Efficacy and safety of oral ibrexafungerp in Chinese patients with vulvovaginal candidiasis: a phase III, randomized, double-blind study.

PURPOSE: To evaluate the efficacy and safety of oral ibrexafungerp (HS-10366) versus placebo in Chinese patients with vulvovaginal candidiasis (VVC). METHODS: A double-blind, placebo-controlled, randomized, multicenter phase III study was conducted in symptomatic VVC patients. Patients received (2:1) twice-daily oral ibrexafungerp 300 mg or matching placebo for 1 day. The primary endpoint was clinical cure (vulvovaginal signs and symptoms [VSS] score = 0) at test-of-cure (TOC) on day 11 ± 3. The secondary endpoints included mycological eradication, overall response, and clinical improvement (VSS score ≤ 1) at TOC, and vulvovaginal symptom resolution at follow-up on day 25 ± 4. RESULTS: In total, 360 patients were included in the modified intention-to-treat set (defined as positive Candida cultured and receiving at least one study drug; 239 for ibrexafungerp, 121 for placebo). Compared with placebo, patients receiving ibrexafungerp had a significantly higher proportion of clinical cure (51.0% vs. 25.6%), mycological eradication (55.6% vs. 18.2%), overall response (33.9%, vs. 8.3%) at TOC and complete symptom resolution (74.5% vs. 39.7%, all P < 0.001) at follow-up. Subgroup analysis of clinical cure indicated that patients with C. albicans could benefit from ibrexafungerp over placebo. A similar benefit trend was also observed in those with non-albicans Candida by post-hoc analysis. Further analyses revealed similar efficacy of ibrexafungerp between patients with fluconazole non-susceptible C. albicans and fluconazole susceptible C. albicans regarding clinical cure and mycological eradication. Ibrexafungerp was generally well tolerated. Adverse events were primarily gastrointestinal and were mainly mild in severity. CONCLUSIONS: As a first-in-class antifungal agent, ibrexafungerp demonstrated promising efficacy and favorable safety for VVC treatment in Chinese patients. CHINADRUGTRIALS.ORG. CN REGISTRY NUMBER: CTR20220918.

The cellular trajectories of tumor-associated macrophages decipher the heterogeneity of pancreatic cancer.

Emerging evidence indicates that the interactions and dynamic changes among tumor-associated macrophages (TAMs) are pivotal in molding the tumor microenvironment (TME), thereby influencing diverse clinical outcomes. However, the potential clinical ramifications of these evolutionary shifts in tumor-associated macrophages within pancreatic adenocarcinoma (PAAD) remain largely unexamined. Single-cell RNA sequencing (scRNA-seq) data were retrieved from the Tumor Immune Single-cell Hub. The Seurat and Monocle algorithms were employed to elucidate the progression of TAMs, using non-negative matrix factorization (NMF) to determine molecular classifications. Subsequently, the prognosis, biological characteristics, genomic modifications, and immune landscape across various clusters were interpreted. Furthermore, the sensitivity of potential therapeutic drugs between subtypes was predicted. Cellular experiments were conducted to explore the function of the NR1H3 gene in pancreatic cancer. These experiments encompassed gene knockdown, proliferation assessment, clone formation evaluation, transwell examination, and apoptosis analysis. Trajectory gene expression analysis of tumor-associated macrophages identified three disparate clusters, each associated with different clinical outcomes Compared to clusters C1 and C2, cluster C3 is seemingly at a less advanced pathological stage and associates with a relatively favorable prognosis. Further investigation revealed pronounced genetic instability in cluster C2, whereas cluster C3 demonstrated notable genetic stability. Cluster C1, characterized as "immune-hot," exhibits an abundance of immune cells and elevated immune checkpoint expression, suggesting its suitability for immunotherapy. Furthermore, several potential therapeutic agents have been pinpointed, potentially facilitating the clinical application of these insights. Cell assays indicated that NR1H3 knockdown markedly induced apoptosis and suppressed clonogenesis, migration, and proliferation of pancreatic cancer cells in the PTAU-8988 and PANC-1 cell lines. Overall, our study discerned three clusters with unique characteristics, defined by the evolution of TAMs. We propose customized therapeutic strategies for patients within these specific clusters to improve clinical outcomes and optimize clinical management.

Talin1 regulates glucose metabolism and endometrial receptivity via GLUT-4 in patients with polycystic ovary syndrome and insulin resistance.

