Pharmacokinetics of cationic liposome-encapsulated doxycycline in mice challenged with genital infection by Chlamydia trachomatis

Our previous studies demonstrated the efficacy of cationic liposome-encapsulated doxycycline (CaL-DOX) on the course of infection with Chlamydia trachomatis in vitro and in vivo. In this investigation, the pharmacokinetics of CaL-DOX are reported. Female mice inoculated intravaginally with 4.84 x 10(5) inclusion-forming units were treated intramuscularly, 3 days postinfection, as determined by a preliminary study, with 0.1 ml of unencapsulated doxycycline (DOX) or CaL-DOX, at 10 microg/ml body weight for 3 consecutive days. Sampling was performed at 12, 24, 48, 72 and 96 h after treatment. The microbiological test was used to measure the DOX concentration in sera, liver, kidneys and genital organs. The maximum concentration in kidneys, liver and genital organs was higher in mice treated with DOX than in those treated with CaL-DOX. However, in sera, the amount was similar to that in mice treated with DOX. The DOX concentration found in mice treated with CaL-DOX was much less than in those treated with unencapsulated DOX. This may have a direct impact on the secondary effects caused by the medication. Decreased secondary effects should have a positive outcome on patient physical and mental health. Even if this study is the only one of its kind so far, CaL-DOX could be an alternative solution for the treatment of chlamydial infections.     

Characteristics of murine model of genital infection with Chlamydia trachomatis and effects of therapy with tetracyclines, amoxicillin-clavulanic acid, or azithromycin.

Following intravaginal inoculation of progesterone-treated outbred mice with Chlamydia trachomatis MoPn, 4 to 6 log10 inclusion-forming units were recovered in vaginal swabs for 21 days but all animals were culture negative after 28 days. Serum antibody titers were elevated and remained high for at least 70 days. Between 28 and 70 days, upper tract infection (inflammation and distension of the uterine horns, occlusion of oviducts with inflammatory exudate, pyosalpinx, and hydrosalpinx) was seen in > 80% of the animals. Mice were dosed orally, commencing at 7 days after infection, with minocycline, doxycycline, or amoxicillin-clavulanate. Further groups received azithromycin either as a single high dose or as lower once-daily doses. In addition, minocycline and amoxicillin-clavulanate were administered at 24 h after infection, and this early treatment prevented elevation of antibody titers whereas delayed therapy did not. Vaginal swabs from mice in all treatment regimens were culture negative except for 25% of mice receiving either early amoxicillin-clavulanate or low-dose azithromycin, which yielded low numbers (20 to 70 inclusion-forming units) of chlamydiae. Numbers of fertile mice in the early treatment regimens and their litter sizes were similar to those of noninfected controls, although 25% of amoxicillin-clavulanate-treated mice had unilateral hydrosalpinges. In comparison, 88% of untreated mice developed hydrosalpinges and only 25% conceived. Delayed dosing did not affect the outcome of amoxicillin-clavulanate therapy but did diminish the protective efficacy of minocycline such that 50% of treated mice had either unilateral hydrosalpinges or ovarian abscesses. Doxycycline and azithromycin were highly effective in restoring fertility. This model makes possible the study of both short- and long-term outcomes of chlamydial infection.     

Ofloxacin versus doxycycline for treatment of cervical infection with Chlamydia trachomatis.

Women with culture-proven Chlamydia trachomatis cervical infection were randomized to receive either ofloxacin (300 mg) or doxycycline (100 mg), orally twice daily for 7 days. All 56 had negative cultures 5 to 9 days after treatment. Four weeks after treatment, 26 (93%) of 28 ofloxacin-treated patients and all 22 doxycycline-treated patients were cured. We conclude that 300 mg of ofloxacin given twice daily for 7 days provides effective therapy for chlamydial infection of the cervix.     

