Clinical Samples Research Articles

The effect of the COVID-19 pandemic and school closures on self-harming and suicidal behaviors in adolescents: A comprehensive meta-analysis.

The COVID-19 pandemic has significantly affected adolescents' mental health, raising concerns about the potential increase in self-harming and suicidal behaviors (SIB). This study aimed to evaluate the pandemic's impact on SIB among adolescents and explore the effects of school closures and psychiatric vulnerabilities to inform future interventions. A systematic review and meta-analysis were conducted using PubMed, resulting in 420 studies, of which 36 studies (comprising 53 samples) met the inclusion criteria, encompassing 1,754,106 adolescents. Effect sizes were calculated using proportion effect size. Heterogeneity across studies was assessed using Q-statistics and I², while Egger's test was applied to evaluate publication bias. Differences between clinical and community samples, as well as the influence of psychiatric symptoms or disorders, were examined. Clinical samples demonstrated higher rates of SIB compared to community samples, with adolescents having psychiatric symptoms or disorders being more vulnerable. Overall, the pandemic was associated with a decline in SIB rates compared to pre-pandemic levels. Suicidal ideation decreased by 4.14% in combined samples and by 7.13% in community samples. Self-harm rates fell by 5.38% and 6.62% in combined and community samples, respectively. In contrast, clinical samples showed a distinct trend, with suicidal ideation decreasing by 29.39%, while self-harm rates increased by 32.51% and suicide attempts rose by 22.41%. The COVID-19 pandemic's impact on adolescents' SIB varied significantly across population types and levels of psychiatric vulnerability. While general declines were observed, the pronounced increases in self-harm and suicide attempts among clinical samples highlight the need for tailored mental health interventions. Addressing these disparities is essential for future mental health strategies and crisis preparedness.

Pharmacological interventions for patients with chronic primary musculoskeletal pain: disparity between synthesized evidence and real-world clinical practice.

Chronic primary musculoskeletal pain (CPMP) poses a major problem of public health, with high prevalence rates and economic burden. There is a wealth of clinical trials examining pharmacological interventions for patients with CPMP. Nevertheless, evidence from such trials does not necessarily mirror clinical realities. We aimed to compare data sets from a clinical sample with an randomized controlled trial (RCT)-based sample. Both data sets included participants living with CPMP who received pharmacological interventions. The clinical sample was retrieved from electronic health records. The RCT-based sample stemmed from a network meta-analysis project. The following outcomes were used: demographic information, diagnosis-specific data, and pharmacological interventions (categorized according to the World Health Organization [WHO] analgesic ladder). The clinical sample consisted of 103 patients (mean age: 50.25 years; SD: 14.0) and the RCT-based samples contributed 8665 participants (mean age: 51.97 years; SD: 6.74). In both samples, the proportion of women was higher than that of men (ie, 74.8% vs 58.9%). Psychiatric disorders were the most common comorbidities in the clinic sample but also the most frequent reason for patient exclusion in RCTs. The 2 samples differed significantly in medication classified as WHO III (clinical sample: 12.9%; RCT sample: 23.5%; P = 0.023) and WHO IV (clinical sample: 23.4%; RCT sample: 8.6%; P < 0.001), yet not WHO I and II. Our findings suggest a disparity between research-based study populations and clinical populations with CPMP. We advocate for future investigations on how to implement robust scientific evidence into real-world clinical practice, with a particular focus on addressing psychiatric comorbidities.

Dimensionality, Sensitivity and Specificity of Different Versions of the Shirom-Melamed Burnout Questionnaire/Measure in Clinical and Non-Clinical Populations.

The Shirom-Melamed Burnout Questionnaire/Measure (SMBQ/SMBM) is a self-report instrument frequently used for assessing degree of burnout and screening for stress-related exhaustion disorder. The aim of the present study was three-fold. First, to examine reliability and construct validity of different versions of SMBM with 6-22 items in a clinical context. Second, to examine the criterion validity by assessing sensitivity and specificity and determining clinical cut-offs for these versions of the SMBM, and third to examine the prevalence of burnout in a general population and primary care sample using the proposed cut-offs. Two Swedish samples were used for the first two purposes: a clinical sample of patients diagnosed with exhaustion disorder (n=149), and a matched sample of healthy controls (n=60). For the third purpose a sample from the general population (n=3406), and a primary care clinical sample (n=326) was used. The modified versions of the SMBM showed good internal consistency, construct validity, dimensionality and model fit on the clinical exhaustion disorder sample, as well as configural measurement invariance across clinical and non-clinical samples. The sensitivity (94.6%-95.3%) and specificity (93.3%-95.0%) in identifying cases with exhaustion disorder based on the cut-off of 4.0 for the 19-, 16- and 11-items versions, and on the cut-off of 3.75 for the 6-item version was high. The prevalence of burnout was 81.2% in the primary care sample and 16.6% in the general population sample. The findings indicate that the SMBM is a useful instrument for screening for exhaustion disorder and burnout.

