The use of IVF/ICSI and risk of postpartum hemorrhage: A retrospective cohort study of 153,765 women in China.

Objective: Postpartum hemorrhage (PPH) is the leading cause of maternal morbidity and mortality. Identifying women who are at high risk of PPH is crucial for implementing early preventive and interventive strategies. This study aimed to examine whether there is an association between the use of in vitro fertilization (IVF) /intracytoplasmic sperm injection (ICSI) and increased risk of PPH. Method: This retrospective cohort study was conducted using medical record data from women who delivered at a tertiary hospital in Shanghai, China, between January 1, 2013 and April 30, 2019. Logistic regression analysis was used to estimate the associations between the use of IVF/ICSI and the risk of PPH. Results: A total of 153,765 pregnant women were included, of which 6,484 conceived through IVF/ICSI and147,281 conceived naturally. The incidence of PPH was 1.9% in this cohort. The incidence of PPH in women who conceived through IVF/ICSI was significantly higher than those in women who conceived naturally (3.4% vs. 1.7%, p < 0.01). The use of IVF/ICSI was associated with an increase in the amount of postpartum blood loss. Compared to women who conceived naturally, the average amount of postpartum blood loss increased by 42.1 mL (ß = 42.1, 95% CI, 38.2-46.0) for women who conceived through IVF/ICSI. In addition, women who conceived through IVF/ICSI were at higher risk of maternal PPH. The adjusted odds ratio (OR) of PPH in women who conceived through ART was 2.7 (OR = 2.7, 95% CI, 2.3-3.1). Conclusion: Our findings demonstrated that women who conceived through IVF/ICSI were at higher risk of PPH and suggested to obstetricians and midwives to identify and implement early preventative strategies for PPH among pregnant women who conceived through IVF/ICSI.

Vaccination with Neospora caninum-cyclophilin and -profilin confers partial protection against experimental neosporosis-induced abortion in sheep.

The purpose of this study was to evaluate the protective efficacy of a vaccine consisting of recombinant Neospora caninum-cyclophilin (NcCyP) and -profilin (NcPro) in sheep. At 42 d and 21 d prior to mating, adult Dorset ewes were immunized with the rNcCyP-rNcPro vaccine (Group 1) or co-purifying non-recombinant (NR) control vaccine (Group 2). At 90 days post-mating, all immunized ewes and were challenged by intravenous injection with 106Nesopora caninum Illinois tachyzoites (NcTZ). Significant protection (P < 0.05) was observed in Group 1 with 9 out of 13 ewes giving birth to live-born lambs (69.2%), whereas all Group 2 ewes aborted (6/6). Neospora caninum was detected by PCR in both fetal and placental tissues from all Group 2 aborting ewes and in the placental tissues of Group 1 aborting ewes. In contrast, tissues and placentas of Group 1 live-born lambs were Neospora DNA-negative. Immunoreactive Neospora antigens were demonstrated in placentas associated with abortions, but not in tissues of aborted fetuses or those of the live-born lambs and their associated placentas. Anti-NcCyP and anti-NcPro titers were high in sera from Group 1 ewes and were further boosted by challenge infection, resulting in long-lasting (≥14.5 mos.) elevated titers. Lambs born to Group 1 ewes also had high NcCyP and NcPro titers in pre-colostrum sera. Immunofluorescence staining (IFA) of NcTZ with Group 1 post-immunization sera revealed both surface and internal TZ staining, a pattern consistent with that observed with rabbit sera to rNcCyP or rNcPro. Infection of NR-vaccinated ewes produced high but transient anti-NcCyP and anti-NcPro Ab titers. The results indicate that the NcCyP-NcPro vaccine elicited strong anti-N. caninum responses and conferred significant protection against abortion and transplacental transmission of N. caninum TZ in sheep.

Inhibition of Lipopolysaccharide (LPS)-induced inflammatory responses by selenium in bovine mammary epithelial cells in primary culture.

Selenium, in the form of selenoproteins, plays a pivotal role in anti-inflammatory processes and antioxidant defense system. The aim of this study was to examine the effects of selenium on lipopolysaccharide (LPS)-induced inflammatory responses in bovine mammary epithelial cells (bMEC) and to investigate the potential mechanism. bMEC viability was measured by MTT assay. TNF-α, IL-1ß, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) expressions were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). The activation of nuclear factor-kappa B (NF-κB) was determined by Western blotting. The results showed that the mRNA expressions of these inflammatory factors were significantly inhibited by selenium in a dose-dependent manner. At protein levels, Western blot analysis demonstrated that selenium dose-dependently decreased NF-κB p65 translocating from the cytoplasm to the nucleus. Taken together, these results suggest that the anti-inflammatory property of selenium in LPS-stimulated primary bMEC may be attributed to the downregulation of NF-κB activation.