Objectives: Talin1 is a cytoskeletal protein and is localized between cells and the extracellular matrix. This study aimed to investigate the mechanism by which Talin1 affects glucose metabolism and endometrial receptivity via glucose transporter proteins-4 (GLUT-4) in patients with polycystic ovary syndrome (PCOS) and insulin resistance (IR). Methods: We examined the expression of Talin1 and GLUT4 in the receptive endometrium of PCOS-IR and control patients. GLUT4 expression was examined after silencing and overexpression of Talin1 in Ishikawa cells. We validated the interaction between Talin1 and GLUT-4 proteins using a co-immunoprecipitation (Co-IP) assay. After successfully establishing the C57BL/6j mouse model of PCOS-IR, the expression of Talin1 and GLUT-4 were examined in PCOS-IR and control mice. The effect of Talin1 on embryo implantation and the number of live births in mice were examined. Results: Our study found low expression of Talin1 and GLUT-4 in the receptive endometrium of PCOS-IR patients compared to that in control patients (p < 0.01). The level of GLUT-4 expression decreased after silencing Talin1 in Ishikawa cells and increased after overexpression of Talin1. Co-IP results showed that Talin1 interacts with GLUT-4 protein. We successfully established a PCOS-IR C57BL/6j mouse model and found that Talin1 and GLUT-4 expression in the receptive endometrium of PCOS-IR mice were lower than that in control mice (p < 0.05). In vivo experiments confirmed that the knockdown of Talin1 affects embryo implantation (p < 0.05) and live birth rate in mice (p < 0.01). Conclusions: Talin1 and GLUT-4 expression were decreased in the endometrium of PCOS-IR patients, and Talin1 may affect glucose metabolism and endometrial receptivity through GLUT4.

A nomogram to predict the risk of scar pregnancy after caesarean section.

The aim of this study was to identify the high-risk factors for caesarean scar pregnancy (CSP) and establish a nomogram to predict the risk of caesarean scar pregnancy in pregnant women with a history of caesarean section. Among 1273 pregnant women with a history of caesarean section, 70% of the patients (892 patients, training sample) were randomly selected for analysis, and a prediction model was generated. The remaining patients (381 patients, validation sample) were validated for the model. Four high-risk factors for CSP were established, including: parity, number of previous abortions, uterus position, and early vaginal bleeding. The area under the curve of the nomogram for the training set was 0.867 and that for the validation set was 0.881, indicating good performance. Calibration curves for predicting CSP showed good calibrations. Decision curve analyses showed good application prospects for the model. Our results show that our nomogram for predicting CSP risks can be a practical tool to help in the early identification of CSP.Impact StatementWhat is already known on this subject? The high-risk factors for "caesarean scar pregnancy", An simple nomogram could be constructed to predict the risk of the disease through these high-risk factors.What do the results of this study add? This study can quickly predict whether the patient is a high-risk group for uterine scar pregnancy based on the patient's previous pregnancy, early vaginal bleeding and uterine position.What are the implications of these findings for clinical practice and/or further research? Caesarean scar pregnancy was secondary Long-term complications after caesarean section that with a high risk of pregnancy. In this study, we established a nomogram based on the number of cases of CSP and a control group with a history of caesarean section delivery at term, The high-risk factors were assigned a certain risk value in the early stage, if the woman contains more high-risk factors, the higher the risk of developing CSP, it should be highly valued in the early stage, and the rate of visiting a doctor should be increased.

Three Major Gastrointestinal Cancers Could Be Distinguished through Subclass-Specific IgG Glycosylation.

Esophageal cancer (EC), gastric cancer (GC), and colorectal cancer (CRC) are three major digestive tract tumors with higher morbidity and mortality due to significant molecular heterogeneity. Altered IgG glycosylation has been observed in inflammatory activities and disease progression, and the IgG glycome profile could be used for disease stratification. However, IgG N-glycome profiles in these three cancers have not been systematically investigated. Herein, subclass-specific IgG glycosylation in CRC, GC, and EC was comprehensively characterized by liquid chromatography-tandem mass spectrometry. It was found that IgG1 sialylation was decreased in all three cancers, and the alterations in CRC and EC may be subclass-specific. IgG4 mono-galactosylation was increased in all three cancers, which was a subclass-specific change in all of them. Additionally, glycopeptides of IgG1-H5N5, IgG2-H4N3F1, and IgG4-H4N4F1 could distinguish all three cancer groups from controls with fair diagnostic performance. Furthermore, bioinformatics verified the differential expression of relevant glycosyltransferase genes in cancer progression. Significantly, those three gastrointestinal cancers could be distinguished from each other using subclass-specific IgG glycans. These findings demonstrated the spatial and temporal diversity of IgG N-glycome among digestive cancers, increasing our understanding of the molecular mechanisms of EC, GC, and CRC pathogenesis.