VG@nAu-based fluorescent biosensor for grading Alzheimer's disease by detecting P-tau181 protein in clinical samples.

Alzheimer's disease (AD) is a neurodegenerative disorder with a very long duration, posing a serious threat to people's life and health. To date, no medicine that can cure or reverse the disease has been developed or reported, so early diagnosis and timely intervention are essential. The concentration of Phosphorylated tau181 (P-tau181) in blood has been approved by FDA as a standard for assisting clinical diagnosis of AD. In this study, a fluorescence biosensor based on vertical graphene-modified nano-gold film (VG@nAu) was developed for the grading of AD diagnosis by detecting P-tau181 protein in the blood. The VG@nAu substrate that produces hundreds of times IR800 molecular compared with a glass substrate, which is prepared by sputtering nano-gold onto vertical graphene surface. The detection limit of this fluorescence biosensor for P-tau181 is 0.82pg/mL. The sensor also shows excellent selectivity and stability. Moreover, we analyzed the levels of P-tau181 in 16 clinical samples divided into four groups (HC group, SCD group, MCI group, and AD group) and found significant differences in P-tau181 levels. The grouping result based on the detection results using this biosensor is consistent with the grouping results of the clinical doctors. This study achieves ultrasensitive detection of P-tau181 protein by constructing a fluorescent biosensor, which validates the feasibility of the VG@nAu-based fluorescence biosensor for the grading of AD diagnosis. This research provides a promising tool for ultrasensitive detecting biomarkers in clinical samples, indicating significant potential for its use in related diagnosing disease.

Proliferation Inhibited by Genipin in Human Leukemia K562 Cells: Involvement of Uncoupling Protein 2 in Mitochondrial Damage.

Uncoupling protein 2 (UCP2) is essential for maintaining redox homeostasis and regulating energy metabolism. Abnormal expression of UCP2 has been associated with various tumors, including leukemia. Genipin (GEN), a specific inhibitor of UCP2, has a long history of use in traditional Chinese medicine. However, the precise role and underlying mechanisms of UCP2 in the inhibition of leukemia cells by GEN remain inadequately understood. This study focuses on the expression levels of UCP2 in myeloid leukemia (ML) and investigates the effects of GEN on the proliferation, mitochondrial function, and energy metabolism of the chronic myeloid leukemia (CML) cell line K562. The expression of UCP2 in clinical samples and cell lines (HL-60, U937, and K562) was confirmed using real-time quantitative polymerase chain reaction (qPCR) and western blot. The effects of GEN on K562 cell viability, morphology, and apoptosis were assessed through a cell counting kit-8 (CCK-8), Wright-Giemsa staining, and an annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) apoptosis detection kit. Additionally, the impact of GEN on mitochondrial function and energy metabolism, including reactive oxygen species (ROS), mitochondrial membrane permeability transition pore (MPTP), lactic acid (LA), oxygen consumption rate (OCR), and adenosine triphosphate (ATP) levels in K562 cells, was also examined. The results showed that UCP2 was differentially expressed in clinical samples from patients with ML. Among the three cell lines examined, K562 cells exhibited a significantly higher expression level of UCP2. Functionally, GEN markedly inhibited K562 cell viability while promoting K562 cell differentiation and apoptosis. Mechanistically, UCP2 mRNA and protein expression levels were inhibited by GEN in K562 cells in a concentration- and time-dependent manner. Additionally, GEN dramatically increased ROS generation and induced mitochondrial MPTP opening in K562 cells. Furthermore, GEN significantly reduced LA production in K562 cells and markedly increased OCR and ATP production. The results suggest that UCP2 is differentially expressed in ML patients and cell lines; GEN, a UCP2 inhibitor, induces mitochondrial damage and metabolic remodeling, thereby inhibiting proliferation and promoting apoptosis in K562 cells, and thus could be suggested as an adjuvant of an antitumor metabolic therapy.

