Factors affecting the accuracy and reliability of the measurement of anti-Müllerian hormone concentration in the clinic.

OBJECTIVE: We aimed to identify the factors that influence serum anti-Müllerian hormone (AMH) concentration measurements. METHODS: We collected serum samples between May and September 2018 and compared the effect on AMH concentration measured by ELISA of conditions including venepuncture, storage time, storage temperature, locations of the reaction microplate, and the use of the oral contraceptive pill and gonadotrophin-releasing hormone (GnRH). RESULTS: AMH concentration was not affected by food intake but was affected by haemolysis. It was also much higher in samples on the edge of the ELISA microtitre plate. AMH concentration increased after incubation at room temperature for 1 day, 4°C for 3 days, -20°C for 1 month and -40°C for 4 months, but no change occurred during storage at -80°C for 9 months. AMH concentration was high in patients following GnRH agonist treatment but was not affected by oral contraceptives. CONCLUSIONS: No fasting is required prior to AMH measurement. Placement of serum samples on the edge of microtitre plates affects the results of the AMH ELISA. If serum samples cannot be assayed immediately, it is best to store them at -80°C. Basal AMH concentration cannot be used as a measure of ovarian reserve after GnRH agonist treatment.

Assessment of Anti-Mullerian Hormone and Anti-Mullerian Hormone Type II Receptor Variants in Women with Repeated Implantation Failures.

Repeated implantation failure (RIF) is a common endocrine disease that causes female infertility and the etiology is unknown. The abnormal expression of key proteins and hormones at the maternal-fetal interface affected the maternal-fetal communication and leads to adverse pregnancy outcomes. The expression of anti-Mullerian hormone (AMH) and AMH receptor II (AMHRII) was observed in the endometrium. This study aimed to investigate the expression of AMH and AMHRII at the human endometrium, decidual tissue, and blastocyst. Furthermore, the expression of AMH and AMHRII were examined in the RIF patients using immunohistochemistry and quantitative real-time PCR to test the AMHRII expression. The results demonstrated that AMH and AMHRII were present in healthy endometrium and AMHRII was highly expressed in mid-luteal phase. In addition, AMHRII expression was detected throughout the pregnancy and AMHRII's highest expression was in the second trimester. AMHRII was expressed in the blastocysts; however, AMH was not observed. The positive expression rate for AMHRII was significantly higher in the endometrium from RIF. Estrogen receptor (ER), insulin-like growth factor binding protein 1(IGFBP1), and prolactin (PRL) were significantly less expressed in RIF with high expression of AMHRII. The apoptosis was significantly higher in patients with high expression of AMHRII than in patients with normal expression of AMHRII. Our data suggests that AMHRII had an effect on RIF via the AMH and AMHRII signaling pathway. It participated in the development of RIF by interfering with endometrial decidualization and apoptosis.

Rapid discovery of tyrosinase inhibitors from Sophora flavescens:significance for comprehensive utilization of its non-medicinal resources / 中国中药杂志

Sophorae Flavescentis Radix,derived from the root of Sophora flavescens in the Leguminosae family,has been widely used in the medicine,agriculture,animal husbandry,and daily chemical industry. A pharmacophore model-based method for rapid discovery of tyrosinase inhibitors( TIs) from S. flavescens was established by molecular docking under Lipinski rules,and verified by enzyme assays. Briefly,the chemical constituent database of S. flavescens( CDSF) was established based on the previous papers. Theoptimal pharmacophore model( OPM) was constructed by DS 2019 on the basis of known active TIs. Eighty-three hits predominated by flavonoids having higher fitting scores with OPM than the positive control were screened out,and subjected to molecular docking based on the three-dimensional structure of tyrosinase crystal protein. The potential TIs such as kurarinone and nor-kurarinone were rapidly discovered from the compounds with higher docking scores than the positive control under the Lipinski rules. The results were verified by the in vitro enzyme assays. The inhibition activities of tyrosinase from non-medicinal parts of S. flavescens were also tested to explore the relationship between the inhibition activity and chemical compositions. This study is expected to provide data support for the comprehensive application and development of S. flavescens and also a new method for the rapid discovery of active substances or functional constituents in the complex systems.

Targeting Lymphocyte Activation Gene 3 to Reverse T-Lymphocyte Dysfunction and Improve Survival in Murine Polymicrobial Sepsis.