Distribution of tubal endometriosis: A 10-year retrospective study.

AIM: To investigate the distribution of tubal endometriosis (EM) in the right and left sides and four parts of the fallopian tube. METHODS: A retrospective, cross-sectional study was conducted on patients with tubal EM at the Fourth Affiliated Hospital of Guangxi Medical University from October 2011 to September 2021. Chi-square and binomial tests were used for analysis. RESULTS: Thirty-four patients (53.97%) had tubal resection due to EM (EM group). Twenty-nine patients (46.03%) had tubal resection due to non-EM (non-EM group). Thirty-two patients (50.80%) had left fallopian tube EM, 21 (33.33%) had right fallopian tube EM, and 10 (15.87%) had bilateral fallopian tube EM, with significant differences among them (p = 0.000). In the EM group, 15 patients (44.12%) had left fallopian tube EM, 13 (38.23%) had right fallopian tube EM, and 6 (17.65%) had bilateral fallopian tube EM (p = 0.052). In the non-EM group, statistically different (p = 0.001) diagnoses of left fallopian tube EM, right fallopian tube EM, and bilateral fallopian tube EM were 17 (58.62%), 8 (27.59%), and 4 (13.79%), respectively. In the EM group, 18 patients (52.94%) were in the ampullary region; 16 (47.06%) were in the nonampullary region (p = 0.864). In the non-EM group, 22 cases (75.86%) were in the ampullary region and 7 (24.14%) were in the nonampullary region, with a significant difference between them (p = 0.008). CONCLUSIONS: The incidence of left fallopian tube EM was higher than that of right and bilateral fallopian tube EM. The incidence of tubal ampullary EM was higher than that of nonampullary region.

The efficacy of intraoperative ICG fluorescence angiography on anastomotic leak after resection for colorectal cancer: a meta-analysis.

OBJECTIVE: The purpose of this study was to investigate whether intraoperative indocyanine green fluorescence angiography can reduce the incidence of anastomotic leak. METHODS: Present authors conducted a systematic search of PubMed, EMBASE, and Cochrane databases for randomized controlled trials (RCTs), prospective nonrandomized trials, and retrospective trials up to March 2020. Eleven papers fulfilling the screening criteria were included. INTERVENTION: Indocyanine green was injected intravenously after the division of the mesentery and colon but before anastomosis. The primary outcome measure was AL rate with at least 3 months of follow-up. Secondary outcome measure was operation time, postoperative complications, surgical site infection, reoperation, and ileus rate. The results were analyzed using STATA 12.0 software (Stata Corp, College Station, TX, USA). RESULT: A total of 3137 patients were collected in 11 studies. Meta-analysis showed that compared with conventional surgery, the ICG fluorescence angiography resulted in a fewer AL rate (OR = 0.31; 95% CI 0.21 to 0.44; P < 0.0001), postoperative complications (OR = 0.70; 95% CI 0.51 to 0.96; P < 0.025), and reoperation rate (OR = 0.334; 95% CI 0.16 to 0.68; P = 0.003). Operation time (weighted mean difference - 25.162 min; 95% CI - 58.7 to 8.375; P = 0.141), surgical site infection rate (OR = 1.11; 95% CI 0.59 to 2.09; P = 0.742) did not differ between the two groups. CONCLUSION: The result revealed that indocyanine green was associated with a lower anastomotic leakage rate after colorectal cancer resection. However, larger, multicentered, high-quality randomized controlled trials are needed to confirm the benefit of indocyanine green fluorescence angiography.

Meta-analysis of natural orifice specimen extraction versus conventional laparoscopy for colorectal cancer.