SMYD3 plays a pivotal role in mediating the epithelial-mesenchymal transition process in breast cancer.

In previous reports, we highlighted the significant involvement of SMYD3, a histone methyltransferase (HMT), in various aspects of cancer progression, including cell adhesion, migration, and invasion. In this study, we delved deeper into understanding the relationship between SMYD3 and epithelial-mesenchymal transition (EMT) both in cell lines and clinical samples. Our investigation uncovered a notable correlation between heightened SMYD3 expression and the presence of EMT markers in human breast cancer tissues. We found that the induction of SMYD3 expression is facilitated by transforming growth factor beta 1 (TGF-β1), which achieves this by suppressing miR-124, an inhibitor that targets SMYD3, through alterations in DNA methylation. Conversely, our experiments demonstrated that reducing SMYD3 levels through RNA interference impeded TGF-β1-induced EMT in breast cancer cells. Furthermore, our results revealed that SMYD3 alone has the capability to modulate the expression of markers associated with EMT. An intriguing aspect of our study is the revelation that SMYD3 influences the activation of vimentin by binding to its response elements within the core promoter region. Notably, this effect is independent of SMYD3's histone methyltransferase activity. These findings collectively underscore the pivotal role of SMYD3 in driving EMT, both in cell lines and primary cancer tissues, particularly emphasizing its significance in TGF-β1-induced EMT in breast cancer.

Identification of RP-54745, an IL-1 Inhibitor Displaying Anticancer Activities for KSHV-Related Primary Effusion Lymphoma.

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of several human cancers, including primary effusion lymphoma (PEL), usually seen in immunocompromised patients while lack of effective therapeutic options. Interleukin-1 (IL-1) family is a major mediator for inflammatory responses and has functional role in both innate and adaptive immunity. We previously showed high activation of multiple IL-1 signaling molecules during KSHV latent and lytic stages, as well as in clinical samples from patients with KSHV-related malignancies. In the current study, we identified RP-54745, a potential antirheumatic compound as IL-1 inhibitor, effectively repressed KSHV + PEL cell growth through inducing tumor cell apoptosis. By using an established PEL xenograft model, we found that RP-54745 treatment suppressed tumor expansion in mice. Also, RP-54745 treatment significantly reduced hyperinflammation in tumor microenvironment including myeloid cells and neutrophils infiltration, as well as blocking IL-1 signaling molecules expression in vivo. In addition, our transcriptome analysis revealed novel cellular genes and mechanisms for anticancer activities of RP-54745. Taken together, our data indicate targeting IL-1 production and signaling may represent promising therapeutic strategies against these virus-associated diseases.

Developing and validating a multiplex hydrolysis probe-based quantitative PCR assay for the detection of four pathogens in chelonians.

Many wildlife conservation efforts focus on the effects of one pathogen, but for many conservation efforts to be successful, researchers require an understanding of ecological processes that may include multiple co-occurring pathogens. We developed a multiplex quantitative PCR (qPCR) assay to detect four pathogens in eastern box turtles (Terrapene carolina carolina), including frog virus 3 (FV3), Terrapene herpesvirus 1 (TerHV1), box turtle Mycoplasma sp. (BTMyco), and Terrapene adenovirus (TerAdv). TaqMan™ primer probes were designed using previously published assays with four different fluorophores. Multiplex Cq values plotted against singleplex Cq values demonstrated slopes of 0.967, 1.00, 0.980, and 0.973 for TerHV1, TerAdv, FV3, and BTMyco, respectively, and R2 values of 0.999 for all four pathogens. The assay was highly consistent with the intra-assay variation of all four pathogen targets, ranging from 0.05-1.826 % across all concentrations, while inter-assay variation ranged from 0.031-4.569 % among all four targets at all concentrations. Clinical samples were tested using previously collected samples from eastern box turtles and red-eared sliders (Trachemys scripta elegans) and performed similarly to singleplex assays. This multiplex assay is an effective, time-efficient diagnostic tool to quickly monitor chelonian pathogens by detecting FV3, TerHV1, BTMyco, and TerAdv within a single reaction. A validated and clinically utilized multiplex assay will be beneficial to characterizing a more complex pathogen profile for future chelonian epidemiological studies to better describe pathogen dynamics and their impacts on individual and population health.

