Ultrasonographic scoring system for the diagnosis of adnexal torsion.

PURPOSE: To establish the utility of an ultrasonographic scoring system for the diagnosis of adnexal torsion. METHODS: We retrospectively analyzing 358 adnexal torsion cases. Using Pearson's χ2 test we determined whether ultrasonographic signs were significantly associated with adnexal torsion. Receiver operating characteristic curves were used to evaluate the diagnostic efficacy of the system. Ultimately by using binary logistic regression we established a precise and convenient scoring system. RESULTS: The torsion score was based on five criteria that were identified to be independently associated with adnexal torsion: (1) abnormal position of the index adnexa (odds ratio [OR], 2.311); (2) presence of a mass or cyst (OR, 3.495); (3) unilateral ovarian enlargement (OR, 3.051); (4) vascular pedicle twisting (OR, 2.105); and (5) peripheral hypervascularity of the corpus luteum with ovarian edema(encapsulating cyst sign) (OR, 4.164).patients with torsion who scored 0, have a predicted diagnosis rate of 20.9%; patients whose scores were 1,2 have a predicted probability of 41.8% and 66.15%, respectively. For patients with torsion scores of 3, 4, and 5, predicted diagnosis rates were 84.16%, 93.52%, and 98.27%, respectively. CONCLUSION: The ultrasonographic scoring system is feasible and precisely diagnoses adnexal torsion using ultrasound.

Histone deacetylase SIR2 in Toxoplasma gondii modulates functions of murine macrophages in vitro and protects mice against acute toxoplasmosis in vivo.

Silent information regulator 2 (SIR2) in histone deacetylase (HDAC) is particularly conserved and widely expressed in all eukaryotic cells. HDAC is a crucial post-translational modification protein regulating gene expression. In the present study, a Toxoplasma gondii (T. gondii) silent information regulator 2 (TgSIR2) gene in HDAC was cloned and the modulation effects of recombinant TgSIR2 (rTgSIR2) on murine Ana-1 macrophages were characterized in vitro. The results indicated that rTgSIR2 had a good capacity to eliminate T. gondii by promoting proliferation, apoptosis, and phagocytosis, and modulating the secretion of nitric oxide (NO), pro-inflammatory cytokines, and anti-inflammatory cytokines. In in vivo experiments, animals were immunized with recombinant TgSIR2, followed by a lethal dose of T. gondii RH strain challenge 14 days after the second immunization. As compared to the blank and control group, the animals immunized with rTgSIR2 could generate specific humoral responses, as demonstrated by the significantly high titers of total IgG and subclasses IgG1 and IgG2a. Significant increases of IFN-γ, IL-4, and IL-10 were seen, while no significant changes were detected in IL-17. The percentage of CD4+ and CD8+ T lymphocytes in animals immunized with rTgSIR2 significantly increased. A significantly long survival time was also observed in animals vaccinated with rTgSIR2 14 days after the last immunization. All these results clearly indicate that rTgSIR2 played an essential role in modulating host macrophages and offered the potential to develop a therapeutic strategy against T. gondii.

Crosstalk of HDAC4, PP1, and GSDMD in controlling pyroptosis.

Gasdermin D (GSDMD) functions as a pivotal executor of pyroptosis, eliciting cytokine secretion following cleavage by inflammatory caspases. However, the role of posttranslational modifications (PTMs) in GSDMD-mediated pyroptosis remains largely unexplored. In this study, we demonstrate that GSDMD can undergo acetylation at the Lysine 248 residue, and this acetylation enhances pyroptosis. We identify histone deacetylase 4 (HDAC4) as the specific deacetylase responsible for mediating GSDMD deacetylation, leading to the inhibition of pyroptosis both in vitro and in vivo. Deacetylation of GSDMD impairs its ubiquitination, resulting in the inhibition of pyroptosis. Intriguingly, phosphorylation of HDAC4 emerges as a critical regulatory mechanism promoting its ability to deacetylate GSDMD and suppress GSDMD-mediated pyroptosis. Additionally, we implicate Protein phosphatase 1 (PP1) catalytic subunits (PP1α and PP1γ) in the dephosphorylation of HDAC4, thereby nullifying its deacetylase activity on GSDMD. This study reveals a complex regulatory network involving HDAC4, PP1, and GSDMD. These findings provide valuable insights into the interplay among acetylation, ubiquitination, and phosphorylation in the regulation of pyroptosis, offering potential targets for further investigation in the field of inflammatory cell death.