BACKGROUND: Lymphocyte activation gene 3 (LAG-3) is one of the immune checkpoint molecules, negatively regulating the T-cell reactions. The present study investigated the role of LAG-3 in sepsis-induced T-lymphocyte disability. METHODS: Mice sepsis was induced by cecal ligation and puncture (CLP). LAG-3 expression on some immune cells were detected 24 hours after CLP. LAG-3 knockout and anti-LAG-3 antibody were applied to investigate the effects on the survival, bacterial clearance. Cytokine levels, T-cell counts, and the presence of apoptosis (in blood, spleen, and thymus) were also determined. In vitro T-cell apoptosis, interferon γ secretion, and proliferation were measured. The expression of interleukin 2 receptor on T cells was also determined after CLP. RESULTS: LAG-3 was up-regulated on CD4+/CD8+ T, CD19+ B, natural killer, CD4+CD25+ regulatory T cells and dendritic cells. Both LAG-3 knockout and anti-LAG-3 antibody had a positive effect on survival and on blood or peritoneal bacterial clearance in mice undergoing CLP. Cytokine levels and T-cell apoptosis decreased in anti-LAG-3 antibody-treated mice. Induced T-cell apoptosis decreased, whereas interferon γ secretion and proliferation were improved by anti-LAG-3 antibody in vitro. Interleukin 2 receptor was up-regulated on T cells in both wild-type and LAG-3-knockout mice undergoing CLP. CONCLUSIONS: LAG-3 knockout or anti-LAG-3 antibody blockade protected mice undergoing CLP from sepsis-associated immunodysfunction and may be a new target for the treatment.

Anti-Müllerian Hormone Regulates Stem Cell Factor via cAMP/PKA Signaling Pathway in Human Granulosa Cells by Inhibiting the Phosphorylation of CREB.

Anti-Müllerian hormone (AMH) downregulates the level of stem cell factor (SCF) via the cAMP/PKA signaling pathway in human granulosa cells (GCs). Little information is available on the molecular mechanism underlying the interaction. This study is aimed at determining whether AMH regulates expression of SCF via the cAMP-PKA-CREB signaling pathway in human GCs. In the present study, we verified the binding of cAMP-response element-binding protein (CREB) to promoter of SCF in human GCs. Furthermore, the effect of CREB was tested on the SCF promoter, and the site of CREB binding to SCF promoter was identified using truncations as well as assays of SCF-promoted mutation and CREB mutation. To investigate the correlation among AMH, SCF promoter, and CREB, pGL-Basic-SCF+CREB was transfected into overexpressed AMH GCs (AMH-high GCs), low expressed AMH GCs (AMH-low GCs), and normal GCs (GCs), respectively. Finally, immunofluorescence, double immunostaining, and Western blot were carried out in AMH-high and AMH-low GCs to confirm the AMH-mediated regulation of SCF expression by inhibiting the phosphorylation of CREB (pCREB) in GCs. Results indicated CREB interacted with SCF promoter and significantly enhanced the transcription level of SCF. The CREB binding site was localized at 318-321 bp of SCF gene promote. AMH inhibits the expression of SCF by phosphorylation of CREB via the PKA signaling pathway in GCs. These findings provide an in-depth understanding of the molecular mechanism underlying AMH suppressing the follicle growth, which would aid in the development of a novel therapy.

Analysis of atypical computed tomography features of primary small intestinal lymphoma / 中华消化杂志

Objective To investigate the atypical computed tomography (CT) features of primary small intestinal lymphoma (PSIL),and its correlation with pathology.Methods From July 2007 to June 2018,at Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,the clinical features and CT imaging data of 29 histopathologically diagnosed PSIL with atypical CT features were retrospectively analyzed.Results A total of 29 cases were all confirmed as Non-Hodgkin's lymphoma including 23 cases of B cell lymphoma and six cases of peripheral T cell lymphoma.In 24 PSIL patients,the intestinal wall was unevenly thickened.While five cases had intra-and extra-intestinal masses.Images of four PSIL patients showed heterogeneous density at unenhanced CT scan,five cases presented with heterogeneous mild to moderate enhancement and five cases demonstrated with obvious enhancement at portal venous phase.Multiple ulcers in mucosa were found in 20 cases,and obviously abnormal mucosal enhancement was found in five cases,and 13 cases showed rough serosa layer of intestinal wall and the fat gap around the intestinal wall disappeared.Adjacent organs were involved in four cases and intestinal obstruction occurred in eight cases.Conclusion The atypical imaging of PSIL can be heterogeneous density of the lesion,heterogeneous or obvious enhancement at enhanced scan,multiple ulcers on the mucosal surface,thickening of the mucosal surface,blurred peripheral fat space,involvement of adjacent organs and intestinal obstruction.

Analysis of atypical computed tomography features of primary small intestinal lymphoma / 中华消化杂志

Objective@#To investigate the atypical computed tomography (CT) features of primary small intestinal lymphoma (PSIL), and its correlation with pathology.@*Methods@#From July 2007 to June 2018, at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, the clinical features and CT imaging data of 29 histopathologically diagnosed PSIL with atypical CT features were retrospectively analyzed.@*Results@#A total of 29 cases were all confirmed as Non-Hodgkin′s lymphoma including 23 cases of B cell lymphoma and six cases of peripheral T cell lymphoma. In 24 PSIL patients, the intestinal wall was unevenly thickened. While five cases had intra- and extra-intestinal masses. Images of four PSIL patients showed heterogeneous density at unenhanced CT scan, five cases presented with heterogeneous mild to moderate enhancement and five cases demonstrated with obvious enhancement at portal venous phase. Multiple ulcers in mucosa were found in 20 cases, and obviously abnormal mucosal enhancement was found in five cases, and 13 cases showed rough serosa layer of intestinal wall and the fat gap around the intestinal wall disappeared. Adjacent organs were involved in four cases and intestinal obstruction occurred in eight cases.@*Conclusion@#The atypical imaging of PSIL can be heterogeneous density of the lesion, heterogeneous or obvious enhancement at enhanced scan, multiple ulcers on the mucosal surface, thickening of the mucosal surface, blurred peripheral fat space, involvement of adjacent organs and intestinal obstruction.