SUMO modified ETV1 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by facilitating CCL2 transcription in esophageal squamous cell carcinoma cells.

Esophageal squamous cell carcinoma (ESCC) is one of the most common malignant tumors with a high metastasis rate and a poor prognosis. ETS variant transcription factor 1 (ETV1) plays an important role in multiple malignancies. However, its function in ESCC progression and tumor microenvironment (TME) remains to be explored. In this study, we characterized the role of ETV1 in ESCC process and TME. Gene expression and immune infiltration in ESCC samples from the Cancer Genome Atlas (TCGA) were analyzed. The expression of ETV1 in clinical samples was detected by real-time PCR, western blot and immunohistochemistry staining. Cell growth was detected by CCK-8 and colony formation assays. Macrophage phenotypes were determined using flow cytometry. Immunofluorescence double staining was used to detect the tumor-associated macrophage (TAM) infiltration. The tumor volume was recorded and weighed. Transcriptional activity was measured using dual-luciferase assay, chromatin immunoprecipitation (ChIP) assay and DNA pull-down assay. In this study, through analysis of ESCC samples from TCGA database and the clinic, we noticed that ETV1 was highly expressed in ESCC tumor tissues and was associated with TAM infiltration. Overexpression of ETV1 promoted ESCC cell proliferation in vitro and xenograft tumor growth in nude mice, while ETV1 knockdown elicited the opposite effects. Furthermore, ETV1 upregulation in tumor tissues was found to drive M2 macrophage infiltration both in vitro (transwell assays) and in vivo (xenograft tumor models). C-C motif chemokine ligand 2 (CCL2), a key factor inducing M2 macrophage polarization, was also found to be elevated in ESCC tumor tissues. Mechanism study demonstrated that ETV1 facilitated M2 macrophage infiltration via the transcriptional modulation of CCL2. In addition, the cause of the changes in ETV1 activity and expression was investigated. The E2 small ubiquitin-like modifier (SUMO) binding enzyme UBC9 increased ETV1 activity and expression, indicating the presence of SUMO modification in ETV1. Our data deciphered the mechanism of ETV1-mediated M2 macrophage infiltration in the TME of ESCC, which has important implications for the development of novel prognostic and therapeutic targets to optimize current therapies against ESCC.

Development of a multiplex droplet digital PCR method for detection and differentiation of mpox virus clades

BackgroundThe current outbreak of mpox has been declared a public health emergency of international concern by the World Health Organization. However, distinguishing symptoms of mpox virus (MPXV) infection from other orthopoxviruses is atypical, necessitating laboratory confirmatory tests to aid in clinical diagnosis. Therefore, rapid and accurate detection and differentiation of various clades of MPXV are imperative. ObjectiveA multiplex droplet digital PCR (ddPCR) method was developed to detect and differentiate various clades of MPXV with subsequent evaluation of its sensitivity and accessibility through the analysis of 17 clinical samples. MethodsPrimers and probes for multiple ddPCR were designed by comparing multiple complete genomes of orthopoxviruses. Primer and probe concentrations, reaction conditions were tentatively optimized on the Biorad QX200 platform. Seventeen clinical samples of MPXV were detected and verified by Sanger sequencing. ResultsThe established ddPCR method could detect and differentiate MPXV, and the results were consistent with those of Sanger sequencing. ConclusionMultiplex ddPCR could be used to detect and distinguish different clades of MPXV rapidly and accurately.

Performance evaluation of TaqMan™ Arbovirus Triplex Kit (ZIKV/DENV/CHIKV) for detection and differentiation of Dengue and Chikungunya viral RNA in serum samples of symptomatic patients

