Research on the mechanism of Chinese herbal medicine Radix Paeoniae Rubra in improving chronic pelvic inflammation disease by regulating PTGS2 in the arachidonic acid pathway.

Radix Paeoniae Rubra (RPR) is a traditional Chinese medicine with anti-inflammatory effects that has been used in chronic pelvic inflammation disease (CPID) therapy. However, research on the mechanism of RPR in CPID therapy is lacking. Here, we used a network pharmacology method to screen targets and found that the PTGS2 target in the arachidonic acid (AA) pathway was significantly related to CPID. Then, regarding the molecular mechanism, it was further confirmed that RPR may reduce the development of CPID by regulating the PTGS2 target. The CPID rat model was established by mixed bacterial infection. We verified the expression of PTGS2 by immunohistochemical analysis, western blotting assays to detect the expression of PTGS2 protein, and polymerase chain reaction detection of PTGS2 mRNA expression. It was observed that the PTGS2 target decreased significantly after RPR administration at different doses. It is suggested that RPR can reverse the abnormal expression of PTGS2 in CPID rats. We believe that RPR is effective in the treatment of CPID, and RPR can reduce the inflammatory symptoms of CPID by regulating the level of PTGS2 in the AA pathway.

A novel fluorescence method for activity assay and drug screening of T4 PNK by coupling rGO with ligase reaction.

T4 polynucleotide kinase (PNK) is the primary member of the 5'-kinase family that can transfer the γ-phosphate residue of ATP to the 5'-hydroxyl group of oligonucleotides. In this article, using the differential quenching ability of reduced graphene oxide (rGO) towards the fluorophore-labeled DNA probe, we propose a novel method for detecting T4 PNK activity assisted by ligase reaction. Under the optimized conditions, the detection limit of T4 PNK was estimated to be 0.0002 U µL-1 in the linear region of 0.001 U µL-1-0.1 U µL-1. Additionally, the developed method was used to screen regulators of T4 PNK from natural compounds. The compound f isolated from the root of Kadsura coccinea (Lem.) A.C. Smith was found to stimulate T4 PNK activity in a concentration-dependent manner in vitro. Finally, the method was used to monitor the relation of T4 PNK activity with pelvic inflammatory disease (PID). The results demonstrated that the development of this disease could inhibit T4 PNK activity to some extent. In summary, the above data indicate that the method not only provides a universal platform for monitoring T4 PNK activity, but also shows great potential to be used in drug screening and clinic diagnosis.

Evaluation of pelvic inflammatory disease potential in cholinesterase inhibitor pesticide-exposed females.

Cholinesterase inhibitor pesticides, mainly organophosphates and carbamates, are commonly used in Egypt. Chronic exposure of males and females working in agriculture is expected. The study aimed to relate exposure to cholinesterase inhibitor pesticides to the development of pelvic inflammatory disease (PID). This is a case-control study that was conducted among 84 females. Seventy patients complained of pelvic inflammatory disease visited the outpatient Gynecology and Obstetrics Clinic. Fourteen females were not suffering from PID and were chosen as a control group. Red blood cells' cholinesterase activity was measured in blood. Cervical swaps were collected, and cultures were submitted for microbiological examination. The results showed that cholinesterase activities were significantly depressed in exposed females (6.36 ± 0.8 µmoles/min/ml red cells) when compared to non-exposed (7.5 ± 1.2 µmoles/min/ml red cells), and both were significantly depressed when compared with healthy females (9.17 ± 0.7 µmoles/min/ml red cells). The correlation coefficient (r) between previous exposure and the laboratory confirmed cervical infection was 0.31, with a P value of 0.009. The study concluded that exposure to cholinesterase inhibitor pesticides could increase the occurrence of pelvic inflammatory disease.

Significant elevation and correlation of plasma neutrophil gelatinase associated lipocalin and its complex with matrix metalloproteinase-9 in patients with pelvic inflammatory disease.

BACKGROUNDS: To detect the expression of plasma neutrophil gelatinase associated lipocalin (NGAL) and its complex with matrix metalloproteinase-9 (MMP-9) in patients with pelvic inflammatory disease (PID). METHODS: Enzyme-linked immunosorbent assay was used to measure the levels of plasma NGAL and NGAL/MMP-9 complex. RESULTS: The levels of plasma NGAL or NGAL/MMP-9 complex were increased in patients with PID compared with those in normal controls and decreased significantly after treatment. Pre-treatment plasma level of NGAL was significantly correlated with WBC and neutrophil counts. In patients with PID, plasma level of NGAL/MMP-9 complex was correlated with plasma level of NGAL or MMP-9 significantly. In predicting PID, the sensitivities of NGAL and NGAL/MMP-9 complex were 76.6% and 78.1%; the negative predictive values, 72.7% and 74.5%. CONCLUSIONS: Plasma NGAL and NGAL/MMP-9 complex may act as diagnostic adjuvant biomarkers for PID. In patients with PID, about 80% have plasma levels of NGAL or NGAL/MMP-9 complex level >10.04 ng/ml or 2.33 ng/ml, respectively.

