Uterine natural killer (uNK) cells and recurrent miscarriage: a pilot randomised controlled trial of prednisolone in women with high uNK cells and recurrent miscarriage

Recurrent miscarriage (RM) is stressful. One reason for this is because no causes can be found for the pregnancy loss in the majority of cases. Focus has been on the endometrium which undergoes decidualisation in preparation for implantation. Any problems in the finely organised interactions between the endometrium and invading trophoblast cells may contribute towards a miscarriage. Immunological mechanisms are thought to be one of the pathways involved as there is the need of maternal adaptation of her immune response to the semi-allogenic developing embryo. Uterine natural killer (uNK) cells are the most abundant in the endometrium during the window of implantation. They interact with trophoblast cells, and are involved in vascular remodelling, an important step in implantation. Hence, they have a biological plausibility of playing a major role in RM. Both peripheral NK (pNK) and uNK cells tests have been developed as assessments of immunological causes of RM. A systematic review performed showed inadequate evidence for both pNK and uNK cells tests as markers for adverse pregnancy outcomes. There were only twelve studies, with 446 patients reporting pregnancy outcomes. There was no accepted consensus of normality and methodology for analysing NK cells. The conclusion was the need for well-designed studies to assess the role of NK cell tests as a clinically useful marker for screening. This led to the conduct of the pilot phase of a RCT of prednisolone in early pregnancy in women with idiopathic RM and raised uNK cells density. The main aim of this trial was to assess feasibility of recruitment and tolerability of prednisolone. Secondary clinical outcomes included live birth, types of miscarriage, miscarriage karyotype, gestational age at delivery, birthweight and pregnancy complications (eg: pre-eclampsia, gestational diabetes, fetal abnormality, stillbirth, IUGR). 160 women were screened for uNK cells density and 40 were randomised, despite the majority (85%) desiring prednisolone if given a choice. There was a trend towards improved live birth rate with prednisolone treatment but this was not significant. There were equal numbers of biochemical, sac and fetal pregnancy losses in both groups. All completed treatment with main reported side effects in the prednisolone group of insomnia. There were no pregnancy complications. The analysis of uNK cells was found to be very time consuming. To accommodate potentially large numbers who will be screened in the definitive trial, an alternative, quicker and equally accurate method of analysing uNK cells was developed using the colour deconvolution and area measurement plug-ins of a public domain image analysis package, Image J. Women supported this trial. Randomisation was acceptable. The prednisolone was safe. UNK cell density is a valid biomarker of severe outcomes. There was a trend towards improvement in live birth rates. This trial paves the way for the development of an endometrial based test to screen for the subgroup of women with RM that could potentially benefit from individualised treatment

Modulation of TRIB3 and Macrophage Phenotype to Attenuate Insulin Resistance After Downhill Running in Mice

Eccentric exercise training accompanied by a low-fat diet can prevent insulin resistance (IR) and is currently an effective method for the treatment of IR induced by high-fat diet (HFD)-associated obesity. However, the molecular mechanisms underlying this improvement of IR in adipose tissue are still not completely clear. In this study, 5–6-week-old male mice were randomly divided into a standard control diet (SCD) group (SC, n = 12) and a HFD group (HF, n = 72). After 12 weeks, 12 mice in each group were randomly sacrificed. The remaining mice in the HF group were randomly submitted to one of the following experimental protocols for 8 weeks: obesity-HFD-sedentary (OHF-Sed, n = 14), obesity-HFD-exercise (OHF-Ex, n = 16), obesity-SCD-sedentary (OSC-Sed, n = 14), and obesity-SCD-exercise (OSC-Ex, n = 16). All obese mice in the exercise group were subjected to downhill running. Half of the mice in each group received an insulin injection (0.75 U/kg) before sample collection. Epididymal fat was removed and weighed. Adipocyte size and inflammatory cell infiltration were observed by H&E staining. Both basal and insulin-stimulated GLUT4 fluorescence and protein contents were detected by immunofluorescence and Western blot. Levels of IL-1β and IL-10 were detected by ELISA. Protein contents of iNOS, Arg-1, TRIB3, p-AKT, and AKT were determined by Western blot. CD86 and CD206 fluorescence were determined by immunofluorescence. The results showed that a HFD for 12 weeks induced IR accompanied by adipose tissue macrophages M1 polarization (increased iNOS protein content and CD86 fluorescence) and TRIB3-AKT activation. Downhill running accompanied by a low-fat diet attenuated IR (p < 0.01), reduced inflammation levels (increased IL-10 protein content and decreased IL-1β protein content), inhibited adipose tissue macrophages M1 polarization (decreased iNOS protein content and CD86 fluorescence) and promoted M2 polarization (increased Arg-1 protein content and CD206 fluorescence), and suppressed TRIB3-AKT signaling. We concluded that downhill running accompanied by dietary fat regulation attenuates HFD-related IR in mice, which may be associated with reduced TRIB3-AKT signaling and activated M2 macrophages in adipose tissue

Comparative studies on the regulatory effects of raw and charred hawthorn on functional dyspepsia and intestinal flora