IntroductionGlobal outbreaks of mosquito-transmitted arbovirus infections, such as dengue (DENV) and chikungunya (CHIKV), are increasing. Differentiating these infections is challenging due to non-specific symptoms and serology limitations. PCR-based approaches offer higher sensitivity and specificity. This study evaluated the performance of TaqMan™ Arbovirus Triplex Kit (ZIKV/DENV/CHIKV) (TaqMan™ Kit) to detect DENV and CHIKV in clinical samples from patients in south India. MethodsIn total, 280 serum samples with 90 DENV-positive, 90 CHIKV-positive, and 100 negative samples were tested with TaqMan™ Kit and CDC Trioplex Real-Time RT-PCR assay. No Zika virus was detected. The sensitivity and specificity of viral RNA detection were determined, and discordant results were resolved using comparator PCRs, dengue NS1 antigen detection, virus-specific antibody results, or previously de-identified in-house PCR results. ResultsTaqMan™ Kit showed 100% agreement with the comparator for DENV detection in 92 positive samples. Among 188 samples negative for DENV by the comparator, 30 showed positive results with the TaqMan™ kit, and 23 of those were confirmed as true positives. The resulting sensitivity and specificity for DENV detection were 100% and 95.1%, respectively. For CHIKV, 77 positive and 195 negative results were concordant. Eight samples showed discordant results, but upon resolution testing, sensitivity and specificity for CHIKV were 93.9% and 100.0%, respectively. ConclusionTaqMan™ Arbovirus Triplex Kit demonstrated high sensitivity and specificity (>93%) for detecting circulating DENV and CHIKV strains. Multiplex PCR testing can improve case detection, surveillance, and public health responses while optimizing laboratory resources for outbreak control.

Comprehensive analysis of chromosome abnormalities by chromosome conformation based karyotyping (C-MoKa) in patients with conception failure and pregnancy loss.

Chromosome abnormalities are a leading cause of conception failure and pregnancy loss. While traditional cytogenetics technologies like karyotyping have been helpful in identifying structural variations (SVs), they face challenges in detecting complex rearrangements and cryptic structures. In this study, we developed a new method called chromosome conformation based karyotyping (C-MoKa) to comprehensively detect different types of chromosomal abnormalities in patients with conception failure and pregnancy loss. A total of 70 clinical samples exhibiting known results of SVs, mosaic aneuploidies, copy number variations （CNVs） and uniparental disomy (UPD) were included in our cohort and underwent C-MoKa analysis. The results obtained from different techniques, including karyotyping, CNV-seq, and CMA were compared and analyzed. Distinct chromosomal conformation patterns of various variations were observed and analyzed in clinical samples. Our C-MoKa method not only validated all the findings of karyotyping, CNV-seq and CMA, but also provided more detailed results. It demonstrated superior fragment resolution (<500Kb) and more precise breakpoints (>100kb). Moreover, C-MoKa showed higher sensitivity in decoding intricate rearrangements in a single test. Our results highlight the potential utility of C-MoKa in precisely unraveling SVs, mosaic aneuploidies, CNVs, and UPD in clinical settings, which can significantly impact further clinical decision-making.

Development of a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Beiji nairovirus.

Beiji nairovirus (BJNV) is a recently discovered tick-borne RNA virus associated with human febrile illness. This study aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the precise detection of BJNV, with a specific focus on assessing its effectiveness with clinical samples. The optimal molecular target was identified as the BJNV small (S) segment gene, and the ideal reaction conditions were established at 65 °C for 50 min. A neutral red stain concentration of 300 μM was determined to be optimal for visualizing the LAMP reaction. The LAMP method demonstrated an impressive lower limit of detection at 10 copies/μL, highlighting a sensitivity level 10,000 times higher than traditional PCR methods. Moreover, it showed no cross-reactivity with three other viruses. The LAMP assay demonstrated results consistent with those of semi-nested PCR when applied to clinical samples, thereby validating its suitability for field testing. In conclusion, the LAMP assay developed in this study represents a significant advancement in the rapid detection of BJNV. Its high sensitivity, specificity, and ease of use make it a valuable tool for the establishment of effective prevention and control strategies against this emerging pathogen.

Effects of targeting CTMP on immunogenicity of MSI-L colon adenocarcinoma cells by metabolism and cell cycle modulation.