Complete Genome Sequence of CHYJ130330, a Highly Virulent Strain of Porcine Epidemic Diarrhea Virus in South China.

Porcine epidemic diarrhea virus (PEDV) strain CHYJ130330 was isolated from southern China and shown to be highly virulent when inoculated into neonatal pigs. This report describes the complete genome sequence of CHYJ130330. These data will provide important insights into the variation of PEDV in China.

Protective effect of carvacrol on acute lung injury induced by lipopolysaccharide in mice.

Carvacrol, the major component of Plectranthus amboinicus, has been known to exhibit anti-inflammatory activities. The aim of this study was to investigate the effects of carvacrol on lipopolysaccharide (LPS)-induced endotoxemia and acute lung injury (ALI) in mice. Mice were injected intraperitoneally (i.p.) with LPS and the mortality of mice for 7 days were observed twice a day. Meanwhile, the protective effect of carvacrol (20, 40 or 80 mg/kg) on LPS-induced endotoxemia were detected. Using an experimental model of LPS-induced ALI, we examined the effect of carvacrol in resolving lung injury. The results showed that carvacrol could improve survival during lethal endotoxemia and attenuate LPS-induced ALI in mice. The anti-inflammatory mechanisms of carvacrol may be due to its ability to inhibit NF-κB and MAPKs signaling pathways, thereby inhibiting inflammatory cytokines TNF-α, IL-6 and IL-1ß production.

Magnolol inhibits lipopolysaccharide-induced inflammatory response by interfering with TLR4 mediated NF-κB and MAPKs signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Magnolia officinalis as a traditional Chinese herb has long been used for the treatment of anxiety, cough, headache and allergic diseases, and also have been used in traditional Chinese medicine to treat a variety of mental disorders including depression. AIM OF THE STUDY: Magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, has been reported to have anti-inflammatory properties. However, the underlying molecular mechanisms are not well understood. The aim of this study was to investigate the molecular mechanism of magnolol in modifying lipopolysaccharide (LPS)-induced signal pathways in RAW264.7 cells. MATERIAL AND METHODS: The purity of magnolol was determined by high performance liquid chromatography. RAW264.7 cells were stimulated with LPS in the presence or absence of magnolol. The expression of proinflammatory cytokines were determined by ELISA and reverse transcription-PCR. Nuclear factor-κB (NF-κB), inhibitory kappa B (IκBα) protein, p38, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and Toll-like receptor 4 (TLR4) were determined by Western blot. Further analyses were performed on mTLR4 and mMD2 co-transfected HEK293 cells. RESULTS: The result showed that the purity of magnolol used in this study was 100%. Magnolol inhibited the expression of TNF-α, IL-6 and IL-1ß in LPS-stimulated RAW264.7 cells in a dose-dependent manner. Western blot analysis showed that magnolol suppressed LPS-induced NF-κB activation, IκBα degradation, phosphorylation of ERK, JNK and P38. Magnolol could significantly down-regulated the expression of TLR4 stimulating by LPS. Furthermore, magnolol suppressed LPS-induced IL-8 production in HEK293-mTLR4/MD-2 cells. CONCLUSIONS: Our results suggest that magnolol exerts an anti-inflammatory property by down-regulated the expression of TLR4 up-regulated by LPS, thereby attenuating TLR4 mediated the activation of NF-κB and MAPK signaling and the release of pro-inflammatory cytokines. These findings suggest that magnolol may be a therapeutic agent against inflammatory diseases.

Lipopolysaccharide increases Toll-like receptor 4 and downstream Toll-like receptor signaling molecules expression in bovine endometrial epithelial cells.

The endometrium is easily contaminated with bacteria and the endometrial epithelial cells (EECs) play an important role in defence against invading pathogens which recognized pathogen-associated molecular patterns (PAMPs) via pattern recognition receptors (PRRs). Toll-like receptor 4 (TLR4) can recognize lipopolysaccharide (LPS) from Gram-negative bacteria and initiates innate immune responses. In this study, we stimulated bovine EECs with LPS from Escherichia coli (E. coli). The expression of TLR4 was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The expression of downstream TLR4 signaling molecules was detected by qRT-PCR. The results showed that TLR4 and downstream adaptor molecules, transcription factors and cytokines were up-regulated when bovine EECs were stimulated with LPS. Furthermore, the expression of TOLLIP and ß-defensin 5 were up-regulated when cells were stimulated with LPS. The results demonstrated that both MyD88 dependent and independent pathways in TLR4 were activated by LPS in bovine EECs. Bovine EECs have the immune repertoires required in defending against E. coli and play an important role in innate immune defence of the bovine endometrium.