女性生殖道感染病原学检查及结果分析

目的 了解女性生殖道感染病原体感染状况,探讨HIV感染与相关病原体的关系。方法 采用高超倍镜检测阴道/宫颈分泌物及抽取静脉血分别进行相关病原体实验室检查。结果 门诊患者生殖道炎症为82.63%（6939/8398）,霉菌感染18.83%、滴虫感染3.57%、淋球菌感染1.45%、衣原体/支原体感染21.89%、细菌性感染28.6%、梅毒感染2.26%、HIV0.08%,38.51%的人有两种或两种以上病原体感染,其中合并HIV感染7例;健康体检组生殖道炎症19.93%（1656/8309）,霉菌感染7.7%、滴虫感染0.3%、淋球菌感染0.05%、衣原体/支原体感染2.0%、细菌性感染9.3%,4.28%的人有两种或两种以上病原体感染。结论 阴道炎患者以细菌性感染为主,宫颈炎者以衣原体/支原体感染为主,细菌性阴道炎患者是HIV感染的重要危险因子;超高倍镜能快速、准确检测多种病原体,是筛查生殖道常见病原体感染较理想的方法。     

Chlamydia and other sexually transmitted bacterial infections

Chlamydia trachomatis is the most common sexually transmitted bacterium worldwide. In Western Europe, the prevalence of gonorrhoea has decreased by more than 95% since the 1970ies; "tripper" and syphilis are essentially confined to high-risk groups while genital chlamydial infections affect people of all social classes, but information about chlamydia is still scarce in many European countries. Clinically genital chlamydial infections resemble gonorrhoea (dysuria, discharge, irregular bleeding, dyspareunia, perihepatitis) and may be mistaken for appendicitis. However, Chlamydia trachomatis persists longer and more often asymptomatic than Neisseria gonorrhoeae in the urogenital tract of men and women. About 20% of all chlamydia infected women suffer from partial or complete tubal occlusion. Chlamydia trachomatis is the leading cause of female infertility, but most of these women never experienced any clinical sign of pelvic inflammatory disease. Since particle concentrations are often very low in urine and cervical secretions only DNA-amplification tests, e.g. PCR or LCR, exhibit sufficient sensitivity for direct detection Chlamydia trachomatis. While Neisseria gonorrhoeae is eradicated by single-shot treatment with commonly used antibiotics like penicillins or cephalosporins Chlamydia trachomatis affords treatment for at least 10 days with doxycyline or macrolides. Partner treatment is essential to avoid reinfections. Condoms not only protect against HIV, but also against chlamydia, gonorrhoea and syphilis.     

Roxithromycin treatment of mouse chlamydial salpingitis and protective effect on fertility.

We used a mouse model of acute chlamydial salpingitis to evaluate the efficacy of roxithromycin in preventing irreversible inflammatory damage leading to tubal infertility. Female C3H/He mice were genitally inoculated with a human strain of Chlamydia trachomatis and then treated with roxithromycin glutamate subcutaneously. Treatment was initiated either 7 or 10 days postinfection (p.i.) and continued for 7 days at a dosage of 50 or 100 mg/kg of body weight per 24 h. The course of the disease was monitored serologically, bacteriologically, and histologically. At the end of the treatment, the mice were encaged with males and their reproductive capacity was recorded over a 19-week period. The protective effect of roxithromycin was assessed in terms of fertility parameters in comparison with values for noninfected control mice. When treatment was initiated on day 7 p.i. and given in twice-daily 25-mg/kg doses, all the mice remained fertile and the total number of offspring was similar to that of sham-infected mice (17.3 +/- 3.3 versus 17.2 +/- 2.3). When treatment was initiated on day 10 p.i. and given in a single daily dose of 50 or 100 mg/kg, 90 and 70% of the mice, respectively, remained fertile; however, in terms of total offspring, fertility was lower in the group treated with the lower dose (5.6 +/- 1.4 versus 13.0 +/- 3.8). Roxithromycin was found to be effective against C. trachomatis in the mouse genital tract, but fertility was only partially preserved when the time between infection and treatment was prolonged.     

Susceptibilities of Chlamydia trachomatis isolates causing uncomplicated female genital tract infections and pelvic inflammatory disease.