Purpose: To compare the effects of raw hawthorn (RH) and charred hawthorn (CH) on functional dyspepsia (FD) and intestinal flora (IF).Methods: A rat model of FD was established through use of a chronic stimulator. Rat models were evaluated by the rat’s physical state, body weight, diet, and histopathological examination. After RH or CH administration, the digestive function of the rats was evaluated by determining gastric emptying and intestinal propulsion rate, diversity of intestinal flora.Results: RH and CH both improved gastric emptying and intestinal propulsion rate in FD group when compared to control group (p < 0.05). CH yielded higher treatment effectiveness than RH. Sixteen phyla of microbiomes were recognized from all samples. After FD model establishment, the relative abundance of Lactobacillus, Lachnospiraceae and Bacteroidales decreased compared to normal control rats. On the other hand, the relative abundance of Helicobacter and Bacteroides in the model control group increased compared to normal control. After RH and CH treatment, the relative abundance of all dysregulated phyla was restored to varying degrees, but the levels after CH treatment were similar to those of the normal control group.Conclusion: The relative abundance of intestinal flora of FD model rats is significantly different from that of rats in normal control group. Thus, RH and CH intervention improves digestive function, and the mechanisms may be related to adjustment of gut dysbacteriosis.Keywords: Raw Hawthorn, Charred Hawthorn, Functional dyspepsia, Intestinal flor

Systematic Review and Meta-Analyses of The Interaction Between HIV Infection And COVID-19: Two Years’ Evidence Summary

Introduction During the COVID-19 pandemic, people living with HIV (PLWH) were considered to be at risk of worse COVID-19 outcomes once infected. However, the existing evidence is inconsistent. This systematic review and meta-analysis aimed to compare the risk of SARS-CoV-2 infection, severe COVID-19 symptoms, and mortality among PLWH and patients without HIV. Method The articles included studies published in PubMed, Medline, Embase, and Cochrane between December 1, 2019, and December 1, 2021. We included the original studies published in English focusing on observational studies assessing the risk of SARS-CoV-2 infection, severe COVID-19 symptoms, and mortality among PLWH. Four independent reviewers extracted data. STrengthening the Reporting of OBservational studies in Epidemiology-Modified (STROBE-M) checklist was used for quality assessment. For the results with heterogeneity I2 >75%, a random-effects model was employed. Otherwise, a fixed-effects model was used. The risk of SARS-CoV-2 infection, severe COVID-19 symptoms, and mortality were compared with and without HIV.ResultsWe included a total of 32 studies and 71,779,737 study samples, of whom 797,564 (1.11%) were PLWH. Compared with COVID-19 patients without HIV infection, PLWH had comparable risk of SARS-CoV-2 infection (adjusted Risk Ratio=1.07, 95% CI: 0.53-2.16, I2 = 96%, study n=6, n=20,199,805) and risk of developing severe COVID-19 symptoms (aRR=1.06, 95% CI: 0.97-1.16, I2 = 75%, n=10, n=2,243,370). PLWH, if infected with SARS-CoV-2, were found to have an increased risk of mortality compared with people without HIV (aRR=1.30, 95% CI: 1.09-1.56, I2 = 76%, study n=16, n=71,032,659). This finding was consistent across different subgroup analyses. Conclusion PLWH are at increased risk of COVID-19 related mortality once infected. The local health system should, on the one hand, strengthen COVID-19 prevention and clinical management among PLWH to avoid infection and, on the other hand, sustain the HIV care continuum for PLWH for HIV management

子宮頸部及び内膜における aromatase の局在と周期性変化

Objectives: To determine the role of maternal CYP1A1, GSTT1, and GSTM1 metabolic gene polymorphisms in modulating the association between pregnancy smoking exposure and fetal growth restriction. Study design: A case-control study was conducted to investigate if the association of pregnancy smoking and birth outcome was modulated by maternal gene polymorphisms. A total of 90 mothers with an IUGR baby (cases) and 180 mothers without IUGR (controls) were enrolled. Results: Almost half of smokers who carried a CYP1A1 variant (51.3%), GSTT1 null (43.6%), or GSTM1 null genotypes (64.1%) delivered a baby with IUGR. Smokers with the variant CYP1A1 "aa" genotype had babies with lower mean birthweight than non-smokers with the same genotype (p = 0.004). An interaction test showed increased prevalence of IUGR in smokers with the CYP1A1 (Aa/aa) variant (adjusted OR, 1.9; 95% CI, 1.4-5.5, p = 0.01), or with the GSTT1 null (AOR, 1.5; 1.1-3.1, p = 0.001), or GSTM1 null genotypes (AOR, 1.5; 1.2-3.7, p = 0.001). Conclusions: Risk of fetal growth restriction in mothers who smoked during pregnancy was modulated by maternal metabolic gene polymorphisms. The genetic control of the conversion of toxic metabolites of tobacco smoke to less damaging substances is important for maternal and fetal health. (C) 2008 Elsevier Ireland Ltd. All rights reserved

Comparative Proteomic Approach Identifies Pkm2 and Cofilin-1 as Potential Diagnostic, Prognostic and Therapeutic Targets for Pulmonary Adenocarcinoma

