HMGB1 regulates lipopolysaccharide-induced cellular dysfunction in HTR8/SVneo cells: Implications for the role of HMGB1 in unexplained spontaneous miscarriage.

INTRODUCTION: Approximately half of miscarriages are of an unknown aetiology and are likely characterized by aberrant inflammation at the uteroplacental interface. High mobility group box 1 (HMGB1) is a ubiquitous nuclear protein that participates in the pathological inflammatory response. The present study investigated the role of HMGB1 in inflammation-induced damage in trophoblasts and elucidated the underlying mechanism. METHODS: Immunohistochemistry, qRT-PCR and Western blotting were used to detect the expression of HMGB1 in early unexplained miscarriage and normal placentas. Lipopolysaccharide (LPS)-induced HTR8/SVneo cells were used as an in vitro model to mimic the aberrant inflammation at the uteroplacental interface of miscarriage. The expression of HMGB1 and the autophagy-related proteins LC3 and Beclin1 was detected using Western blotting. Autophagy was studied in villous tissues using immunofluorescence and Western blotting. Cell proliferation and migration were analysed. RESULTS: The expression level of HMGB1 in villous tissues with early unexplained miscarriage was significantly higher than the normal pregnancy group. The inhibition of HMGB1 in LPS-treated HTR8/SVneo cells decreased the expression of Beclin 1 and LC3, which promoted cell proliferation and migration. We found a high level of autophagy in miscarriage placentas. HMGB1 and autophagy inhibition reversed the proliferation and migration of LPS-induced HTR-8/SVneo cells. DISCUSSION: Our results demonstrated that HMGB1 participated in LPS-induced inflammation via autophagy and regulated trophoblast functions, such as cell proliferation and migration, to potentially participate in the pathogenesis of miscarriage.

In vitro model of production of antibodies; a new approach to reveal the presence of key bacteria in polymicrobial environments.

BACKGROUND: There is a rapid emergence of multiple resistant gram-negative bacteria due to overuse of antibiotics in the treatment of infections. Biofilms consist of polymicrobial communities that survive the host's defense system. The key bacteria in biofilms are slow growing and support an attachment and rapid growth of other microorganisms. Current antimicrobial strategies often fail due to poor diagnosis of key pathogens in biofilms. The study aims to develop anti-bacterial human antibodies in vitro from patients who had recently undergone a systemic infection by pathogenic bacteria and to use these antibodies as a tool for detecting bacteria in biofilms. METHODS: Lymphocytes were separated from whole blood of patients (n = 10) and stimulated with heat-killed bacteria to produce antibodies in vitro. The specificity of antibodies in recognizing the bacteria against which they were directed was evaluated by surface plasmon resonance system (SPR) and electron microscopy. The ulcer secretions from patients with chronic and acute leg ulcers and healthy controls were analyzed by the SPR system and the results were compared with culture studies. RESULTS: The produced antibodies recognized bacteria with high sensitivity (SPR). The antibodies against Enterococcus fecalis bound specifically to the microorganism in a bacterial co-culture that was visualized by electron microscopy. CONCLUSION: In the present work, a method for producing specific antibodies against bacteria is introduced to recognize bacterial components in body fluids of patients suffering from pathogenic biofilms. This diagnostic technique may be most useful in clinical microbiology and in the choice of antibiotics in the treatment of serious infections.

[Effects of oral contraceptive pretreatment on controlled ovarian hyperstimulation and outcomes of IVF-ET].

OBJECTIVE: To investigate the effects of oral contraceptive pretreatment (OCP) in patients with polycystic ovarian syndrome (PCOS) undergoing controlled ovarian hyperstimulation for IVF-ET. METHODS: We randomly divided 85 patients with P-COS undergoing IVF-ET into an OCP (n = 53) and a control group (n = 32), the former received OCP, while the latter did not before the cycle. We retrospectively analyzed the data of the patients for the ovulation promoting effect of OCP and its influence on the incidence of ovarian hyperstimulation syndrome (OHSS) and outcomes of IVF-ET. RESULTS: Compared with the control group, OCP significantly reduced the formation of ovarian cyst (P < 0.05), remarkably increased the duration of gonadotropin stimulation and consumption (P < 0.01) , and markedly raised the percentage of mature ova (87.92% vs 92.85%, P < 0.05). But no significant differences were observed between the two groups in the incidences of moderate and severe OHSS, number of retrieved oocytes, and rates of fertilization, miscarriage and clinical pregnancy. CONCLUSION: OCP for patients with PCOS can help to control the time of ovarian stimulation, improve the synchronism of follicular development, and increase the duration of gonadotropin stimulation and consumption, but cannot change the incidences of moderate and severe OHSS.