Markedly elevated plasma myeloperoxidase protein in patients with pelvic inflammatory disease who have A allele myeloperoxidase gene polymorphism.

OBJECTIVE: To compared the plasma levels and the genetic polymorphism of myeloperoxidase in patients with pelvic inflammatory disease (PID) and healthy controls. DESIGN: A random consecutive study. SETTING: University hospital. PATIENT(S): Forty-four women who were diagnosed with PID. INTERVENTION(S): Collected blood specimens of patients before and after they received treatment. MAIN OUTCOME MEASURE(S): ELISA and polymerase chain reaction-restriction fragment length polymorphism were respectively used to measure the plasma levels and genetic polymorphism of myeloperoxidase. RESULT(S): We found the plasma level of myeloperoxidase was elevated in PID patients compared with that in normal controls and decreased significantly compared with that in the same patients after they received treatment. PID patients with myeloperoxidase G/A alleles significantly tended to have elevated plasma concentration of myeloperoxidase, whereas PID patients with G/G homozygous alleles did not. CONCLUSION(S): Elevated expression of myeloperoxidase may be involved in the pathogenesis of PID and could be useful for the diagnosis of PID. Furthermore, G/A alleles of myeloperoxidase may contribute to such elevation in PID patients.

Increased activity of lysosomal enzymes in the peritoneal fluid of patients with gynecologic cancers and pelvic inflammatory disease.

PURPOSE: To investigate whether the activity of lysosomal enzymes is increased in the peritoneal fluid of patients with gynecologic cancers compared to activity in the peritoneal fluid from normal subjects and those with pelvic inflammatory disease, and fluid from benign ovarian cysts. PATIENTS AND METHODS: beta-glucuronidase, beta-galactosidase, and alpha-mannosidase activity was measured in the peritoneal fluid from patients with gynecologic cancer, pelvic inflammatory disease, and normal subjects, and fluid from benign ovarian cysts. RESULTS: The mean+/-SD of beta-glucuronidase, beta-galactosidase, and alpha-mannosidase activity in the gynecologic cancers was 120+/-50 nmol, 203+/-86 nmol, and 240+/-119 nmol 4-methylumbelliferone/ml/h, respectively; in the normal control subjects it was 22+/-9 nmol, 46+/-10 nmol, and 80+/-23 nmol, respectively (P=0.00003, 0.0001, and 0.0001, respectively). The activity was increased even in cases without malignant cells in the peritoneal fluid. In pelvic inflammatory disease it was 148+/-82 nmol, 278+/-112 nmol, and 291+/-140 nmol, respectively. The activity in the fluid of the ovarian cysts was similar to that of the normal peritoneal fluid. There was a significant positive correlation between enzyme activity and stage of cancer, that was stronger for beta-glucuronidase (r=0.889, P=0.003). CONCLUSION: The increased lysosomal enzyme activity in gynecologic cancers, without overlapping between patients and normal subjects or benign ovarian cyst fluid, indicates that such measurements might be applied for diagnostic purposes.

Maternal serum creatine kinase as a biochemical marker of tubal pregnancy.

OBJECTIVE: To evaluate the role of serum creatine kinase as a possible marker for diagnosis of tubal pregnancy. METHODS: Five groups of patients were included in this prospective study: group A, 20 patients with tubal pregnancies; group B, 20 patients with missed abortions; group C, 20 patients with pelvic inflammatory disease; group D, 10 patients with acute appendicitis; group E (controls), 20 patients with normal pregnancies matched for age and gestation. Total creatine kinase levels were measured in the serum of all five groups. RESULTS: Creatine kinase levels were found to be > or =75 IU/l in all patients with tubal pregnancies, which was significantly higher than in the other four groups (P < 0.0001). CONCLUSION: Our results indicate that maternal serum creatine kinase can be an important biochemical marker in suspected tubal pregnancy.

Endometrial phospholipases A2, polycystic ovaries and pelvic pain.