OBJECTIVE: To compare natural orifice specimen extraction surgery (NOSES) and conventional laparoscopic (LAP) surgery in treating colorectal cancer. METHODS: The present authors conducted a systematic search in the PubMed, EMBASE, and Cochrane databases for randomized controlled trials (RCTs), prospective nonrandomized studies, and retrospective studies up to May 2019. We used postoperative complications as the main endpoints, and used hospital stay, time to first flatus, operative time, postoperative pain, cosmetic result, wound infections, and oncological outcomes as the secondary endpoints. Subgroup analyses were conducted according to the different specimen extraction sites (transanal and transvaginal). A sensitivity analysis was carried out to evaluate the reliability of the outcomes. RevMan5.3 software was used for statistical analysis. RESULT: Twelve studies (one RCT, ten retrospective studies, and one prospective nonrandomized study) involving a total of 1437 patients (NOSES group 665 patients and LAP surgery group 772 patients) were included. Meta-analysis showed that compared with LAP surgery, NOSES resulted in a shorter hospital stay (WMD = -0.79 days; 95% CI -1.17 to -0.42; P < 0.001; P = 0.02), a shorter time to first flatus (WMD = -0.58 days; 95% CI -0.75 to -0.40; P < 0.001), less postoperative pain (WMD = -1.51; 95% CI -1.99 to -1.04; P < 0.001), a better cosmetic result (WMD = 1.37; 95% CI 0.59 to 2.14; P < 0.001), and fewer wound infections (OR = 0.13; 95% CI 0.05 to 0.35; P < 0.001) and postoperative complications (OR = 0.48; 95% CI 0.36 to 0.65; P < 0.001). Oncological outcomes did not differ between the two groups, while the operative time (WMD = 13.95 min; 95% CI 4.55 to 23.35; P = 0.004) was longer in the NOSES group. CONCLUSION: The present systematic meta-analysis is an attempt to assess the impact of NOSES, namely, its oncological outcomes and surgical safety in colorectal cancer patients. Pooled comparisons revealed that NOSES was superior to LAP surgery in terms of postoperative morbidity, postoperative pain, hospital stay, the time to first flatus, cosmetic results, and wound infections; however, NOSES was associated with a longer operative time. Considering the abovementioned limitations and the very low level of evidence of the comparisons, further RCTs are required to verify the results of our study.

Osthole inhibited cell proliferation and induced cell apoptosis through decreasing CPEB2 expression via up-regulating miR-424 in endometrial carcinoma.

Objective: Endometrial carcinoma (EC) was the fourth female malignancies in developed countries. Given that the prognosis of EC is extremely poor, it is vital to investigate its pathogenesis and effective therapeutic targets. However, the mechanism of osthole in EC remains unknown.Materials and methods: Firstly, the different doses of osthole (0, 50, 100, and 200 µM) were used to treat the Ishikawa and KLE cells. The cell proliferation, apoptosis, and cell cycle were measured by cell counting kit-8 (CCK-8), Annexin V-FITC/PI, and cell cycle assays. The apoptosis-related protein levels were examined by western blot. The miR-424 levels in Ishikawa and KLE cells were assessed by quantitative RT-PCR (qRT-PCR). Also, the binding of miR-424 and cytoplasmic polyadenylation element binding protein 2 (CEPB2) was detected by the luciferase reporter assay.Results: In this study, the increasing dose of osthole inhibited proliferation and induced apoptosis of Ishikawa and KLE cells. Moreover, the increasing dose of osthole up-regulated miR-424 and down-regulated the expression of CPEB2. CPEB2 was proved to be the target gene of miR-424. Interestingly, the over-expression of CPEB2 could reverse the changes of osthole-induced proliferation and apoptosis of Ishikawa and KLE cells.Conclusions: In summary, we provided first evidences that osthole inhibited proliferation and induced apoptosis through up-regulating miR-424 to inhibit expression of CPEB2 in EC. Our findings indicated that osthole might act as a novel and potential therapeutic agent for the treatment of EC.

Properties of High-Swelling Native Starch Treated by Heat-Moisture Treatment with Different Holding Times and Iterations.

