| Objective1To observe the clinical efficacy of Chinese herbal compound of Cangbaishiduqing (CBSDQ) in treating Mycoplasma cervicitis (the Syndrome of qi deficiency caused by Dampness Dampness toxin).2To observe the effect of Cangbaishiduqing intervening the establishment of ureaplasma urealyticum model in BALB/c mice.3To probe the possible immunological regulation mechanism of Cangbaishiduqing intervening ureaplasma urealyticum model in BALB/c mice from IL-2level in serum, IL-12level and LTC4level of vaginal mucosa tissue, the expression of C3b receptors on vaginal mucosa tissue.MethodThe research contains two parts:I The clinical studyBy the study method of randomized double-blind controlled.Processed the Cangbaishiduqing granules and Relinqing (RLQ) granules into the same outer package and the harmonized compiled blind. Selected patients of mycoplasma cervicitis were randomly dispensing granules by visiting sequence. During the treatment, the patients were taking granules one bag, twice a day in the course of two weeks. Recorded the symptom score before treatment and after two weeks of medication. Detected vaginal pll, count while blood cell (WBC) of cervical secretions, and cultivated mycoplasma before treatment, after two weeks of medication, drug withdrawal one week, three weeks. Determined the efficacyultimately. In the end of study, the blind was exposed.2The experimental studySeventy. BALB/c mice were randomly divided into groups A1-1, A1-2, B1, A2-1A2-2, B2and C, ten mice per group.2.1Experiment I2.1.1After7days, low dose of Cangbaishiduqing intervening in Group A1-1, high dose intervening in group Al-2, no intervention in group B1, three groups were established ureaplasma urealyticum model. At the end of the experiment, collecting vaginal secretions of each mice were cultivated UU, to observe the effect of Cangbaishiduqing intervention in Ureaplasma urealyticum colonization by calculated the positive rate of UU.2.1.2Probed the possible immunological regulation mechanism of Cangbaishiduqing intervening the establishment of ureaplasma urealyticum model in BALB/c mice.①By ELISA, three groups were detected blood Interleukin-2(IL-2) in the end of the experiment.②By ELISA, three groups were detected the content of interleukin-12(IL-12) and leukotriene C4(LTC4) in vaginal mucosa tissue.③By immunohistochemistry, C3b receptors on vaginal mucosa tissue were dyed. Chose meaningful images, each case measured5horizons, and calculated the average optical density (MOD) of the positive reaction by application of computer image processing system by computer.2.2Experiment II2.2.1First, Group A2-1, Group A2-2, Group B2were established ureaplasma urealyticum model in four weeks. Secondly, low dose of-Cangbaishiduqing was intervening in Group A2-1, high dose intervening in group A2-2, no intervention in group B2for7days. At the end of the experiment, collecting vaginal secretions of each mice were cultivated UU, to observe the effect of Cangbaishiduqing intervention in Ureaplasma urealyticum colonization by calculated the positive rate of UU.2.2.2Probed the possible immunological regulation mechanism of Cangbaishiduqing treating the establishment of ureaplasma urealyticum model in BALB/c mice①By ELISA, three groups were detected blood IL-2in the end of the experiment.②By ELISA, three groups were detected the content of IL-12and LTC4in vaginal mucosa tissue.③By immunohistochemistry, C3b receptors on vaginal mucosa tissue were dyed. Chose meaningful images, each case measured5horizons, and calculated MOD of the positive reaction by application of computer image processing system by computer.2.3Group C was the blank group. Group C did not enter the experiment Ⅰ or Ⅱ. At the end of experiment Ⅰ and Ⅱ, collecting vaginal secretions of each mice in group C were cultivated UU with other groups at the same time. Each mouse in group C was detected the same indicators.Results1The clinical study1.166patients were selected. In the33cases of CBSDQ group,14cases were cured,6cases shew effects,5cases had maken progress,8cases shew no effect, the marked recovery rate was60.6%; in the33cases of RLQ group,8cases were cured,4cases shew effects,6cases had maken progress,15cases shew no effect, the marked recovery rate was36.4%. The difference was significant between these two groups (P<0.05). The marked recovery rate of CBSDQ group in the treatment of mycoplasma cervicitis was higher than the rate of RLQ group. The efficacy of CBSDQ was better than the efficacy of RLQ in treatment of mycoplasma cervicitis (the Syndrome of qi deficiency caused by Dampness Dampness toxin).1.2The vaginal pH value went down to normal gradually in CBSDQ group. The vaginal pH value went down but not to normal in RLQ group. The difference was prominent after and before treated (P<0.05). The vaginal PH value in CBSDQ group decreased more significantly than in RLQ group.1.3The WBC counts of cervical secretions went down in the two groups. The difference in the WBC counts was prominent after and before treated (P<0.01). The WBC counts in CBSDQ group decreased more significantly (P<0.01) than in RLQ group. 