OBJECTIVE: To compare the activity of calcium independent phospholipase A2 in the endometrium of women with polycystic ovaries and in women with normal ovaries, and to investigate the influence of chronic pelvic pain on phospholipase A2 activity. DESIGN: A prospective descriptive study. SETTING: The Samaritan Hospital for Women and the Unit of Metabolic Medicine, St Mary's Hospital Medical School, London. SUBJECTS: 73 women attending the Samaritan Hospital for Women for clip sterilization, infertility, early recurrent miscarriage or pelvic venous congestion. 45 of these women had no pelvic pain, 22 had normal ovaries and 23 had polycystic ovaries diagnosed by ultrasound. The other 28 women had chronic pelvic pain, 14 of them had normal ovaries and 14 polycystic ovaries. INTERVENTIONS: Endometrial tissue was obtained at curettage or from the excised uterus in the proliferative or secretory phase of the menstrual cycle. The activities of calcium dependent (type 1) and calcium independent (type 2) phospholipase A2 isoenzymes were measured in all endometrial samples. RESULTS: In all the women phospholipase A2 type 1 activity, was significantly higher in the secretory phase than in the proliferative phase (P less than 0.001). There was no difference between women with normal ovaries and those with polycystic ovaries at either stage of the cycle irrespective of whether or not chronic pelvic venous congestion was present. In women with normal ovaries, both with and without chronic pelvic pain, phospholipase A2 type 2 activity was significantly higher in the secretory phase than in the proliferative phase (P less than 0.02 and P less than 0.05 respectively) but in women with polycystic ovaries, with and without chronic pelvic pain, there was no significant difference in activity between the two phases of the cycle. Women with polycystic ovaries had markedly higher proliferative phase type 2 activity than women with normal ovaries (P less than 0.001). Secretory phase type 2 activity was similar in all the women investigated. CONCLUSION: These data suggest that phospholipase A2 type 2 activity is increased in proliferative phase endometrium of women with polycystic ovaries but that the increase is not associated with chronic pelvic venous congestion.

17 beta-Hydroxysteroid oxidoreductase activity in the endometrium of normal women and patients with pelvic pain and polycystic ovaries.

17 beta-Hydroxysteroid oxidoreductase (17-OHSD) activity in the endometrium of women with pelvic pain syndrome (PPS) and/or polycystic ovaries (PCO) was compared with that of a control group. In both groups there was a 10-fold increase in 17-OHSD activity in secretory phase tissue compared with that of the proliferative phase, measured by both oxidative and reduction pathways, and a highly significant correlation between the two directions (P less than 0.001). In normal subjects, the ratio of activity measured under oxidative conditions: reducing conditions, at all stages of the cycle except late proliferative phase, was 2.1-2.9. In the late proliferative phase the ratio was 5.5 which was significantly different from other stages of the cycle. Similar ratios were found for the PPS/PCO group (proliferative phase 2.5, secretory phase 5.6); these were also significantly different (P less than 0.01). On the basis of this study oestrogen metabolism in the endometrium of women with PPS and/or PCO appears to be no different from that of normal subjects. Measurement of enzyme activity in high speed soluble and particulate fractions of endometrial homogenate indicated the presence of two activities with different cofactor requirements. Gel filtration chromatography of the soluble fraction revealed a single peak of activity coincident with a molecular weight of 30 kDa with a strong preference for NAD + as cofactor. These preliminary findings suggest the presence of both soluble and particulate forms of 17-OHSD activity in the endometrium.

Urinary lactate dehydrogenase isoenzyme analysis in adult population.

This investigation was a systemic study on an adult population of urinary lactate dehydrogenase (LDH) isoenzyme analysis for the distinction between upper and lower urinary tract infections. The study included 160 urine samples from patients and healthy individuals. On the basis of clinical symptoms, urinary bacterial colony counts, renal function tests and radiologic findings, the adults were divided into pyelonephritis group, cystitis group, pelvic lesion group, and control group. This technique correctly identified 23 of 26 patients with pyelonephritis by the presence of elevated LDH-V (over 10 percent) and all of 12 patients with cystitis by the presence of elevated LDH-I (over 60 relative units) but low LDH-V (below 10 percent or lower than LDH-I). In the pelvic group, the results of eight patients were consistent with cystitis and four with pyelonephritis. Our study confirms the sensitivity and specificity of the LDH isoenzyme technique for the differential diagnosis of urinary tract infection on adult patients and is consistent with previous studies on pediatric patients. However, one should be cautious to interpret the results of LDH isoenzymogram before extra-urinary tract lesions are excluded.