Tapioca and potato starches were used to investigate the effect of heat-moisture treatment (HMT; 95-96 °C, 0-60 min, 1-6 iterations) on gelatinization properties, swelling power (SP), solubility and pasting properties. Tapioca starch had similar content and degree of polymerization of amylose, but a higher amylopectin short/long chain ratio, to potato starch. After HMT, the gelatinization temperature range was narrowed for tapioca starch, but was widened for potato starch. Decreases in SP and solubility were less for tapioca than potato starches, coinciding with a progressive shift to the moderate-swelling pasting profile for tapioca but a drastic change to the restricted-swelling profile for potato. Moreover, decreasing extents of SP and maximum viscosity for HMT tapioca starch were, respectively, in the range of 47-63% and 0-36%, and those of HMT potato starch were 89-92% and 63-94%. These findings indicate that the granule expansion and viscosity change of starch during gelatinization can be tailored stepwise by altering the HMT holding time and iteration.

Loss of Annexin A1 in macrophages restrains efferocytosis and remodels immune microenvironment in pancreatic cancer by activating the cGAS/STING pathway.

OBJECTIVE: Pancreatic cancer is an incurable malignant disease with extremely poor prognosis and a complex tumor microenvironment. We sought to characterize the role of Annexin A1 (ANXA1) in pancreatic cancer, including its ability to promote efferocytosis and antitumor immune responses. METHODS: The tumor expression of ANXA1 and cleaved Caspase-3 (c-Casp3) and numbers of tumor-infiltrating CD68+ macrophages in 151 cases of pancreatic cancer were examined by immunohistochemistry and immunofluorescence. The role of ANXA1 in pancreatic cancer was investigated using myeloid-specific ANXA1-knockout mice. The changes in tumor-infiltrating immune cell populations induced by ANXA1 deficiency in macrophages were assessed by single-cell RNA sequencing and flow cytometry. RESULTS: ANXA1 expression in pancreatic cancer patient samples correlated with the number of CD68+ macrophages. The percentage of ANXA1+ tumor-infiltrating macrophages negatively correlated with c-Casp3 expression and was significantly associated with worse survival. In mice, myeloid-specific ANXA1 deficiency inhibited tumor growth and was accompanied by the accumulation of apoptotic cells in pancreatic tumor tissue caused by inhibition of macrophage efferocytosis, which was dependent on cGAS-STING pathway-induced type I interferon signaling. ANXA1 deficiency significantly remodeled the intratumoral lymphocyte and macrophage compartments in tumor-bearing mice by increasing the number of effector T cells and pro-inflammatory macrophages. Furthermore, combination therapy of ANXA1 knockdown with gemcitabine and anti-programmed cell death protein-1 antibody resulted in synergistic inhibition of pancreatic tumor growth. CONCLUSION: This research uncovers a novel role of macrophage ANXA1 in pancreatic cancer. ANXA1-mediated regulation of efferocytosis by tumor-associated macrophages promotes antitumor immune response via STING signaling, suggesting potential treatment strategies for pancreatic cancer.

Observational study of surgical resection in small non-functional pancreatic neuroendocrine tumors: AS SEER-based study.

The potential benefits of surgical resection for small non-functional pancreatic neuroendocrine tumors (NF-PNETs) in terms of survival remain uncertain. This study aimed to evaluate the impact of surgical treatment on patients with NF-PNETs. Using SEER data, we identified 1102 patients from 2004 to 2015 with well and moderately differentiated pancreatic neuroendocrine tumors (PNETs). The associations between continuous variables and receipt of surgery were assessed using Wilcoxon rank-sum tests. Kaplan-Meier survival curves for OS were compared using the log-rank test. We compared outcomes in patients who received surgical resection with those in patients who did not, using a univariable Cox model with inverse probability weighting according to the propensity score and propensity-score matching. Among the cohort of 1102 patients, a majority of 965 individuals (87%) underwent surgical intervention. Upon conducting univariate analysis, we observed that surgical treatment significantly prolonged patients' survival [HR = 0.41, 95% CI [0.26-0.65] P < 0.001]. However, the old [HR = 3.27, 95% CI (2.24-4.76), P 0.001], male gender [HR = 1.82, 95% CI (1.23-2.68), P = 0.003], and moderately well-differentiated factors [HR = 1.71, 95% CI (1.04-2.80), P = 0.034] were found to potentially decrease patients' survival time. In the multivariate analysis, male gender [HR = 1.73, 95% CI (1.15-2.61), P = 0.009] and the old factor [HR = 3.52, 95% CI (2.33-5.31), P < 0.001] emerged as influential predictors with higher hazard ratios. Notably, surgical treatment remained a significant factor associated with improved overall survival [HR = 0.53, 95% CI (0.33-0.84), P = 0.007]. Propensity-score matching and inverse probability weighting were employed as analytical techniques. The univariate analysis results showed favorable outcomes in the weight group [HR = 0.48, 95% CI (0.29-0.78), P = 0.003] and matched group [HR = 0.44, 95% CI (0.22-0.85), P = 0.015], respectively. Survival analysis further confirmed that surgical treatment contributed to increased overall survival (log rank, P < 0.05) in both the matching and weight groups. Patients diagnosed with small, non-functioning pancreatic neuroendocrine tumors who undergo surgical intervention exhibit improved overall survival (OS) outcomes. Therefore, surgery is strongly recommended for this patient population.