246 Background: Regimen of metastatic colon cancer (mCOAD) is moving towards combination of immune checkpoint inhibitors (ICBs) and targeted therapy, however ICBs are rarely effective in MSI-L metastatic colon cancers, limiting options for backline treatments, new targeted therapy is on demand to improve immunogenicity of MSI-L mCOAD. As targeting fatty acid metabolism or cell cycle has shown potential in improving the efficacy of ICBs, our team focused on CTMP, a thioesterase (ACOT) family member, mainly modulates fatty acid β oxidation, meanwhile it binds to phosphorylation sites of AKT thus intervenes cell cycle, currently its role on immunogenicity of COAD is not clear. Methods: Correlation between CTMP and OS in MSI-L colon cancer patients was assessed by KM plotter in GEO database.11 Clinical samples of MSI-L colon cancer, who received a third-line treatment regimen of anti-PD-1 and fruquintinib, were collected then applied in immunofluorescence and western blot analyses to evaluate CTMP and MHC-I expression. In vitro, Lentiviral transfection of targeted genes were stably established,then applied in RNA-seq and metabolic assays. Proteomic analysis and co-IP were applied to explore CTMP-modulating pathway. In vitro, IFNγ was intratumorally injected. Results: KM curve showed CTMP was negatively correlated with OS in MSI-L COAD in GEO database. In clinical samples, lower CTMP expression and higher MHC-I expression were observed in partial response (PR) than progressive disease (PD) group. In vitro, CTMP-OE reduced MHC-I, increased AKT phosphorylation and immune checkpoints (PD-L1, IDO-1), promoted clone formation. Seahorse assay showed increased capacity of glycolysis and oxidative phosphorylation in CTMP-OE than vector. RNA-seq showed CTMP-OE reduced MHC-I augmentation under IFN-γ treatment, meanwhile glucose metabolism genes, ACOT family ,immune checkpoints were also up-regulated, indicating ACOT family related metabolism promotion and immunogenicity suppression of COAD. Proteomic analysis and co-IP assay indicated CTMP interacts REV7, a key mitotic regulatory protein. Enzyme activity assay showed REV7 negatively regulates CDK1. Furthermore, REV7-OE alone is capable to induce MHC I augmentation and G2 arrest, accordingly CTMP-KD enhanced REV7's negative modulation on CDK1,contributing to G2 arrest, that also augmented MHC I . In homologous mice tumor-bearing model, tumor shrinkage was observed in CTMP-KD/ REV7-OE group. CTMP-KD/ REV7-OE increases MHC I expression,CD8+ T cell infiltration under IFN γ intra-tumoral injection. Conclusions: In summary, our studies suggested that CTMP regulates the metabolism of colorectal cancer and intervenes MHC-I molecules expression, and targeting CTMP is potential in suppressing tumor cell cycle and promoting immunogenicity through its modulation on REV7.

Benefits of the “worst-case scenario”: a multi-level examination of the effects of confronting the feared outcome during imagery-based exposure

ABSTRACT Exposure therapy is an efficacious treatment for anxiety-related disorders. Yet, fear often returns after treatment. Occasional reinforcement, in which the feared stimulus is intermittently presented during extinction, increases safety learning and slows fear renewal in conditioning paradigms and analogue samples, but no studies to date have examined this strategy in clinical samples. The present study examined the effects of vicarious occasional reinforcement on fear renewal in a snake-phobic sample across multiple levels of analysis. Fear was intermittently reinforced by providing reminders of the feared outcome (a snake bite) throughout a two-session analogue video exposure manipulation. Snake-phobic adults were randomized to one of three conditions: a single-cue [S], multiple-cue [M], or multiple-cue+fear-outcome [M+FO] exposure group. Results showed the three groups did not significantly differ in threat expectancy or attentional bias for threat at follow-up. Despite sustained anxiety, however, the M+FO condition completed significantly more steps on a visual avoidance task at follow-up than the M and S conditions and heightened mean distress during exposure mediated this effect. The M and S groups did not significantly differ in visual avoidance at follow-up. These findings suggest incorporating reminders of the feared outcome into exposure may be an effective strategy for increasing inhibitory retrieval.

IGSF8 impairs migration and invasion of trophoblast cells and angiogenesis in preeclampsia.

Insufficient trophoblast cell infiltration is implicated in the progression of preeclampsia (PE). The immunoglobulin superfamily member 8 (IGSF8) has been shown to promote cell migration, invasion, and epithelial mesenchymal transition (EMT). However, the specific impact of IGSF8 on trophoblast cells in PE has not been definitively demonstrated. To address this, placental tissues from PE patients and normal subjects was collected. A PE-like rat model was established by administering L-NAME (60mg/kg) intragastrically to pregnant rats from the 10th to the 19th day of gestation. Knockdown and overexpression plasmids of IGSF8 were transfected into JEG-3cells for further experiments. Clinical samples indicated impaired spiral artery remodeling, and high IGSF8 expression in the placental tissues of PE patients. PE rats exhibited increased mean arterial pressure, elevated 24-h urine protein levels, higher abortion rates, and decreased placental and fetal weight compared to rats of sham group. Failure of physiological transformation of spiral arteries was observed in PE rats, along with increased IGSF8 expression. IGSF8 overexpression inhibited JEG-3cell migration, invasion and EMT, as well as reduced release of VEGF in JEG-3cells, impairing HUVEC tube formation. mRNA-sequencing analysis of JEG-3cells transfected with shIGSF8 showed differentially expressed genes related to angiogenesis, and mesenchymal cell differentiation, with IGSF8 knockdown being associated with the activation of pathways involved in blood vessel development and cell migration. Overall, this study suggests that IGSF8 plays a role in the development of PE and provides new insights for potential treatments.