The in vitro susceptibilities of 45 recent clinical isolates of Chlamydia trachomatis obtained from women with asymptomatic genital tract infection, mucopurulent cervicitis, or pelvic inflammatory disease to doxycycline, azithromycin, ofloxacin, and clindamycin were determined. In addition, susceptibilities of 12 isolates to amoxicillin and trimethoprim-sulfamethoxazole were also determined. Isolates also were serotyped with a panel of monoclonal antibodies specific for chlamydial major outer membrane protein; 24 of 45 (53%) belonged to serovars Ia and E. For all isolates, the MIC range of doxycycline was 0.008 to 0.06 micrograms/ml, for trimethoprim-sulfamethoxazole it was 0.03 to 0.25 micrograms/ml, for azithromycin it was 0.125 to 2.0 micrograms/ml, for ofloxacin it was 0.5 to 1.0 micrograms/ml, for clindamycin it was 0.25 to 2.0 micrograms/ml, and for amoxicillin it was 0.25 to 4.0 microgram/ml. The ranges of minimum chlamydiacidal concentrations were generally 1 to 4 dilutions above the MICs of most agents, with a rank order similar to those of the MICs. Comparing the minimum chlamydiacidal concentrations for 90% of isolates tested, isolates causing asymptomatic infection belonged to a greater variety of serovars and were relatively more susceptible to doxycycline and azithromycin than isolates causing mucopurulent cervicitis or pelvic inflammatory disease; these differences in susceptibility were not detected among the other study agents. These data indicate that additional studies are needed to better define the apparent association of certain chlamydial serovars with the clinical severity of disease and the in vitro susceptibilities to certain antimicrobial agents.     

Intra-articular chlamydial antigen and inflammatory arthritis

Joint material from 133 patients with well-characterized inflammatory arthritis, including individuals likely to have suffered reactive arthritis, was studied. The majority of patients were also examined for the presence of genital tract infection with Chlamydia trachomatis. Fluorescein-conjugated monoclonal antibodies demonstrated the presence of C. trachomatis antigen in synovial fluid cell deposits or synovial sections from inflamed knee joints of seven patients with reactive arthritis. The significance of these findings is discussed, as is the low rate of detection of chlamydial antigen in either the genital tract or the joint from patients in this study. We emphasize the need for further work aimed at identifying the relevant immunogenic chlamydial antigens responsible for the initiation of reactive arthritis.     

Differential Susceptibilities to Azithromycin Treatment of Chlamydial Infection in the Gastrointestinal Tract and Cervix

Evidence from animal studies suggests that chlamydiae may persist in the gastrointestinal tract (GI) and be a reservoir for reinfection of the genital tract. We hypothesize that there may be a differential susceptibility of organisms in the GI and genital tracts. To determine the effect of azithromycin on persistent chlamydial gut infection, C57BL/6 and BALB/c mice were infected orally and genitally and treated with azithromycin (Az) orally (20, 40, or 80 mg/kg of body weight), and the numbers of chlamydiae were determined from cervix and cecal tissues. The Az concentration in the cecum and cervix was measured by high-performance liquid chromatography with electrochemical detection (HPLC-ECD). Az treatment cleared genital infection in both C57BL/6 and BALB/c mice; however, GI infection was not cleared with the same doses. HPLC data showed the presence of Az at both sites of infection, and significant amounts of Az were measured in treatment groups. However, no significant difference in Az levels between the cecum and the cervix was observed, indicating similar levels of Az reaching both sites of infection. These data indicate that antibiotic levels that are sufficient to cure genital infection are ineffectual against GI infection. The results suggest a reevaluation of antibiotic therapy for chlamydial infection.     

Diagnosis and treatment of Chlamydia trachomatis infection

Chlamydia trachomatis infection most commonly affects the urogenital tract. In men, the infection usually is symptomatic, with dysuria and a discharge from the penis. Untreated chlamydial infection in men can spread to the epididymis. Most women with chlamydial infection have minimal or no symptoms, but some develop pelvic inflammatory disease. Chlamydial infection in newborns can cause ophthalmia neonatorum. Chlamydial pneumonia can occur at one to three months of age, manifesting as a protracted onset of staccato cough, usually without wheezing or fever. Treatment options for uncomplicated urogenital infections include a single 1-g dose of azithromycin orally, or doxycycline at a dosage of 100 mg orally twice per day for seven days. The recommended treatment during pregnancy is erythromycin base or amoxicillin. The Centers for Disease Control and Prevention and the U.S. Preventive Services Task Force recommend screening for chlamydial infection in women at increased risk of infection and in all women younger than 25 years.     