[Clinical value of three-dimensional transvaginal sonography in diagnosis of septate uterus and prediction of adverse pregnancy outcome].

OBJECTIVE: To investigate clinical value of three-dimensional transvaginal sonography (3D-TVS) in the diagnosis of septate uterus and the relationship between its parameters and adverse pregnancy. METHODS: From Mar. 2010 to Sept. 2011, 73 patients (aged 23-35 years) with septate uterus who were diagnosed by 3D-TVS in Fujian Province Maternal and Child Health Hospital were enrolled in this retrospective study. The septum width, septum angle, septum length and the length of remaining uterine cavity were measured among the patients with subseptate uterus, and then, the distortion rate was calculated. The hysteroscopic surgery was used as the gold standard, and the diagnostic titer of 3D-TVS in the diagnosis of septate uterus was determined.Receiver operating characteristic curve (ROC) were plotted to evaluate the diagnostic titer of uterine parameters measured by 3D-TVS in predicting the adverse pregnancy outcome among patients with subseptate uterus.Univariate logistic regression was used to analyze the effectiveness of uterine parameters on adverse pregnancy outcome. RESULTS: Using hysteroscopic surgery as the gold standard, the coincidence rate of diagnosis of septate uterus by 3D-TVS was 94% (69/73) . Among the patients with septate uterus, 25% (17/69) were complete septate uterus, 75% (52/69) were subseptate uterus. Among patients with subseptate uterus, the septum length[(2.2 ± 0.6) cm] and distortion rate in patients with adverse pregnancy (0.60 ± 0.10) were significantly higher than those without adverse pregnancy [(1.5 ± 0.6) cm,0.43 ± 0.13, both P < 0.05]. However, no significant difference in the width, angle and length of septum were observed between the two groups (P > 0.05). The area under ROC curve (AUC) of septum length and distortion rate in determining adverse pregnancy were 0.833 (95%CI: 0.721-0.944) and 0.800 (95%CI: 0.671-0.929), respectively. The optimal cutoff point of septum length was 1.94 cm, with the sensitivity was 74.3% and the specificity was 76.5%; the optimal cutoff point of distortion rate was 0.48, with the sensitivity was 77.1% and the specificity was 76.5%. The expectation morbidity ratio of adverse pregnancy was 2.717, 3.067 and 0.514 by every adding level of septum length, distortion rate, and length of remaining uterine cavity, respectively. CONCLUSIONS: 3D-TVS showed high accuracy in diagnosis of septate uterus. The septum length and distortion rate may predict the risk of adverse pregnancy, and the value of them can be used for screening adverse pregnancy in clinical practice.

A polyethylene glycol enhanced ligation-triggered self-priming isothermal amplification for the detection of SARS-CoV-2 D614G mutation.

We presented a polyethylene glycol (PEG) enhanced ligation-triggered self-priming isothermal amplification (PEG-LSPA) for the detection D614G mutation in S-glycoprotein of SARS-CoV-2. PEG was employed to improve the ligation efficiency of this assay by constructing a molecular crowding environment. Two hairpin probes (H1 and H2) were designed to contain 18 nt and 20 nt target binding site at their 3' end and 5' end, respectively. In presence of target sequence, it complemented with H1 and H2 to trigger ligation by ligase under molecular crowding condition to form ligated H1-H2 duplex. Then 3' terminus of the H2 would be extended by DNA polymerase under isothermal conditions to form a longer extended hairpin (EHP1). 5' terminus of EHP1 with phosphorothioate (PS) modification could form hairpin structure due to the lower Tm value. The resulting 3' end overhang would also fold back as a new primer to initiate the next round of polymerization, resulting in the formation of a longer extended hairpin (EHP2) containing two target sequence domains. In the circle of LSPA, long extended hairpin (EHPx) containing numerous target sequence domains was produced. The resulting DNA products can be monitored in real-time fluorescence signaling. Our proposed assay owns an excellent linear range from 10 fM to 10 nM with a detection limit down to 4 fM. Thus, this work provides a potential isothermal amplification method for monitoring mutations in SARS-CoV-2 variants.

The effectiveness of uterine parametrics measured by three-dimensional transvaginal sonography in the diagnosis of uterine cavity distortions.