Natural history study of patients with pancreatic acinar cell carcinoma: A 3-year prospective longitudinal analysis.

764 Background: Pancreatic acinar cell carcinoma (PACC) is a rare exocrine tumor of the pancreas that comprises 0.2-2% of all pancreatic malignancies. There are no prospective randomized studies in this disease and most data about PACC comes from retrospective database analyses and case reports. Methods: The Cancer Moonshot-funded My Pediatric and Adult Rare Tumor (MyPART) network’s Rare Solid Tumor Natural History study (NCT03739827) is a prospective, single-institution observational study that enrolls patients of all ages with rare solid tumors (&lt;15 cases per 100,000 people per year), including PACC. Patients can participate remotely (field cohort) or visit the NIH for annual evaluations (in-person cohort). All participants complete medical and family histories, patient reported outcomes (PROs) measures, and provide saliva for DNA analysis. Relevant data such as clinical and family histories, surgical and molecular pathology, and imaging results are extracted from patient medical records and entered in the central study database. Archival tumor samples (when available) are analyzed using a 500+ gene panel (TruSight500, Illumina) and discussed in a molecular tumor board. Patients participating in the in-person cohort undergo additional evaluations including clinical examination, imaging studies, genetic counseling, and blood sample collection for clinical and research purposes (standard clinical labs, germline DNA/RNA, immune phenotypes, cytokines) as indicated. Results: From 09/2021 until 09/2024, 17 PACC patients (n=13 for the field cohort, n=4 for the in-person cohort) were accrued. The mean age of diagnosis is 64.7 (48-78) years, and the male to female ratio is 14:3. At the time of diagnosis, 6 patients had Stage I or II, 1 had Stage III, and 10 had Stage IV disease. The median follow-up time since accrual is 7.6 months, and 4 patients have died. No patients have been lost to follow-up. Of the 6 patients diagnosed with early-stage disease, 5 had surgery and all received adjuvant chemotherapy, while 1 patient is receiving neoadjuvant chemotherapy. Among the 9 patients diagnosed with advanced disease, 8 received platinum-based regimens as a first-line therapy. Tumor molecular profiling was available for 16 patients. The most common tumor mutations observed were in CDKN2A (29.4%), BRCA2 (29.4%), and SMAD4 (23.5%). Six patients received targeted therapies matched to their tumor molecular profile. Conclusions: The demographic profile of our cohort aligns with previous studies, with a mean age of diagnosis in the early-to-mid 60s and a high male to female ratio. Our cohort highlights a shift in first-line treatment to platinum-based therapies over the past decade. Many PACC tumors have targetable mutations found in molecular profiling, and the efficacy of matched therapy warrants further exploration. Clinical trial information: NCT03739827 .

Serologic diagnosis of Entamoeba histolytica infection based on the gradient-based digital immunoassay.

Entamoeba histolytica is a parasite that could cause severe amebiasis, an extremely contagious parasitic disease with critical clinical symptoms. Timely diagnosis and treatment of E. histolytica are crucial for preventing complications and fatalities. However, current methods for E. histolytica diagnosis still rely on traditional and laborious methods such as stool examinations and serological immunoassay. Those methods urgently need improved diagnostic sensitivity, diagnostic specificity, detection time, and diagnostic accuracy. Herein, we developed a rapid and accurate gradient-based digital immunoassay system for diagnosing E. histolytica. This method uses a homemade pre-coated recombinant Igl-C fragment with high binding affinity, designed, recombinantly expressed, and purified by our lab on a one-shot observation microfluidic chip to capture the disease biomarker, specific anti-Igl-C antibodies in the serum. The captured antibodies are detected by anti-human IgG-coated gold nanoparticles, forming a digital binding sandwich immunocomplex. After simple imaging processing, the number of gold nanoparticles bound to the reaction surface within the same field of view of the microscope image was analyzed. Then, it can be easily determined in about 15min whether specific anti-Igl-C antibodies are in the serum, allowing quicker identification of E. histolytica infection. The detection and diagnosis performance were confirmed by testing 25 clinical serum samples. The novel diagnostic method provided a digital immunoassay platform and has a strong potential to be a high-efficiency tool for the clinical diagnosis of E. histolytica infection with heightened diagnostic sensitivity and diagnostic specificity. It offers the potential for rapid point-of-care testing, facilitating prompt treatment initiation and improved patient outcomes in resource-limited healthcare settings, making it a valuable asset in the fight against amebiasis.