Imaging diagnosis of autoimmune pancreatitis / 临床肝胆病杂志

Autoimmune pancreatitis (AIP) is a special autoimmune-mediated pancreatitis with the clinical manifestation of obstructive jaundice, and it often has good response to steroids. At present, AIP is classified into type 1 and type 2. The imaging findings of AIP include diffuse or localized pancreatic parenchymal enlargement, and the former is also called the “sausage-shaped” appearance. Imaging examinations of AIP have their own advantages and disadvantages. Ultrasound, endoscopic retrograde cholangiopancreatography, endoscopic ultrasound, computed tomography, magnetic resonance imaging, PET-CT, and PET-MR help with diagnosis and differential diagnosis and can be used for the monitoring and follow-up of clinical outcomes.

[Seminal plasma anti-Müllerian hormone and inhibin B and serum inhibin B in predicting the outcome of routine IVF fertilization].

OBJECTIVE: To analyze the correlations of seminal plasma (sp) anti-Müllerian hormone (spAMH) and inhibin B (spINHB) and serum INHB (serINHB) with semen parameters in oligoasthenospermia patients and explore their value in predicting the outcome of routine in vitro fertilization (IVF). METHODS: We obtained the levels of spAMH, spINHB and serINHB as well as semen parameters from 88 infertile males undergoing IVF due to oligoasthenospermia or female uterine tubal factors from August 2016 to February 2017. Using the ROC curve and Pearson's correlation analysis, we examined the effects of the obtained parameters on the fertilization rate and assessed the correlation of the levels of spAMH, spINHB and serINHB with the semen parameters of the patients. RESULTS: Concerning the predictive value for the outcome of IVF, Pearson's correlation analysis showed that the area under the ROC curve (AUC) of spAMH was 0.807 (sensitivity = 84.6%, specificity = 76%, cut-off point = 3.529, P <0.001) and that of spINHB was 0.768 (sensitivity = 84.6%, specificity = 88.7%, cut-off point = 31.117, P = 0.002). The serINHB level was found positively correlated with sperm concentration (r = 0.346, P = 0.001), total sperm count (r = 0.378, P <0.001), sperm motility (r = 0.521, P <0.001), and the percentage of progressively motile sperm (r = 0.343, P = 0.001). CONCLUSIONS: The levels of spAMH and spINHB can be used as laboratory indexes to predict the fertilization rate of routine IVF and are correlated with semen parameters in oligoasthenospermia patients, while that of serINHB has a positive correlation with the semen parameters of the patients.

Cystatin C combined with carotid artery plaque score improves predictability of significant coronary ar-tery disease in patients with chest pain / 实用医学杂志

Objective To assess the value of plasma level of cystatin C(Cyst-C)and carotid artery plaque score(PS)in predicting significant coronary artery disease(SCAD)in patients with chest pain. Methods A total of 192 patients with chest pain were involved retrospectively. According to the coronary angiography results ,the patients were divided into groups of SCAD (n = 128) and non-significant coronary artery disease (NSCAD , n = 64). Analyses were done to discuss the association of Cyst-C and PS with SCAD and the predictive value of Cyst-C and PS for SCAD. Results Logistic regression analysis demonstrated that Cyst-C and PS were independent predictors of SCAD. The odds ratios(OR)associated with the Cyst-C(each 1 mg/L)and PS(each 1 mm)for prediction of SCAD were 1.329 and 1.197,respectively. The areas under the receiver-operating characteristic curves(AUC)for the Cyst-C and the PS to predict the SCAD were 0.654 and 0.688,respectively. The combination of Cyst-C and PS increased the AUC to 0.742. The optimal cut-off value of Cyst-C was 0.95 mg/L and had a sensitivity of 72.3% for SCAD. Similarly,the optimal cut-off level of PS was 3mm which presented a sensitivity of 70.7%. A Cyst-C ≥ 0.95 mg/L and a PS ≥ 3 mm had negative predictive values of 46.3% and 48.3%,respectively,for SCAD. By combining Cyst-C with PS ,the sensitivity and negative predictive value increased to 83.6%and 62.5%, respectively. Conclusions Cyst-C and PS are both correlated with SCAD. They are independent predictive factors for SCAD in patients with chest pain. Combination of Cyst-C and PS can improve the predictability ,which may increase the reliability of screening SCAD before cardiac catheterization.