Comparison of spiramycin and doxycycline for treatment of Chlamydia trachomatis genital infections.

We performed a single blind controlled multicenter study in which we compared the efficacy and safety of 100 mg of doxycycline versus those of 1 g (3 x 10(6) IU) of spiramycin given orally twice daily for 14 days in the treatment of culture-positive Chlamydia trachomatis genitourinary tract infections. A total of 367 patients were enrolled in the study, and 364 patients were evaluable for safety and 265 patients were evaluable for efficacy. The cure rate between treatment groups was not statistically significant, being 98% (125 of 128 patients) in the spiramycin group and 100% (133 of 133 patients) in the doxycycline group. Female patients who received spiramycin were more likely to report dysethesias that resolved after the completion of therapy. The results of the study show that spiramycin is an effective drug for the treatment of C. trachomatis infection and warrants further assessment over a shorter treatment period (7 days) and during pregnancy.     

Recommendations and rationale for the treatment of pelvic inflammatory disease

Pelvic inflammatory disease (PID) is one of the most common serious infections of nonpregnant women of reproductive age. Management of PID is directed at containment of infection. Goals of therapy include the resolution of clinical symptoms and signs, the eradication of pathogens from the genital tract and the prevention of sequelae including infertility, ectopic pregnancy and chronic pelvic pain. The choice of an antibiotic regimen used to treat PID relies upon the appreciation of the polymicrobial etiology of this ascending infection including Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium and other lower genital tract endogenous anaerobic and facultative bacteria, many of which are associated with bacterial vaginosis. Currently available evidence and the CDC treatment recommendations support the use of broad-spectrum antibiotic regimens that adequately cover the above named microorganisms. The outpatient treatment of mild-to-moderate PID should include tolerated antibiotic regimens consisting of an extended-spectrum cephalosporin in conjunction with either azithromycin or doxycycline. Clinically severe PID should prompt hospitalization and imaging to rule out a tubo-ovarian abscess. Parenteral broad-spectrum antibiotic therapy with activity against a polymicrobial flora, particularly Gram-negative aerobes and anaerobes, should be implemented.     

性传播疾病感染病原体检测分析

目的了解本地区性传播疾病（STD）中几种病原体的感染及药敏情况。方法淋球菌采用桂梅培养基培养法,沙眼衣原体采用单克隆抗体快速免疫法,支原体应用Mycoplasma IST试剂盒进行培养及药敏试验。结果887例检测标本中,病原体检测阳性者349例,检出率39．3％,其中单种病原体阳性者246例,占总阳性者的70．5％：混合阳性者103例,占29．5％。共检出各种病原体456株,药敏支原体较敏感药物有原始霉素、交沙霉素（Jos）、强力霉素（Dox）和氧氟沙星,分别为100％、94．4％、83．5％和73．9％。淋球菌（NG）对壮观霉素32例菌株全部敏感。结论解脲支原体、沙眼衣原体和淋球菌是性传播疾病的主要病原体,且混合感染不能忽视。对STD患者应尽可能做多种病原体检测和治疗。支原体和NG感染可旨选Jos、Dox和壮观霉索治疗。     

Randomized comparison of ofloxacin and doxycycline for chlamydia and ureaplasma urethritis and cervicitis