UGCG promotes chemoresistance and breast cancer progression via NF-κB and Wnt/β-catenin pathway activation.

Taxane-based chemotherapy is the primary treatment for triple-negative breast cancer (TNBC), yet clinical outcomes remain unsatisfactory due to the persistence of chemoresistance. Identifying key factors that contribute to chemoresistance and understanding the associated molecular mechanisms is therefore essential. The GEO databases were utilized to pinpoint factors related to chemoresistance, which were subsequently validated using clinical tissue samples. The role of UGCG in the malignant progression and chemoresistance of TNBC was assessed through various functional assays. Western blotting, qRT-PCR, and immunohistochemistry were employed to investigate the signaling pathways associated with UGCG in TNBC. UGCG expression was notably elevated in chemoresistant breast cancer tissues and cells, as identified in GEO databases and confirmed through immunohistochemistry. Additionally, findings from our cohorts indicated that higher levels of UGCG expression correlated with a lower rate of pathological complete response (pCR), suggesting it could serve as an independent predictor of chemotherapy effectiveness. Gain- and loss-of-function experiments demonstrated that UGCG enhanced the proliferation, metastasis, and stemness of breast cancer cells. Furthermore, treatment with paclitaxel or docetaxel resulted in increased UGCG expression, which in turn reduced chemotherapy-induced cell apoptosis and improved drug resistance and metastatic capabilities. Mechanistically, UGCG was found to amplify the activation of NF-κB and Wnt/β-catenin pathways, and the use of inhibitors targeting these pathways diminished the UGCG-induced malignant effects. Our findings underscore the significant role of UGCG in the chemoresistance and progression of breast cancer, suggesting it as a predictive biomarker and potential therapeutic target to combat chemoresistance in this disease.

Regulation of pancreatic cancer cells by suppressing KIN17 through the PI3K/AKT/mTOR signaling pathway.

Pancreatic cancer is an aggressive tumor, which is often associated with a poor clinical prognosis and resistance to conventional chemotherapy. Therefore, there is a need to identify new therapeutic markers for pancreatic cancer. Although KIN17 is a highly expressed DNA‑ and RNA‑binding protein in a number of types of human cancer, its role in pancreatic cancer development, especially in relation to progression, is currently unknown. The present study verified the upregulation of KIN17 in pancreatic cancer using The Cancer Genome Atlas and Gene Expression Omnibus databases (GSE15471, GSE71989 and GSE62165), and identified an association between the PI3K/Akt/mTOR pathway and patient prognosis using publicly available datasets (Gene Expression Profiling Interactive Analysis). Immunohistochemistry was performed to determine the association between KIN17 and the pathological features of clinical pancreatic cancer samples. Furthermore, knockdown of KIN17 was shown to inhibit the migration and invasion of pancreatic cancer cells, and to reverse epithelial‑mesenchymal transition in pancreatic cancer cells through downregulation of Vimentin and N‑cadherin, and upregulation of E‑cadherin. Through various cellular experiments, the role of KIIN17 was explored in PI3K/AKT/mTOR activity. KIN17 inhibition was shown to suppress the migration and invasion of pancreatic cancer cells through PI3K/AKT/mTOR‑mediated autophagy. Furthermore, combined with mTOR inhibition, dual inhibition could enhance autophagy, leading to anti‑migratory and anti‑invasion effects in pancreatic cancer. In conclusion, the present study indicated that KIN17 may have a role in carcinogenesis and could serve as a prognostic biomarker of pancreatic cancer, owing to its high expression. In addition, KIN17 may be considered a potential therapeutic target with its knockdown having an inhibitory effect on pancreatic cancer.