1.4In improving the main symptoms, the difference was prominent (P<0.05) after and before treated in two groups. The syndrome score in CBSDQ group decreased more significantly than in RLQ group.2The experimental study2.1The pathogenicity of ureaplasma urealyticum serotype4In experiment II,29BALB/c mice were successfully established ureaplasma urealyticum model in four weeks. The vaginal secretions of each mouse were cultivated UU all positive (100%). Ureaplasma urealyticum serotype4could cause vaginal infections in BALB/c mice。2.2Successfully established the ureaplasma urealyticum model of vaginal infections in BALB/c miceEstrogen-treated female BALB/c mice were inoculated intravaginally by UU serotype4. UU serotype4was successfully inoculated. This method was successfully established the ureaplasma urealyticum model of vaginal infections in BALB/c mice.2.3Excellent experimental QC conditions.The positive rate of UU in group C was0%before and after experiment. The experimental QC conditions were excellent.2.4The success rate of UU colonizationAt the end of experiment I and II, the positive rate of UU of vaginal secretions respectively in group A1-1, group A1-2, group B1, group A2-1, group A2-2, group B2was70.00%(7/10),66.67%(6/9),90.00%(9/10),66.67%(6/9),70.00%(7/10),100.00%(10/10)In experiment I, some mice in group A1-1and group A1-2intervened by Cangbaishiduqing were not successfully established the ureaplasma urealyticum model.In experiment II, all the mice of group A2-1and group A2-2were successfully established the ureaplasma urealyticum model. After treaded by Cangbaishiduqing, UU was negative in some mice. But the difference of the positive rate of UU was inapparent between groups (P>0.05).2.5IL-2level in serumAt the end of experiment I and II, the values of IL-2in seven groupse were in descending order:group A1-1(54.16±1.52), group A1-2(51.86±1.51), group A2-1(45.33±1.82), group A2-2(44.45±2.75), group C(44.02±2.14), group B1(41.29±1.35), group B2(35.40±1.78).The values of IL-2in group B1and group B2were lower than the values in group C. The difference was prominent (P<0.05). The blood IL-2values were in. low level in UU infection of BALB/c mice.In experiment I, the values of IL-2were raised in group A1-1and group Al-2intervened by Cangbaishiduqing. The values of IL-2in group A1-1and group A1-2had prominent difference with the values in group B1and group C (P<0.05). Cangbaishiduqing played an important role in the prevention of UU infection by raising the serum IL-2levels.In experiment II, all the mice of group A2-1and group A2-2were successfully established the ureaplasma urcalyticum model. After treaded by Cangbaishiduqing, the values of IL-2were raised in both group A2-1and group A2-2. The values of IL-2in group A2-1and group A2-2had prominent difference with the values in group B2(P<0.05),which had inapparent difference with the values in group C(P>0.05). Cangbaishiduqing played an important role in the anti-UU infection by raising the serum IL-2levels.Cangbaishiduqing raised the blood IL-2levels in mice, increased TH1cells and activated or mobilized humoral immune function. Cangbaishiduqing played an important role in the prevention of UU infection and the anti-UU infection by raising the serum IL-2levels.2.61L-12in vaginal mucosa tissueAt the end of experiment I and II, the values of IL-12in seven groups were in descending order:group B2(19.37±14.22), group B1(18.41±13.23), group A2-2(12.52±3.96), group A2-1(12.06±4.92), group A1-1(9.32±2.88), group C (9.21±5.92), group A1-2(7.52±3.25).The values of IL-12in group B1and group B2were higher than the values in group C. The difference was prominent (P<0.05). The values of IL-12in vaginal mucosa tissue were in high level in UU infection of BALB/c mice.In experiment I, the values of IL-12were decreased in group A1-1and group A1-2intervened by Cangbaishiduqing. The values of IL-12in group A1-1and group A1-2had prominent