Fifty-eight males and 34 females with nongonococcal urethritis and/or cervicitis were treated to compare the efficacy and safety of 7-day regimens of oral ofloxacin 300 mg twice daily and doxycycline hyclate 100 mg twice daily. Forty-seven patients were randomized to receive ofloxacin and 45 patients to receive doxycycline. The microbiologic response rate was 97% (32/33) for both ofloxacin and doxycycline; the combined microbiologic and clinical cure rates were 98% for both treatment groups (ofloxacin 46/47, doxycycline 44/45). Ofloxacin was as effective as doxycycline in the treatment of chlamydial infections (96% vs. 100%). In patients with Ureaplasma urealyticum, the initial response was complete with either drug, but recurrence of infection was observed with both treatment groups (1 of 4 patients in the ofloxacin group and 2 of 11 patients in the doxycycline group). In the treatment of mixed Chlamydia trachomatis and U. urealyticum infections, all 5 patients treated with ofloxacin and 3 of 4 patients treated with doxycycline were cured. In symptomatic patients whose initial cultures were negative, clinical cures were complete with both drugs, but Ureaplasma was isolated at 3 or more weeks post-treatment in 2 patients treated with ofloxacin. In a study of single-dose ofloxacin treatment of uncomplicated gonorrhea, Neisseria gonorrhoeae was eradicated in all subjects, but C. trachomatis was not reliably eradicated. Both drugs were well tolerated with only minimal adverse effects reported in either treatment group. A multiple-dose regimen of ofloxacin appears to be a highly effective and well-tolerated alternative to doxycycline in nongonococcal sexually transmitted disease.     

Abdominal pain. Chlamydia as culprit

Chlamydia trachomatis infection in the lower part of the genital tract of young girls and women may ascend to produce endometritis, salpingitis, perihepatitis, and other localized or generalized abdominal diseases. The resultant pain syndromes mimic a number of other common conditions that must be differentiated. A careful history and physical examination, with attention to historical and physical evidence of sexually transmitted disease, will alert the clinician to the possibility of chlamydial infection. Laboratory tests for C trachomatis may be helpful. However, tests of specimens from the lower genitourinary tract may yield negative results in patients with disease of the upper part of the genital tract and abdomen. Prompt recognition and treatment not only alleviate pain but also may help prevent inflammatory sequelae such as chronic painful adhesions, small-bowel obstruction, and tubal infertility. Costly workups and unnecessary surgery may also be avoided.     

Efficacy of erythromycin in the treatment of inner city pregnant women with cervical Chlamydia trachomatis infection.

In this retrospective study, the efficacy of screening for and treating cervical Chlamydia trachomatis infection was evaluated in a pregnant population at increased risk for chlamydial infection. Over a 2 1/2-year period, 5.75% (338) of the 5,875 women tested were found to be infected with this organism. Of the 323 women patients available for follow-up, 76% (244) were successfully treated and 24% (79) remained infected throughout their pregnancies. Forty (12%) patients became infected during pregnancy, while 26 (8%) were reinfected during pregnancy, despite treatment with erythromycin. Twenty-seven (8%) patients had their first antenatal visit and cervical swab less than a week before delivery. The gestational age at which the first cervical chlamydial swab was obtained was significantly more advanced in patients who remained infected (30.23 +/- 6.2 weeks) than those who were successfully treated (22.15 +/- 7.66 weeks; P = 0.00001). The data suggest that in a pregnant population considered to be at increased risk for C trachomatis infection: (1) there is a subgroup of patients with a high risk of remaining infected or becoming reinfected with C trachomatis during pregnancy despite treatment with erythromycin and (2) repeated prenatal testing and treatment of those infected is necessary to detect and eradicate maternal chlamydial infection.     

2018－2019年浙江省生殖道沙眼衣原体感染重复报告情况调查

目的了解浙江省2018 — 2019年生殖道沙眼衣原体感染重复报告情况,为掌握疫情数据制定相应管理措施提供参考。方法收集中国疾病预防控制信息系统传染病监测信息系统2018 — 2019年浙江省上报的报告病例,进行个案查重,计算重复报告率,并进行统计学分析。结果2018年1月1日至2019年12月31日报告生殖道沙眼衣原体感染病例59 826例,重复报告率为1.29%。 地区重复率为0.28%～5.68%,差异有统计学意义（χ2＝300.970,P＜0.001）;各年龄组的重复报告率为0.67%～1.65%（χ2＝50.060,P＜0.001）;妇产（科）医院及妇幼保健院（所、站）重复报告率（1.54%）高于其他的医疗机构（1.19%）χ2＝10.790,P＜0.05）。结论浙江省生殖道沙眼衣原体感染存在重复报告的情况,应加强医疗机构和疾病预防控制机构查重工作。     

Aerosol delivery of liposome-encapsulated ciprofloxacin: aerosol characterization and efficacy against Francisella tularensis infection in mice.

