Clinical study of Cangbai Shiduqing Granul in reversing drug resistance of ureaplasma urealyticum

Zhng Yingchun(张颖纯), Li Yunwen(李元文), Li Wei(李纬), Zhou Zhiqing(周志强),Zho Yjing(赵雅静), Wu Meicho(吴美超), Shi Jingwen(石静纹),nd Ci Lingling(蔡玲玲)＊

a: Skin disease Hospital of Dalian, Liaoning Province, Dalian 110060, China

b: Dongfang Hospital, the Second Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100078, China

Clinical study of Cangbai Shiduqing Granul in reversing drug resistance of ureaplasma urealyticum

Zhang Yingchun(张颖纯)a, Li Yuanwen(李元文)b, Li Wei(李纬)b, Zhou Zhiqiang(周志强)b,Zhao Yajing(赵雅静)b, Wu Meichao(吴美超)b, Shi Jingwen(石静纹)b,and Cai Lingling(蔡玲玲)b＊

a: Skin disease Hospital of Dalian, Liaoning Province, Dalian 110060, China

b: Dongfang Hospital, the Second Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100078, China

UreaplasmaUrealyticum (UU) can not only cause a wide range of nongonococcalinfectionsto urogenital tract, but also lead to infertility, sterility, intrauterinefetal infection, abortion or other bad consequences. For males, UU may cause prostatitis,epididymitis and other diseases. With great harm to the human health, UU infection has been taken as one of the most important STDs in China for prevention and cure. However, the drug resistance of UU is increasingly stronger with the widespread use of antibiotics. Many nongonococcal patients with mycoplasma infection often fail to be cured after a long time of treatment,and drug sensitive tests find that, multidrug resistance brings physical and mental pains to patients all the time. Luckily,integrated Chinese traditional and western medicine greatly improves the cure rate, though the mechanism is not clear. There have been many studies on the reversion action and mechanism of TCM on the antibiotic resistance of bacteria, but studies on the reversion to UU drug resistance are deficient. Here we chose some voluntary patients with multidrug resistance as the research objects, who received treatment in my hospital in recent 3 years.They were given the oral administration of self-prescribed Cangbai Shiduqing Granule. Then, Cangbai Shiduqing Granule’s reversion effect on UU drug resistance, namely, the sensitivity transformation situation of drug-resistant antibiotics, was observed. And the observation results are presented in the paper.

Ureaplasmaurealyticum (uu); Drug resistance;Reversion; Antibiotics

MATERIAL AND METHOD

General data

Patients with nongonoccus vaginitis (cervicitis)and urethritis in the Dermatology and Venereology Department and the Gynecology Clinic of our hospital were chosen as research subjects. 51 voluntary patients,20 females and 31males, were screened out. They had antibiotic resistance, with cultured mycoplasma shown positive in the urethral secretions (cervical secretions),and all received multi-antibiotics therapies for several times. The patients were randomly divided into two groups: TCM group with 35 patients (aging 25-45) who had an average age of 32.05±8.73, and the black control group with 16 patients (aging 22-45) who stopped using drugs and were of an average age of 31.20±9.00.All patients of the two groups had no severe systemic diseases.

Experimental methods

Materials: Internal secretions from cervical canals(urethras) of the patients were taken as samples by bacteria-free long cotton swabs. And they were cultured for 48-72h with mycoplasma culture identification/counting/drug sensitivity kits produced by Haitai Biological Pharmaceutical Co., Ltd. in Zhuhai Special Economic Zone of China. There were 4 categories(10 species) of antibiotics in this kit: tetracyclines(minocycline and doxycycline), macrolides(azithromycin, roxithromycin, clarithromycin and josamycin), quinolones (sparfloxacin, ofloxacin and ciprofloxacin), and aminoglycosides (spectinomycin).Methods: The two groups of patients stopped taking antibiotics and sexual activities. Patients of TCM group were given oral administration of Cangbai Shiduqing Granule. The ingredients of Cangbai Shiduqing Granule were: ChaoCangZhu (RhizomaAtractylodis)10g, HuangBai (Cortex PhellodendriChinensis)10 g, ShengHuangQi (Radix Astragali)10g, DanShen(Radix SalviaeMiltiorrhizae) 15g, BaiBianDou (Semen Lablab Album) 10g, ShengYiRen (Semen Coicis)30g, TuFuLing (RhizomaSmilacisGlabrae) 15g,KuShen(Radix SophoraeFlavescentis)10 g, MaChiXian(HerbaPortulacae)15g, MaBianCao (HerbaVerbenae)10g, HuZhang (Rhiaoma et Radix polygoniCuspidati)10g, BaiJiangCao (HerbaPatriniae) 15g, BaiTouWeng(Radix Pulsatillae) 10g, and BaiHuaSheSheCao（HerbaHedyotis）15g (KangRenTang Pharmacy Limited Company). Cangbai Shiduqing Granule was dissolved in 200ml warm water and taken orally for 2 times each day, and its course for treatment was of 4 weeks. The patients of this group were reexamined after the medicine was stopped 1 week later. Patients of the black control group were not given any medicine, only provided with health education, such as to stop vaginal irrigation, live a regular life, go to bed early and get up early, regulate emotion, eat bland but nutritive food and etc. The patients of this group were reexamined after 4 weeks. When reexamined, both groups of patients were required to provide cervical (urethral) secretions for mycoplasma cultivation and drug sensitive tests(the reexamine time for patients with menstruation was delayed to the end of menstruation). The culture and drug sensitive results were examined to observe the sensitivity transformation situation of drug-resistant antibiotics.