Characterization of Neospora caninum microneme protein 10 (NcMIC10) and its potential use as a diagnostic marker for neosporosis.

Improvements in the serological diagnosis of neosporosis are needed to differentiate acute versus chronic Neospora caninum infections. In the present study, N. caninum microneme protein 10 (NcMIC10), similar to other microneme proteins, was shown to be released in a calcium-dependent manner. NcMIC10 may be discharged during active invasion of host cells by the parasite, and thus represent an excellent marker for the diagnosis of neosporosis. In order to test this hypothesis, recombinant NcMIC10 (rNcMIC10) was expressed in Escherichia coli, and polyclonal antibodies were generated against non-overlapping fragments of the protein. A capture ELISA was developed using these antibodies, and was found to be highly accurate and reproducible with a detection range of 10-10,000 pg/ml. The anti-rNcMIC10 antibodies used in this study did not cross-react with the Toxoplasma gondii antigens. NcMIC10 was detected by the ELISA in sera of 9 out of 10 goats (90%) experimentally infected with N. caninum tachyzoites. In general, goats infected with a lower dose (10(4)) of the parasite displayed a peak in NcMIC10 levels between weeks 4 and 5 post infection. Goats infected with a higher parasite dose (10(6)) displayed a more rapid increase in NcMIC10 levels. In most animals, NcMIC10 decreased to undetectable levels by week 6 post infection. This is the first circulating Neospora antigen-based assay which may complement the existing antibody-based assays for a rapid and cost-effective definitive diagnosis of neosporosis in livestock.

Neospora caninum tachyzoite- and antigen-stimulated cytokine production by bone marrow-derived dendritic cells and spleen cells of naive BALB/c mice.

Neospora caninum is an intracellular protozoan pathogen that causes abortion in cattle. This parasite elicits a typical type 1 immune response in host animals, and it is widely believed that the strong type 1 immune response during pregnancy may result in fetal death. Pro-inflammatory and/or inflammatory cytokines produced during either primary or secondary pathogen exposure are supposed to be the mediators of abortion. The present study defined cytokine production by murine naïve dendritic cells and spleen cells in response to whole Neospora tachyzoites (live, heat-killed, freeze-killed) or whole-cell tachyzoite lysate in the form of total (NcAg), soluble (sNcAg), or insoluble antigen (isNcAg). All tachyzoite and antigen preparations at high doses stimulated high levels of interleukin (IL) -12, interferon (IFN) -gamma, and tumor necrosis factor (TNF) -alpha, except for heat-killed tachyzoites and sNcAg, which induced moderate level of IL-12 and very low levels of IFN-gamma and TNF-alpha. In general, whole N. caninum tachyzoites were more effective in inducing IL-12, IFN-gamma, and TNF-alpha than the lysate antigen preparations. It appears that the heat-killed N. caninum tachyzoites were less potent in eliciting IFN-gamma or IL-10, but more effective in inducing IL-4. Thus, heat-inactivated tachyzoites or sNcAg alone may not be powerful enough to elicit strong type 1 immune responses against the disease. The present study comprehensively studied the production of critical cytokine by the murine dendritic cells and spleen cells in response to N. caninum; these results may facilitate a better understanding of antigen priming and aid in the design of vaccines/adjuvants against neosporosis.

Neospora caninum: cloning and expression of a gene coding for cytokine-inducing profilin.

Profilins are actin-binding proteins that in Toxoplasma gondii stimulate innate immunity in mice by binding Toll-like receptors (TLR) on dendritic cells (DC) leading to release of inflammatory cytokines, primarily IL-12 and IFN-gamma. The purpose of the present study was to characterize Neospora caninum profilin, termed NcProfilin. Recombinant NcProfilin was purified by affinity chromatography, and used to prepare specific antisera to allow characterization of native NcProfilin antigen in N. caninum tachyzoites. By immunoblotting, recombinant NcProfilin is 22kDa, and is similar in size to the respective 22kDa native protein. Immunofluorescence and immunoelectron microscopy localized native NcProfilin to the apical end of N. caninum tachyzoites. Incubation of recombinant NcProfilin with spleen cells from BALB/c mice induced release of IFN-gamma. Also, injection of BALB/c mice with purified rNcProfilin elicited a strong IFN-gamma and IL-12 responses at 6 and 24h after injection indicating that NcProfilin may be an important protein in regulation of cytokine responses to N. caninum.