Effects of letrozole combined with ethinylestradiol and cyproterone acetate tablets on serum sex hormones and lipid metabolism in patients with polycystic ovary syndrome.

OBJECTIVE: To investigate the effects of letrozole combined with ethinylestradiol and cyproterone acetate tablets on serum sex hormones and lipid metabolism in patients with polycystic ovary syndrome (PCOS). METHODS: Clinical data of 152 PCOS patients in the First Affiliated Hospital of Guangxi University of Chinese Medicine from May 2019 to June 2021 were collected for a retrospective analysis. Among the patients, 73 treated with ethinylestradiol and cyproterone acetate tablets alone were seen as control group (CG), and the rest 79 with letrozole combined with ethinylestradiol and cyproterone acetate tablets were seen as observation group (OG). The treatment efficacy was observed, and the adverse reactions in the course of treatment were counted. The levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone (T), estrogen (E2), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) were compared before and after treatment. The number of mature follicles, ovulation rate and pregnancy rate were assessed. Multivariate logistic regression analysis was used to detect the independent risk factors of ineffective efficacy. RESULTS: After the treatment, the total efficacy rate of the OG was higher than that of the CG (P<0.05); moreover, the levels of TC, TG, LDL, FSH, LH and T in OG were lower while HDL and E2 were higher (all P<0.05) than those of the CG. Also, the number of mature follicles, ovulation rate and pregnancy rate were higher in OG than those in the CG (all P<0.05). There was no obvious difference in the incidence of adverse reactions between the groups (P>0.05). Higher fasting glucose, higher Ferriman-Gallway hair score, single drug treatment regimen, higher systolic blood pressure, and lower E2 before treatment were independent risk factors for ineffective treatment efficacy. CONCLUSION: Letrozole combined with ethinylestradiol and cyproterone acetate tablets can enhance the treatment efficiency of PCOS and improve serum sex hormones and lipid metabolism in PCOS patients.

Comparison of the efficacy of oral contraceptives and levonorgestrel intrauterine system in intermenstrual bleeding caused by uterine niche.

This study aimed to compare the effectiveness of oral contraceptives and a levonorgestrel intrauterine system in treating intermenstrual bleeding due to uterine niche. We retrospectively analyzed 72 patients with intermenstrual bleeding due to uterine niche from January 2017 to December 2021, of whom 41 were treated with oral contraceptives and 31 with a levonorgestrel intrauterine system. Post-treatment follow-ups at 1, 3, and 6 months were conducted to compare the efficiency and adverse effects between the two groups. In the oral contraceptive group, the effectiveness rate was higher than 80% at 1- and 3-months post-treatment and higher than 90% at 6 months. In the levonorgestrel intrauterine system group, the effectiveness rates were 58.06%, 54.84%, and 61.29% at 1, 3, and 6 months of treatment, respectively. Oral contraceptives were more effective than the levonorgestrel intrauterine system in treating intermenstrual bleeding caused by uterine niche (p < 0.05).

High-dimensional single-cell analysis unveils distinct immune signatures of peripheral blood in patients with pancreatic ductal adenocarcinoma.