Lung cancer is the leading cause of cancer-related death in the world. Non-small cell lung carcinomas (Non-SCLC) account for almost 80% of lung cancers, of which 40% were adenocarcinomas. For a better understanding of the molecular mechanisms behind the development and progression of lung cancer, particularly lung adenocarcinoma, we have used proteomics technology to search for candidate prognostic and therapeutic targets in pulmonary adenocarcinoma. The protein profile changes between human pulmonary adenocarcinoma tissue and paired surrounding normal tissue were analyzed using two-dimensional polyacrylamide gel electrophoresis (2-DE) based approach. Differentially expressed protein-spots were identified with ESI-Q-TOF MS/MS instruments. As a result, thirty two differentially expressed proteins (over 2-fold, p<0.05) were identified in pulmonary adenocarcinoma compared to normal tissues. Among them, two proteins (PKM2 and cofilin-1), significantly up-regulated in adenocarcinoma, were selected for detailed analysis. Immunohistochemical examination indicated that enhanced expression of PKM2 and cofilin-1 were correlated with the severity of epithelial dysplasia, as well as a relatively poor prognosis. Knockdown of PKM2 expression by RNA interference led to a significant suppression of cell growth and induction of apoptosis in pulmonary adenocarcinoma SPC-A1 cells in vitro, and tumor growth inhibition in vivo xenograft model (P<0.05). In addition, the shRNA expressing plasmid targeting cofilin-1 significantly inhibited tumor metastases and prolonged survival in LL/2 metastatic model. While additional works are needed to elucidate the biological significance and molecular mechanisms of these altered proteins identified in this study, PKM2 and cofilin-1 may serve as potential diagnostic and prognostic biomarkers, as well as therapeutic targets for pulmonary adenocarcinoma

Role of miR-148a in Mitigating Hepatic Ischemia-Reperfusion Injury by Repressing the TLR4 Signaling Pathway via Targeting CaMKIIα in Vivo and in Vitro

Background/Aims: Hepatic ischemia-reperfusion (I/R) injury, which is mainly induced by inflammation and unstable intracellular ions, is a major negative consequence of surgery that compromises hepatic function. However, the exact mechanisms of liver I/R injury have not been determined. Positive crosstalk with the Ca2+/CaMKII pathway is required for complete activation of the TLR4 pathway and inflammation. We previously found that miR-148a, which decreased in abundance with increasing reperfusion time, targeted and repressed the expression of CaMKIIα. In the present study, we examined the role of the miR-148a machinery in I/R-induced Ca2+/CaMKII and TLR4 signaling changes, inflammation, and liver dysfunction in vivo and in vitro. Methods: Liver function was evaluated by serum aminotransferase levels and hematoxylin-eosin (HE) staining. Inflammatory factors were detected by enzyme-linked immunosorbent assay. Gene and protein expression were assessed by RT-PCR and western blot. Small interfering RNA was used to silence target gene expression. HE staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling were used to measure hepatic tissue apoptosis. These assays were performed to identify factors upregulated in hepatic I/R injury and downregulated by miR-148a. Results: We manifested that expression of CaMKIIα and phosphorylation of TAK1 and IRF3 were elevated in hypoxia/reoxygenation (H/R)-treated primary Kupffer cells (KCs) and liver tissue of I/R-treated mice, but these effects were attenuated by treatment with miR-148a mimic and were accompanied by the alleviation of liver dysfunction and hepatocellular apoptosis. Luciferase reporter experiments showed that miR148a suppressed luciferase activity by almost 60%. Moreover, knockdown of CaMKIIα in H/R KCs led to significant deficiencies in p-TAK1, P-IRF3, IL-6, and TNF-α, which was consistent with the effects of miR-148a overexpression. Otherwise, the same trend of activation of TAK1 and IRF3 and inflammatory factors in vitro was observed in the siTAK1 + siIRF3 group compared with the siCaMKIIα group. Conclusion: Taken together, we conclude that miR-148a may mitigate hepatic I/R injury by ameliorating TLR4-mediated inflammation via targeting CaMKIIα in vitro and in vivo

Fluorescent protein tagged hepatitis B virus capsid protein with long glycine-serine linker that supports nucleocapsid formation

Fusion core proteins of Hepatitis B virus can be used to study core protein functions or capsid trafficking. A problem in constructing fusion core proteins is functional impairment of the individual domains in these fusion proteins, might due to structural interference. We reported a method to construct fusion proteins of Hepatitis B virus core protein (HBc) in which the functions of fused domains were partially kept. This method follows two principles: (1) fuse heterogeneous proteins at the N terminus of HBc; (2) use long Glycine-serine linkers between the two domains. Using EGFP and RFP as examples, we showed that long flexible G4S linkers can effectively separate the two domains in function. Among these fusion proteins constructed, GFP-G4S186-HBc and RFP-G4S47-HBc showed the best efficiency in rescuing the replication of an HBV replicon deficient in the core protein expression, though both of the two fusion proteins failed to support the formation of the relaxed circular DNA. These fluorescent protein-tagged HBcs might help study related to HBc or capsids tracking in cells