BACKGROUND: To analyze the application values of quantitative parametrics measured by three-dimensional transvaginal sonography (3D-TVS) in the diagnosis of uterine cavity distortions. METHODS: A retrospective analysis was performed on consecutive 113 patients with septate uteruses, as diagnosed consistently by both 3D-TVS and hysteroscopic surgery, between Jan 2016 and Jan 2018. The intrauterine quantitative parametrics were compared between an infertility group and a miscarriage group. Receiver operating characteristic (ROC) curve analysis and evaluation indicators of diagnostic tests were plotted to analyze the relationships between parametrics and uterine cavity distortions. RESULTS: There were no significant differences in septum width, septum length, uterine angle, remaining uterine cavity length, and distortion rate between the infertility and miscarriage groups (P>0.05). The distortion rate was positively correlated with septum length, but showed a negative correlation with uterine angle and remaining uterine cavity length (all P<0.05). The area under the curve (AUC) of septum length for predicting a distortion rate higher than 50% was 0.969; with a cutoff value of 2.15 cm, and a diagnostic accuracy of 97.14%, sensitivity was 81.80% and specificity was 98.90%, respectively. Patients with lower septum length coupled with a higher remaining uterine cavity length had a lower risk of uterine cavity distortion. CONCLUSIONS: Septum length was shown to have the most important effect on uterine cavity distortion of all the uterine parametrics measured by 3D-TVS. Patients with septum lengths higher than 2.15 cm indicated a distortion rate of more than 50%, and tend to experience adverse pregnancy outcomes.

IL­37 expression is decreased in patients with hyperhomocysteinemia and protects cells from inflammatory injury by homocysteine.

As a novel anti­inflammatory cytokine of the interleukin (IL)­1 family, IL­37 protects the human body from diseases characterized by excessive inflammation. The pathologic process of hyperhomocysteinemia (hHcy) is accompanied by persistent inflammation. However, little is known regarding the role of IL­37 in hHcy. In the present study, the levels of cytokines including IL­37, IL­1ß, IL­6 and tumor necrosis factor­α in the supernatant were detected by ELISA. mRNA and protein expression were detected by Reverse transcription­quantitative PCR and western blotting, respectively. LDH level was determined by ELISA and the cell viability was detected through CCK­8 kit. In the present study, mean serum IL­37 levels of patients with hHcy were 32.3% lower than those of controls (P<0.01). In peripheral blood mononuclear cells (PBMCs) from patients with hHcy, mean IL­37 mRNA expression was 73.5% lower (P<0.01) and IL­37 protein expression was 77.7% lower compared with that of healthy controls (P<0.01). Furthermore, the results demonstrated that exogenous homocysteine (Hcy) stimulation markedly downregulated the mRNA and protein expression levels of IL­37 in PBMCs in vitro. In 293T cells, overexpression of IL­37 restored the cell viability impaired by Hcy, and reduced the release of lactate dehydrogenase and the proinflammatory cytokines IL­1ß, IL­6 and tumor necrosis factor­α. In conclusion, IL­37 was downregulated by Hcy in vivo and in vitro, and IL­37 exhibited a protective role against cell injury induced by Hcy.

The 100 most-cited articles on prenatal diagnosis: A bibliometric analysis.

BACKGROUND: The number of citations a published article receives can be used to demonstrate its impact on a field of study. The objective of this study was to identify and characterize the 100 most-cited research articles (T100) published on prenatal diagnosis. METHODS: The Web of Science (WOS) database was searched for papers on prenatal diagnosis published between 1900 and 2018. The 100 most-cited original articles and reviews were recorded. Each eligible paper was reviewed for authors, journal name, year of publication, country, institution, total citations, citation density, H-index, research field, article type, and keywords. RESULTS: The T100 were published between 1972 and 2015 with a mean of 332.7 citations per paper (range: 196-1254). Most of the T100 were published between 1990 and 2005, in 35 journals led by New England Journal of Medicine (n = 14) followed by Lancet (n = 10), and Proceedings of The National Academy of Sciences of the United States of America (n = 8). Studies on method application, which promotes field development, were the majority article type. The team of Lo YM featured prominently in the field, and the United States of America, United Kingdom, and Hong Kong, China were the leading countries/regions. Frequency of cooperation was also highest among these 3 regions. Hierarchical cluster analysis produced 4 groups of keywords. CONCLUSION: Our analysis provides a historical perspective on scientific progress in prenatal diagnosis and may assist clinicians and researchers in assessing the quality of research over the past 50 years. It also provides concise information to guide future research.

Tripterygium wilfordii Glycosides Upregulate the New Anti-Inflammatory Cytokine IL-37 through ERK1/2 and p38 MAPK Signal Pathways.

