Quantitative analysis of hormones and inflammatory cytokines in Chlamydia trachomatis-infected women with tubal ectopic pregnancy and early intrauterine pregnancy

In this data, non-pregnant women during the menstrual cycle, women with normal intrauterine pregnancy (IUP), and women with tubal ectopic pregnancy (EP) after informed consent were included. The serum levels of 17β-estradiol, progesterone, testosterone, beta-human chorionic gonadotropin, interleukin (IL)-1β, IL-4, IL-6, IL-7, IL-8, IL-10, tumor necrosis factor α (TNFα), and interferon-γ (IFN-γ), epidermal growth factor, the Chlamydia (C.) trachomatis IgG and HSP60 were analyzed. Receiver operating characteristic analysis was used to assess the diagnostic discrimination of tubal EP and gestational age-matched IUP. Our data show that C. trachomatis infection is associated with IL-8 levels, which had excellent discriminative validity in positively identifying tubal EP (concomitant with C. trachomatis infection) from IUP and non-pregnant conditions regardless of C. trachomatis infection.


a b s t r a c t
In this data, non-pregnant women during the menstrual cycle, women with normal intrauterine pregnancy (IUP), and women with tubal ectopic pregnancy (EP) after informed consent were included. The serum levels of 17β-estradiol, progesterone, testosterone, beta-human chorionic gonadotropin, interleukin (IL)-1β, IL-4, IL-6, IL-7, IL-8, IL-10, tumor necrosis factor α (TNFα), and interferon-γ (IFN-γ), epidermal growth factor, the Chlamydia (C.) trachomatis IgG and HSP60 were analyzed. Receiver operating characteristic analysis was used to assess the diagnostic discrimination of tubal EP and gestational age-matched IUP. Our data show that C. trachomatis infection is associated with IL-8 levels, which had excellent discriminative validity in positively identifying tubal EP (concomitant with C. trachomatis infection) from IUP and non-pregnant conditions regardless of C. trachomatis infection.
& Value of the data 1. The data indicate that using a combination of serum IL-8 and steroid hormone levels can possibly differentiate a certain group of women with tubal EP from those with normal early IUP. 2. The data are useful for dissecting the molecular mechanism of different cytokines in the Fallopian tube after C. trachomatis infection and for how they participate in the development of tubal EP. 3. The data may provide a new diagnostic opportunity for tubal EP in women with C. trachomatis infection.
1. Data 1. The demographics and laboratory characteristics of the normal menstrual cycle women and those with IUP and EP are shown in Table 1. The rates of C. trachomatis infection in the women with tubal EP, IUP, and no pregnancy were 86.11%, 60.00%, and 55.40%, respectively (Table 2). 2. Although there was no significant difference in IL-8 levels between IUP and non-pregnancy, IL-8 levels were significantly higher in C. trachomatis-positive women with tubal EP than in women with IUP and in non-pregnant women regardless of C. trachomatis infection status ( Table 2). The diagnostic accuracy of the various parameters (E2, P4, T levels and the E2:P4, E2:EGF, E2:IL-8, P4:    EGF, and P4:IL-8 ratio) in the tubal EP and early IUP groups was evaluated by ROC analysis (Fig. 1). The ROC analysis showed that the IL-8 level had excellent discriminative validity in positively identifying tubal EP (concomitant with C. trachomatis infection) from IUP and non-pregnant conditions regardless of C. trachomatis infection (Fig. 1). 3. An increase in IL-1β levels and a decrease in IL-10 levels were observed in C. trachomatis-positive women with tubal EP compared to C. trachomatis-positive women with IUP and C. trachomatispositive non-pregnant women ( Table 2). 4. There were no significant differences in IL-4, IL-6, IL-7, IL-8, TNFα, or IFN-γ levels among C.

Source of the Curve
trachomatis-positive or negative women under pregnant and non-pregnant conditions ( Table 2). 5. We also found that individual measurements of serum EGF levels were strongly related to early pregnancy outcomes for women with tubal EP and IUP ( Table 2).

Ethics statement
This study was approved by the Ethics Committees of the Obstetrics and Gynecology Hospital and Shanghai Medical College, Fudan University, China. All participants provided informed consent.

Experimental design and sample collection
All participants underwent clinical examination at the Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China. Clinical work-up included menstrual history as well as current cycle length and menstrual regularity. Exclusion criteria included use of estrogen-or progestincontaining medication within three months of the study, past EP, any gynecological pathology (e.g., endometriosis, fibroids, or any operation to the gynecological organs), infection, and smoking. All the patients whose pregnancies resulted from assisted reproductive technologies were also excluded from the study. Blood samples were collected into Sarstedt evacuated tubes without anticoagulant. All blood samples were centrifuged at 1000 Â g for 15 min, and the serum was stored at -80°C until batch analyses. The present study included non-pregnant and pregnant women (total n ¼225) who were subdivided into the following groups: 1. Group 1. The different stages of the menstrual cycle in non-pregnant women (n ¼139) were studied. Blood samples were collected at the scheduled visits during their menstrual cycle. Menstrual cycle day was established using the criteria reported by Noyes et al. [1]. Sample dating Table 3 The intra-assay and inter-assay % CV for the human cytokine magnetic bead kit and angiogenesis/growth factor magnetic bead panel 1 kit.

Main outcome measures
All sera were stored at -80°C before performing the assays, and aliquots that had not been previously thawed were used in the present study. Samples were tested in duplicate and analyzed individually. Radioimmunoassays (RIA) were performed at Beijing Free Co. (China), and enzymelinked immunoassays (ELISA) were performed at the Department of Integrative Medicine and Neurobiology, Shanghai Medical College, Fudan University (China). The averages of the duplicate readings for each standard, control, and individual samples were used for the analyses. [2]. As opposed to accuracy, sensitivity and specificity are not dependent on the prevalence of the disease in the sample. Thus, ROC curve analysis provides a description of disease detectability that is independent of both disease prevalence and decision threshold effects. For ROC analysis, women with EP were considered affected, and IUP as unaffected. ROC curves were constructed by plotting the sensitivity (true-positive) on the ordinate as a function of the complement of specificity (falsepositive) for all possible cut-off values of the diagnostic test [3]. Greater deviation toward the left upper corner of the curve indicates better detection of tubal EP.