Statistical analysis

The results were analyzed using SPSS19.0.Measurement data were indicated as “mean value± standard deviation” (x±s). T-test was used to analyze measurements which conformed with normal distributions and non-parametric test was applied to analyze measurements which conformed with nonnormal distributions. Chi-square test was used to analyze enumeration data. There were statistical differences (P＜0.05), and there were significant statistical differences (P＜0.01).

RESULTS

After 4 weeks of oral administration, 35 patients of TCM group felt that the symptoms disappeared or were alleviated. 22.86% (8/35) of the cultured mycoplasma showed negative while 77.14% showed positive.81.48% (22/27) of the positive cases showed varying degrees of sensitive reactions to the above 4 categories of antibiotics, and the remaining 18.52% (5/27) showed no sensitive reactions to antibiotics. As to 16 patients of the black control group, 100.00% (16/16) of the cultured mycoplasma showed positive. 31.25% (5/16) of the positive cases showed varying degrees of sensitive reactions to the above 4 categories of antibiotics, and the remaining 67.75% (11/16) showed no change in multidrug resistance. Comparative results were showed in the following table.

The table showed comparative results (%) of drug sensitive tests of the 2 groups after the treatment was over 1week later.

The above results showed that: after the treatment of 4 weeks, the sensitivity of mycoplasma strains to antibiotics of TCM group was obviously higher than that of the control group, especially for the categories of tetracyclines and macrolides. The sensitive reaction to quinolones was slightly poorer, but still 50% drugresistant strains transformed to sensitive strains. The drug sensitivity of the control group had certain degree of change, but the cases of sensitivity improvement were ＜20%, which wasobviously lower than that of TCM group, withP＜0.01.

DISCUSSIONS

UU has no cell wall, so it has natural drug resistance to antibiotics that can influence the synthesis of cell walls. In clinical treatment, antibiotics used in ribosome are the priorities to choose, mainly including tetracyclines, macrolides and quinolones. However,the drug resistance of UU is increasingly stronger due to factors like repeated infection, repeated deferment,abuse of antibiotics and substandard treatments. Many materials show1,2that UU has strong drug resistance to three categories of medicine. Therefore, to solve the problem of UU drug resistance is of great significance.In light of this problem, the methods such as combined use of antibiotics, incremental dose and intramuscular injection or intravenous drip instead of oral administration, are often adopted to improve the clinical efficacy, but the adverse reaction rate rises markedly3.The adverse reactions include: (1)increasing of liver and kidney damage as well as toxic and side effects,such as auditory nerve damage, development disorder of tooth enamel, inhibited bone growth, kidney failure,hematopoietic dysfunction and other severe damages to health; (2) disturbance of normal microf lora balance,internal environment disturbance and conditioned pathogen infection, such as double infections caused by fungus or virus; (3) induced generation of new drugresistant strains, even super drug-resistant strains, which makes the choice of medicine more difficult.

Now many studies show that the treatment of integrated Chinese traditional and western medicine can increase the sensitivity of drug-resistant antibiotics4,5,but they are limited to the observation of clinical effect,

bacterio stasis in vitro experiment or other aspects, the reversion mechanism of TCM to antibiotic resistance has not been clear and related literatures on TCM reversion on UU antibiotic resistance are rare. Hence,we made preliminary observations of the effective cases of TCM increasing the sensitivity of drug-resistant antibiotics, which will lay a foundation for the further study on TCM reversion action on the drug resistance of antibiotics.