As a Chinese traditional patent medicine, Tripterygium wilfordii glycosides (TWG) have been approved by the China State Food and Drug Administration (Z32021007) for autoimmune and inflammatory diseases. Application of TWG leads to significant decrease of the inflammatory cytokines, such as IL-6, IL-1ß, and TNF-α. However, little is known whether TWG could regulate the anti-inflammatory cytokines and what the mechanism is. Here, we found that TWG could induce the upregulation of IL-37 which is a new anti-inflammatory cytokine. Furthermore, the inhibitors of ERK1/2 and/or p38 MAPK pathways suppressed IL-37 expression induced by TWG, indicating that the two pathways took part in this process. In conclusion, TWG could upregulate the anti-inflammatory cytokine IL-37 and ERK1/2 and p38 MAPK signal pathways were involved in the upregulation of IL-37 induced by TWG. The results showed that TWG had a potent activity on promoting the expression of IL-37, a new anti-inflammatory cytokine, which help further understanding the anti-inflammatory mechanism for the clinical application of TWG in therapy of diseases.

[The expression of human IL-37 in E.coli and preparation and characterization of mouse anti-IL-37 antibody].

OBJECTIVE: To construct a prokaryotic expression system for interleukin-37 (IL-37) and prepare its polyclonal antibody. METHODS: The gene encoding mature interleukin-37 (IL-37m) was amplified by PCR and subcloned into prokaryotic expression vector pET28a. Then the recombinant plasmid pET28a/IL-37m was transformed into E.coli Rosetta and expressed under IPTG induction. The recombinant IL-37m was purified through Ni²âº;-NT agarose gel column and the purified recombinant IL-37m was used as immunogen to immunize the BALB/c mouse. The titer and specificity of the mouse anti-IL-37 antibody were analyzed by ELISA, Western blotting and immunohistochemical staining, respectively. RESULTS: The recombinant IL-37 was successfully expressed and purified, and the mouse anit-IL-37 antibody was successfully prepared. ELISA showed that the titer of the antiserum was 1:128 000. Western blot analysis revealed that the antibody reacted with IL-37 specifically. Immunohistochemical staining detection manifested the antibody could recognize the native IL-37. CONCLUSION: The mouse anti-IL-37 antibody with high titer and specificity was successfully prepared.

[Effect of the ethanol extracts of starfish Asterias amurensis on the levels of serum IL-4 and IFN-γ in mice].

AIM: To investigate the effect of the ethanol extracts of the starfish Asterias amurensis on the levels of serum IL-4 and IFN-γ in mice. METHODS: The whole bodies of the starfish were chopped and extracted with ethanol. The ethanol extracts were chromatographed on silica gel column. The separating fractions of the ethanol extracts were intraperitoneally injected into mice, respectively. The levels of serum IL-4 and IFN-γ in mice were detected by ELISA. RESULTS: The ethanol extracts from the starfish were separated through silica gel column chromatography to obtain 8 fractions (I-VIII). The high levels of IL-4 and IFN-γ were produced in serum of the mice injected with fractions III and VIII of the ethanol extracts from the starfish Asterias amurensis. CONCLUSION: The fractions III and VIIII separated from the ethanol extracts of the starfish Asterias amurensis can stimulate the mice to produce high lelves of IL-4 and IFN-γ, which has the characteristic of natural kill T (NKT) cells activator. It is suggests that there is the active substance that can activate NKT cells in the starfish Asterias amurensis.

[Preparation and characterization of rabbit antibody against human mast cell carboxypeptidase].

AIM: To prepare antibody against human mast cell carboxypeptidase (hMC-CP) by using recombinant hMC-CP expressed in E.coli, and to characterize the antibody. METHODS: hMC-CP was expressed in E.coli with L-Arabinose induction and purified through Ni-NTA column. The purified hMC-CP as immunogen was used to immunize rabbit. The titer and the specificity of the rabbit anti-hMC-CP antibody was analyzed by indirect ELISA and Western blot respectively. RESULTS: The hMC-CP was successfully expressed and purified. The polyconal anit-hMC-CP antibody was prepared by immunizing rabbit using the purified recombinant protein, The titer of the generated antiserum was 1:6 400 by ELISA. Western blot analysis showed that this antibody could bind with hMC-CP specifically. CONCLUSION: The rabbit anti-hMC-CP antibody with high titer and high specificity has been prepared by using purified recombinant hMC-CP as immunogen, which lays the foundation for further research on detection and function of hMC-CP.