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies with poor response to immune checkpoint inhibitors. The mechanism of such poor response is not completely understood. Methods: We assessed T-cell receptor (TCR) repertoire and RNA expression at the single-cell level using high-dimensional sequencing of peripheral blood immune cells isolated from PDAC patients and from healthy human controls. We validated RNA-sequencing data by performing mass cytometry (CyTOF) and by measuring serum levels of multiple immune checkpoint proteins. Results: We found that proportions of T cells (CD45+CD3+) were decreased in PDAC patients compared to healthy controls, while proportion of myeloid cells was increased. The proportion of cytotoxic CD8+ T cells and the level of cytotoxicity per cell were increased in PDAC patients, with reduced TCR clonal diversity. We also found a significantly enriched S100A9+ monocyte population and an increased level of TIM-3 expression in immune cells of peripheral blood in PDAC patients. In addition, the serum level of soluble TIM-3 (sTIM-3) was significantly higher in PDAC patients compared to the non-PDAC participants and correlated with worse survival in two independent PDAC cohorts. Moreover, sTIM-3 exhibited a valuable role in diagnosis of PDAC, with sensitivity and specificity of about 80% in the training and validation groups, respectively. We further established an integrated model by combining sTIM-3 and carbohydrate antigen 19- 9 (CA19-9), which had an area under the curve of 0.974 and 0.992 in training and validation cohorts, respectively. Conclusion: Our RNA-seq and proteomic results provide valuable insight for understanding the immune cell composition of peripheral blood of patients with PDAC.

Single-cell RNA sequencing revealed subclonal heterogeneity and gene signatures of gemcitabine sensitivity in pancreatic cancer.

Introduction: Resistance to gemcitabine is common and critically limits its therapeutic efficacy in pancreatic ductal adenocarcinoma (PDAC). Methods: We constructed 17 patient-derived xenograft (PDX) models from PDAC patient samples and identified the most notable responder to gemcitabine by screening the PDX sets in vivo. To analyze tumor evolution and microenvironmental changes pre- and post-chemotherapy, single-cell RNA sequencing (scRNA-seq) was performed. Results: ScRNA-seq revealed that gemcitabine promoted the expansion of subclones associated with drug resistance and recruited macrophages related to tumor progression and metastasis. We further investigated the particular drug-resistant subclone and established a gemcitabine sensitivity gene panel (GSGP) (SLC46A1, PCSK1N, KRT7, CAV2, and LDHA), dividing PDAC patients into two groups to predict the overall survival (OS) in The Cancer Genome Atlas (TCGA) training dataset. The signature was successfully validated in three independent datasets. We also found that 5-GSGP predicted the sensitivity to gemcitabine in PDAC patients in the TCGA training dataset who were treated with gemcitabine. Discussion and conclusion: Our study provides new insight into the natural selection of tumor cell subclones and remodeling of tumor microenvironment (TME) cells induced by gemcitabine. We revealed a specific drug resistance subclone, and based on the characteristics of this subclone, we constructed a GSGP that can robustly predict gemcitabine sensitivity and prognosis in pancreatic cancer, which provides a theoretical basis for individualized clinical treatment.

Gαq-PKD/PKCµ signal regulating the nuclear export of HDAC5 to induce the IκB expression and limit the NF-κB-mediated inflammatory response essential for early pregnancy.

Decidualization, denoting the transformation of endometrial stromal cells into specialized decidual cells, is a prerequisite for normal embryo implantation and a successful pregnancy in human. Here, we demonstrated that knockout of Gαq lead to an aberrantly enhanced inflammatory state during decidualization. Furthermore, we showed that deficiency of Gαq resulted in over-activation of nuclear factor (NF)-κB signaling, due to the decreased expression of NFκBIA, which encode the IκB protein and is the negative regulator for NF-κB. Mechanistically, Gαq deficiency decreased the Protein kinase D (PKD, also called PKCµ) phosphorylation levels, leading to attenuated HDAC5 phosphorylation and thus its nuclear export. Aberrantly high level of nuclear HDAC5 retarded histone acetylation to inhibit the induced NFκBIA transcription during decidualization. Consistently, pharmacological activation of the PKD/PKCµ or inhibition of the HDAC5 restored the inflammatory state and proper decidual response. Finally, we disclosed that over-active inflammatory state in Gαq-deficient decidua deferred the blastocyst hatching and adhesion in vitro, and the decidual expression of Gαq was significantly lower in women with recurrent pregnancy loss compared with normal pregnancy. In brief, we showed here that Gαq as a key regulator of the inflammatory cytokine's expression and decidual homeostasis in response to differentiation cues, which is required for successful implantation and early pregnancy.

Excretion of Amyloid-ß in the Gastrointestinal Tract and Regulation by the Gut Microbiota.