The aerosol delivery of liposome-encapsulated ciprofloxacin by using 12 commercially available jet nebulizers was evaluated in this study. Aerosol particles containing liposome-encapsulated ciprofloxacin generated by the nebulizers were analyzed with a laser aerodynamic particle sizer. Mean mass aerodynamic diameters (MMADs) and geometric standard deviations (GSDs) were determined, and the drug contents of the sampling filters from each run onto which aerosolized liposome-encapsulated ciprofloxacin had been deposited were analyzed spectrophotometrically. The aerosol particles of liposome-encapsulated ciprofloxacin generated by these nebulizers ranged from 1.94 to 3.5 microm, with GSDs ranging from 1.51 to 1.84 microm. The drug contents of the sampling filters exposed for 1 min to aerosolized liposome-encapsulated ciprofloxacin range from 12.7 to 40.5 microg/ml (0.06 to 0.2 mg/filter). By using the nebulizer selected on the basis of most desirable MMADs, particle counts, and drug deposition, aerosolized liposome-encapsulated ciprofloxacin was used for the treatment of mice infected with 10 times the 50% lethal dose of Francisella tularensis. All mice treated with aerosolized liposome-encapsulated ciprofloxacin survived the infection, while all ciprofloxacin-treated or untreated control mice succumbed to the infection (P < 0.001). These results suggest that aerosol delivery of liposome-encapsulated ciprofloxacin to the lower respiratory tract is feasible and that it may provide an effective therapy for the treatment of respiratory tract infections.     

下生殖道性传播病原体分布特点及其与高级别宫颈鳞状上皮内病变的关系

目的 探讨女性下生殖道性传播病原体的分布特点及其与高级别宫颈鳞状上皮内病变(HSIL)的关系.方法 选取246名进行宫颈癌筛查的女性作为研究对象,对其进行下生殖道涂片病理学检查.筛查出82例下生殖道性传播病原体阳性患者,并对其进行宫颈组织细胞学检查.共有38例患者被诊断为HSIL阳性,将此部分双阳性患者设为阳性组,HS...     

Vaccination against Chlamydial Genital Tract Infection after Immunization with Dendritic Cells Pulsed Ex Vivo with Nonviable Chlamydiae

Chlamydia trachomatis, an obligate intracellular bacterial pathogen of mucosal surfaces, is a major cause of preventable blindness and sexually transmitted diseases for which vaccines are badly needed. Despite considerable effort, antichlamydial vaccines have proven to be elusive using conventional immunization strategies. We report the use of murine bone marrow–derived dendritic cells (DC) pulsed ex vivo with killed chlamydiae as a novel approach to vaccination against chlamydial infection. Our results show that DC efficiently phagocytose chlamydiae, secrete IL-12 p40, and present chlamydial antigen(s) to infection sensitized CD4⁺ T cells. Mice immunized intravenously with chlamydial-pulsed DC produce protective immunity against chlamydial infection of the female genital tract equal to that obtained after infection with live organisms. Immunized mice shed ∼3 logs fewer infectious chlamydiae and are protected from genital tract inflammatory and obstructive disease. Protective immunity is correlated with a chlamydial-specific Th1-biased response that closely mimics the immune response produced after chlamydial infection. Thus, ex vivo antigen-pulsed DC represent a powerful tool for the study of protective immunity to chlamydial mucosal infection and for the identification of chlamydial protective antigens through reconstitution experiments. Moreover, these findings might impact the design of vaccine strategies against other medically important sexually transmitted diseases for which vaccines are sought but which have proven difficult to develop.