follow-up status of the experimental group,the clinical syndrome performance and the pelvic signs were scored every 4 weeks.After the 12-week treatment period,the syndromes and signs were compared before and after treatment and assess clinical efficacy.The patients were followed up for 3 months after stopping the drug,and the recurrence of the experimental group was observed and evaluated.The healthy control group did not receive any treatment.The expedited experimental indicators were collected from the experimental group and the healthy control group at the time of enrollment and at the end of the treatment course.These include:(1)detection of TLR7 and TLR9 protein expression levels in peripheral blood lymphocytes and monocytes by flow cytometry;(2)Reverse Transcription Polymerase Chain Reaction(RT-PCR)detection of TLR7,TLR9 mRNA expression levels in peripheral blood mononuclear cells;(3)detection of serum MyD88 expression by enzyme-linked immunosorbent assay(ELISA);(4)detection of serum by chemiluminescence method IL-6 expression level.Twelve patients were randomly selected from the experimental group for safety evaluation.Results1.Clinical observations:1.1 Single symptom and physical efficacy: The single clinical symptoms of the experimental group were compared before and after treatment,including lower abdomen pain,lumbosacral pain,increased vaginal discharge,body fatigue,menstrual and menstrual changes,menstrual blood darkness,stagnation,greasy feelings,uterine activity,uterine tenderness,bilateral attachment thickening or tenderness,and thickening and tenderness of the palpebral ligament.The results showed that the differences were statistically significant(P<0.05).1.2 Analgesic efficacy: After treatment with this study protocol,the test group had a cure rate of 70%(recovery rate of 33.3%,significant efficiency of 36.7%),effective rate of 30%,and total effective rate of 100%.The pelvic pain scores before and after treatment in the experimental group were compared.The results showed that the pelvic pain score before treatment was 5.00±1.02,1.33±0.96 after treatment,which was 3.90±1.48 lower than that before treatment.The difference was statistically significant(P<0.01).1.3 Physical efficacy: After treatment with this study protocol,the test group had a cure rate of 70%(healing rate of 6.67%,significant efficiency of 63.33%),effective rate of 26.67%,and total effective rate of 96.67%.The total scores of the physical signs before and after treatment in the experimental group were compared.The results showed that the total score of the pre-treatment signs was 8.73±1.60,2.20±1.06 after treatment,and 6.76±1.84 before treatment.The difference was statistically significant(P<0.01).1.4 TCM syndrome efficacy: After treatment with this study protocol,the cure rate of the test group was 76.67%(recovery rate 0%,significant efficiency 76.67%),the effective rate was 20%,and the total effective rate was 96.67%.The total scores of syndrome efficacy before and after treatment in the experimental group were compared.The results showed that the total score of pre-treatment syndrome was 17.33±2.59,4.60±1.67 after treatment,which was 13.62±2.50 lower than that before treatment.The difference was statistically significant(P<0.01).1.5 Experimental group safety evaluation: A total of 1 study-related mild adverse reactions occurred during the study,and no serious adverse events occurred.No obvious abnormalities were observed in the laboratory tests.1.6 Recurrence rate: There was no recurrence of PID in the subjects during the treatment period.After the treatment,the patients were followed up to 12 weeks.The cumulative recurrence rate was 3.33%.2.Laboratory indicator test results:2.1 TLR7,TLR9 protein expression indicators: The positive expression rates of TLR7 and TLR9 in peripheral blood lymphocytes and monocytes of the experimental group and healthy control group were compared between the two groups.The results showed that the difference was statistically significant(P < 0.05).However,the expression rate of TLR9 in the experimental group increased,but there was no significant difference compared with the healthy control group(P>0.05).The positive expression rates of TLR7 and TLR9 in the experimental group were compared with those of the control group.The results showed that the expression rates of the two groups were lower than those before treatment,and the difference was statistically significant(P<0.05).2.2 TLR7,TLR9 mRNA gene expression indicators: The expression levels of TLR7 and TLR9 mRNA in peripheral blood mononuclear cells of the experimental group and healthy control group were compared between the two groups.The results showed that the difference was statistically significant(P<0.05).TLR9 mRNA was also highly expressed in the experimental group,but there was no significant difference compared with the healthy control group(P>0.05).The expression levels of TLR7 and TLR9 mRNA before and after treatment in the experimental group were compared by self-control.The results showed that the expression levels of the two groups were lower than those before treatment(P<0.05).2.3 Cytokine indicators: The expression levels of MyD88 and IL-6 in the peripheral blood of the experimental group and the healthy control group were compared between the two groups.The results showed that the experimental group was higher than the control group,the difference was statistically significant(P<0.05).The expression levels of MyD88 and IL-6 in peripheral blood of the experimental group before and after treatment were compared by self-control.The results showed that the expression levels of the two groups were lower than those before treatment(P<0.05).2.4 Correlation analysis: According to the test results,there was a positive correlation between TLR7 and TLR9 protein expression in peripheral blood mononuclear cells of the experimental group,the correlation coefficient was 0.878,and the correlation was statistically significant(P<0.01).Those two were also positively correlated with the expression of TLR7 mRNA,and the correlation coefficients were respectively = 0.547,0.457,and the correlation was statistically significant(P<0.05).The correlation between the former was significantly higher than that of the latter(P<0.01);the correlation between the other indicators was not statistically significant(P>0.05).Conclusion1.Replenishing qi,Expelling dampness and Dissipating blood stasis therapy of TCM is effective in improving the symptoms,signs and TCM syndromes of pelvic pain in patients with RPID(Qi deficiency,blood stasis and damp retention).After 3 months of withdrawal,the recurrence rate was low,and there was no obvious adverse reaction.2.The imbalance of immune homeostasis which was caused by TLR7,TLR9,MyD88,IL-6 may lead to RPID.3.The efficacy of Replenishing qi,Expelling dampness and Dissipating blood stasis therapy of TCM in the treatment of recurrent PID may be related to downregulation of TLR7,TLR9 expression,MyD88 and inflammatory factor IL-6 expression in peripheral blood mononuclear cells,inhibiting the release of downstream inflammatory factors and reducing Inflammatory response,improve immune function,and play a therapeutic role. |
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