Cangbai Shiduqing Granule takes BaiTouWeng (Radix Pulsatillae), KuShen (Radix SophoraeFlavescentis),BaiHua SheSheCao (Herba Hedyotis), MaChiXian(HerbaPortulacae), TuFuLing (Rhizoma Smilacis Glabrae) to clear away heat and toxic materials,promote diuresis and treat strangury; it takes CangZhu(Rhizoma Atractylodis), HuangBai (Cortex Phellodendri Chinensis), HuangQi (Radix Astragali), BaiBianDou(Semen Lablab Album), ShengYiRen (Semen Coicis)to reinforce qi, strengthen spleen, eliminate internal dampness and relieve internal heat or fever; and it takes DanShen (Radix SalviaeMiltiorrhizae), HuZhang(Rhiaoma et Radix polygoniCuspidati), MaBianCao(HerbaVerbenae), BaiJiangCao (HerbaPatriniae) to invigorate the circulation of blood, remove blood stasis,eliminate internal dampness and relive pains. These ingredients consider both purgation and tonification,strengthening body resistance and eliminating evil-qi;primarily, they clears away heat and toxic materials,so as to purge the remaining evils; secondarily, they strengthen spleen to tonify the source of acquired constitution. Thus, both deficiency and excess are treated. Modern pharmacological research shows that saponins in TuFuLing (RhizomaSmilacisGlabrae) and the berberines and amurensin in HuangBai (Cortex Phellodendri Chinensis) have antisepsis and antiinflammation effects. Saponins of astragali radix can significantly increase the total white blood cells in blood, and promote the phagocyte function and bactericidal ability of neutrophile granulocytes and macrophagocytes. Astragalus polysaccharides can stimulate the reproduction of NK cells and induce the production of interferon, etc. BaiJiangCao(HerbaPatriniae) has anti-inflammatory effects, which can improve the immune function and promote tissue repair.According to the research results, we can see that Cangbai Shiduqing Granule6-11has direct resistance effect on UU, and can also increase the sensitivity of pathogenic bacteria to antibiotics. In this study, the sensitivity of pathogenic bacteria to antibiotics for some patients in the control group also got certain degrees of change through themselves’ regulation and immunity recovery. But during the same period, the inactivation degree to pathogenic bacteria and the sensitivity change degree of pathogenic bacteria to antibiotics were of significant difference (P＜0.01) between the two groups. Our research results show that, the Chinese herbal compound prescription of Cangbai Shiduqing Granule has reversion action on the drug resistance of antibiotics, but the mechanism has not been clarified and needs further study.

1 Liu WJ, Wang GW, Xie LC, et al. Female nongonococcal urethritis of mycoplasma and choamydiae infections and drug susceptivity analysis. Hainan Medical J, 2012,23(2):99-101.

2 Chen HW, Zhang YM, Qiu SJ. Mycoplasma infection and srug susceptivity in qutpatients with nongonococcal Urethritis. Jilin Medical J, 2012, 33 (1):72-73.

3 Li J, Lin Y. clinical observation of three methods in treatment of male nongonococcal urethritis. China J Leprosy and Skin Diseases, 2004, 20(4):339.

4 Zheng YP, Yang N. Combination of TCM and Western medicine in treatment of nongonococcal urethritis. Jiangxi J Tradit Chin Med, 2004, 35(257):44.

5 Hong HY, Zheng LJ, Huang WP. Combination of clindamycin and TCM in treatment of 63 cases of reproductive tract infections of mycoplasma and chlamydia.Fujian J Tradit Chin Med, 2003, 34(5):30-31.

6 Cai LL, Li YW, Zhang FC. Study on the Cangbai Shiduqing granule making azithromycin-resistant UU strain sensitive to azithromycin and the relationship with blood IL-2, IL-12.China J Tradit Chin Med and Pharmacy, 2012, 27(1):172-175.

7 Mei Q, Li YW, Li N, Zhang XJ. Effect of Chinese medicine CangBaiShiDuQing on interleukin-12 concentration of murine vaginal mucosa. The Chin J Human Sexuality, 2013,22(12):58-62.

8 Li N, Li YW, Mei Q, et al. Intervention of Cang Bai Shi Du Qing on ureaplasma urealyticum model in female mice. The Chinese J Human Sexuality, 2013, 22(2):57-58.

9 Li YW, Sun ZX. Observation on the therapeutic effect of Cangbai Shiduqing granule in treating the patients with urethal and cervical tract infection by ureaplasma urealyticum. The Chinese J Human Sexuality, 2008,17(5):7-10.

10 Sun YM, Li YW, Zhang YC, et al. Establishment of ureaplasma urealyticum model in female mice and rats. The Chin J Human Sexuality, 2012, 21(3):24-25.

11 Li YW, Sun ZX, Zhang FC, et al. An experimental study on the effect of chinese durg CangBaiShiDuQing on the inhibition of ureaplasma urealyticum. The Chin J Human Sexuality, 2008, 17(10):10-17.

Funding:1. National Natural Science Fund（No:81072811);2. Beijing Administration of Traditional Chinese Medicine Development Plan in 13th Five-Year of Key Specialty of Dermatology; 3. Project from Human Resources Dept of Dongfang Hospital “Dongfang Famous Doctors”

*Corresponding author:Email:lingling89159166@126.com

(Accepted: March 12, 2017)