difference with the values in group B1(P<0.05), which had inapparent difference with the values in group C(P>0.05). Cangbaishiduqing laycd an important role in the prevention of UU infection by decreasing the IL-12level of vaginal mucosa tissue.In experiment Ⅱ, all the mice of group A2-1and group A2－2were successfully established the ureaplasma urealyticum model. After treaded by Cangbaishiduqing, the values of IL-12were decreaseed in both group A2-1and group A2-2. The values of IL-12in group A2-1and group A2-2had inapparent difference with the values in group B2and group C (P>0.05). Cangbaishiduqing decreasing the IL-12high level was not significant.Cangbaishiduqing decreased the IL-12level of vaginal mucosa tissue in mice. Cangbaishiduqing played an important role in the prevention of UU infection by decreasing the serum IL－2. Cangbaishiduqing played an unimportant role in the anti-UU infection.2.7LTC4in vaginal mucosa tissueAt the end of experiment I and Ⅱ, the values of LTC4in seven groups were in descending order:group B2(485.61±387.36), group B1(399.88±297.61), group A2-1(288.54±113.48), group A2-2(244.91±52.65), group Al-1(198.90±54.36), group C (172.71±75.44), group A1-2(162.77±66.84).The values of LTC4in group B1and group B2were higher than the values in group C. The difference was prominent(P<0.05). The values of LTC4in vaginal mucosa tissue were in high level in UU infection of BALB/c mice.In experiment I, the values of LTC4were decreased in group A1-1and group A1-2intervened by Cangbaishiduqing. The values of LTC4in group A1-1and group A1-2had prominent difference with the values in group B1(P<0.05), which had inapparent difference with the values in group C(P>0.05). Cangbaishiduqing played an important role in the prevention of UU infection by decreasing the LTC4level of vaginal mucosa tissue.In experiment Ⅱ, all the mice of group A2-1and group A2-2were successfully established the ureaplasma urealyticum model. After treaded by Cangba i sh i duq i ng, the values of LTC4decreaseed in both group A2-1and group A2-2. The values of LTC4in group A2-1and group A2－2had prominent difference with the values in group B1(P<0.05), which had inapparent difference with the values in group C (P>0.05). Cangbaishiduqing played an important role in the anti-UU infection by decreasing the LTC4level of of vaginal mucosa tissue.’Cangbaishiduqing decreased the’LTC4level of vaginal mucosa’tissu’e in mice, controled vascular permeability, reduced inflammatory exudate, kept the integrity of vaginal mucous membrane barrier in order to greatly reduce the UU invasion.Cangbaishiduqing played an important role in the prevention of UU infection and the anti-UU infection by decreasing the LTC4level.2.8The expression of C3b receptors on vaginal mucosa tissueAt the end of ex·periment I and Ⅱ, the MOD of C3b receptors in seven groups were espectively:group A1-1(0.038±0.013), group Al－2(0.045±0.087), group A2-1(0.039±0.011), group A2-2(0.042±0.009), group B1(0.041±0.009), group B2(0.042±0.012), group C (0.042±0.010). The difference in seven groups was inapparent(P>0.05). Cangbaishiduqing might not play an immunomodulatory effects by regulating C3b receptors.ConclusionsThe efficacy of Cangbaishiduqing was better than the efficacy of Relinqing in treatment of mycoplasma cervicitis(the syndrome of qi deficiency caused by dampness toxin). Cangbaishiduqing could effectively decrease the vaginal pH value to normal levels and could decrease the WBC counts in order to keep the integrity of vaginal mucous membrane barrier. Cangbaishiduqing could significantly improve the systemic and local symptoms. Cangbaishiduqing played a strong role of anti-mycoplasma.By Cangbaishiduqing intervening, some mice were not successfully established the ureaplasma urealyticum model. Successfully established the ureaplasma urealyticum model after treaded by Cangbaishiduqing, UU was negative in some mice. Cangbaishiduqing could raise the blood I L-2level, decrease the IL-12level and the LTC4level of vaginal mucosa tissue, and might not play an immunomodulatory effects by regulating C3b receptors. Cangbaishiduqing played an important role in the prevention of UU infection and in the anti-UU infection. The innovation of this project, containing firstly the blood IL-2level, the IL-12level and the LTC4level of vaginal mucosa tissue, regulating C3b receptor,was to explore the mechanism of herbal compound from the prevention of UU infection and the anti-UU infection. |
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