BACKGROUND: Amyloid-ß (Aß) is important in the etiology of Alzheimer's disease (AD). Removal of Aß from the brain is a major strategy for the prevention and treatment of AD. OBJECTIVE: To clarify whether Aß42 can be cleared by intestinal excretion and whether the gut microbiota (GM) can affect the excretory clearance of Aß42 in the peripheral blood and intestines. METHODS: Male 8-month-old C57BL6 mice were maintained on either normal chow or received broad-spectrum antibiotics in their drinking water for one week. Sterile saline, fluorescein isothiocyanate (FITC), or FITC-Aß42 (fluorescein isothiocyanate-labeled amyloid-ß42 peptides) was injected 1 h before sampling. Related changes of Aß42 before and after injection were evaluated. RESULTS: FITC-Aß42 was injected into mice through the tail vein and could later be detected in feces. Furthermore, the fecal concentrations of FITC-Aß42 were higher in mice that had been fed antibiotics to alter their GM than in normal mice. However, the FITC-Aß42 concentrations in blood showed the opposite pattern. CONCLUSION: Aß42 can be excreted into the intestinal lumen and is regulated by the GM.

Exploration the global single-cell ecological landscape of adenomyosis-related cell clusters by single-cell RNA sequencing.

Background: Adenomyosis (AM) is a common benign uterine disease that threatens the normal life of patients. Cells associated with microenvironmental immune ecology are crucial in AM, although they are not as well understood at the cellular level. Methods: Single-cell sequencing (scRNA-seq) data were used to construct an AM global single-cell map, to further identify relevant cell clusters and infer chromosomal copy number variation (CNV) in AM samples. The biological functions of cell clusters were explored and cellular evolutionary processes were inferred by enrichment analysis and pseudotime analysis. In addition, a gene regulatory network (GRN) analysis was constructed to explore the regulatory role of transcription factors in AM progression. Results: We obtained the expression profiles of 42260 cells and identified 10 cell clusters. By comparing the differences in cell components between AM patients and controls, we found that significant abundance of endometrial cells (EC), epithelial cells (Ep), endothelial cells (En), and smooth muscle cells (SMC) in AM patients. Cell clusters with high CNV levels possessing tumour-like features existed in the ectopic endometrium samples. Moreover, the Ep clusters were significantly involved in leukocyte transendothelial cell migration and apoptosis, suggesting an association with cell apoptosis and migration. En clusters were mainly involved in pathways in cancer and apoptosis, indicating that En has certain malignant features. Conclusion: This study identified cell clusters with immune-related features, investigated the changes in the immune ecology of the microenvironment of these cells during AM, and provided a new strategy for the treatment of AM.

Epidermal growth factor induces a trophectoderm lineage transcriptome resembling that of human embryos during reconstruction of blastoids from extended pluripotent stem cells.

OBJECTIVES: This study aims to optimize the human extended pluripotent stem cell (EPSC) to trophectoderm (TE)-like cell induction with addition of EGF and improve the quality of the reconstructing blastoids. MATERIALS AND METHODS: TE-like cells were differentiated from human EPSCs. RNA-seq data analysis was performed to compare with TE-like cells from multiple human pluripotent stem cells (hPSCs) and embryos. A small-scale compound selection was performed for optimizing the TE-like cell induction and the efficiency was characterized using TE-lineage markers expression by immunofluorescence stanning. Blastoids were generated by using the optimized TE-like cells and the undifferentiated human EPSCs through three-dimensional culture system. Single-cell RNA sequencing was performed to investigate the lineage segregation of the optimized blastoids to human blastocysts. RESULTS: TE-like cells derived from human EPSCs exhibited similar transcriptome with TE cells from embryos. Additionally, TE-like cells from multiple naive hPSCs exhibited heterogeneous gene expression patterns and signalling pathways because of the incomplete silencing of naive-specific genes and loss of imprinting. Furthermore, with the addition of EGF, TE-like cells derived from human EPSCs enhanced the TE lineage-related signalling pathways and exhibited more similar transcriptome to human embryos. Through resembling with undifferentiated human EPSCs, we elevated the quality and efficiency of reconstructing blastoids and separated more lineage cells with precise temporal and spatial expression, especially the PE lineage. CONCLUSION: Addition of EGF enhanced TE lineage differentiation and human blastoids reconstruction. The optimized blastoids could be used as a blastocyst model for simulating early embryonic development.