Translatome analysis in acute ischemic stroke: Astrocytes and microglia exhibit differences in poststroke alternative splicing of expressed transcripts.

Astrocytes and microglia undergo dynamic and complex morphological and functional changes following ischemic stroke, which are instrumental in both inflammatory responses and neural repair. While gene expression alterations poststroke have been extensively studied, investigations into posttranscriptional regulatory mechanisms, specifically alternative splicing (AS), remain limited. Utilizing previously reported Ribo-Tag-seq data, this study analyzed AS alterations in poststroke astrocytes and microglia from young adult male and female mice. Our findings reveal that in astrocytes, compared to the sham group, 109 differential alternative splicing (DAS) events were observed at 4 h poststroke, which increased to 320 at day 3. In microglia, these numbers were 316 and 266, respectively. Interestingly, the disparity between DAS genes and differentially expressed genes is substantial, with fewer than 10 genes shared at both poststroke time points in astrocytes and microglia. Gene ontology enrichment analysis revealed the involvement of these DAS genes in diverse functions, encompassing immune response (Adam8, Ccr1), metabolism (Acsl6, Pcyt2, Myo5a), and developmental cell growth (App), among others. Selective DAS events were further validated by semiquantitative RT-PCR. Overall, this study comprehensively describes the AS alterations in astrocytes and microglia during the hyperacute and acute phases of ischemic stroke and underscores the significance of certain hub DAS events in neuroinflammatory processes.

Evaluation of the diagnostic performance of colposcopy in the detection of cervical high-grade squamous intraepithelial lesions among women with transformation zone type 3.

BACKGROUND: Inaccurate colposcopy diagnosis may lead to inappropriate management and increase the incidence of cervical cancer. This study aimed to evaluate the diagnostic accuracy of colposcopy in the detection of histologic cervical intraepithelial neoplasia grade 2 or worse (CIN2+) in women with transformation zone type 3 (TZ3). METHODS: Records from 764 patients with TZ3 who underwent colposcopy-directed biopsy and/or endocervical curettage in Putuo Hospital China between February 2020 and March 2023 were retrospectively collected. Colposcopy was carried out based on 2011 International Federation of Cervical Pathology and Colposcopy (IFCPC) and Colposcopy nomenclature. The diagnostic performance of colposcopy for identifying CIN2 + was evaluated compared with biopsies. The Kappa and McNemar tests were used to perform statistical analyses. RESULTS: Among the study population, 11.0% had pathologic CIN2+. The relative sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of colposcopy for histologic CIN2 + were 51.2%, 96.5%, 64.2% and 94.1%, respectively. The senior colposcopists (80.6%) had a higher colposcopic accuracy to diagnose histologic CIN2 + than junior colposcopists (68.6%). In subgroup analyses, age group ≥ 60 years (70.3%) showed lowest diagnostic accuracy when compared with age groups of < 45 years (84.4%) and 45-59 years (74.9%). CONCLUSION: Our findings suggest an increased risk of diagnostic inaccuracy of colposcopy in identifying CIN2 + in those ≥ 60 years of age with TZ3, and the accuracy of colposcopy is required to be further improved.

An electrochemical biosensor based on DNA tetrahedron nanoprobe for sensitive and selective detection of doxorubicin.

Doxorubicin (DOX) is a clinical chemotherapeutic drug and patients usually suffer from dose-dependent cytotoxic and side effects during chemotherapy process with DOX. Therefore, developing a reliable strategy for DOX analysis in biological samples for dosage guidance during chemotherapy process is of great significance. Herein, a sensitive and selective electrochemical biosensor for DOX detection was designed based on gold nanoparticles (AuNPs) and DNA tetrahedron (TDN) nanoprobe bifunctional glassy carbon electrode that could detect DOX in human serum and cell lysate samples. AuNPs not only could enhance electron transfer efficiency and detection sensitivity, but also could improve the biocompatibility of electrode. TDN nanoprobes were employed as specific DOX bind sites that could bind abundant DOX through intercalative characteristics to contribute to sensitive and selective detection. Under the optimal conditions, the proposed TDN nanoprobes-based DOX biosensor exhibited a wide linear range that ranged from 1.0 nM to 50 µM and a low detection limit that was 0.3 nM. Moreover, the proposed DOX biosensor displayed nice selectivity, reproducibility and stability, and was successfully applied for DOX detection in human serum and cell lysate samples. These promising results maybe pave a way for DOX dosage guidance and therapeutic efficacy optimization in clinic.

Detection of cervical high-grade squamous intraepithelial lesions and assessing diagnostic performance of colposcopy among women with oncogenic HPV.

BACKGROUND: HPV screening tests may improve cervical cancer risk stratification and better guide decisions about follow-up with colposcopy/biopsy. This study aimed to estimate the risk of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) among women with oncogenic HPV types and evaluate the performance of colposcopy in the diagnosis of histologic CIN2 + at Putuo Hospital, Shanghai, China. METHODS: This cross-sectional survey was conducted from February 2020 to December 2022 among women who were referred to colposcopy. Women with high-risk (HR) HPV-positive, cytology testing and colposcopy-directed biopsy were included. RESULTS: Univariate and multivariate analysis indicated that high-grade colposcopic impression ((OR, 17.61%, 95%CI: 11.54-26.85%) was associated with the highest risk for detecting CIN2+, followed by HSIL + cytology (OR, 6.90%, 95%CI: 3.56-13.37%) and HPV16/18 positive (OR, 2.91%, 95%CI: 2.12-3.99%). Overall, CIN2 + was detected in 14.6% of 2007 women. HPV16/18 had higher CIN2 + risks than other HR-HPV genotypes (30.1% vs. 10.2%, P<0.001). Among women with low-grade cytology, 24.1% had CIN2+, and the risks for HPV16/18 (58.2%) were higher than for other HR-HPV(16.8%). For those with high-grade cytology, there was no significant difference between HPV groups ( 75.0% vs. 72.9%, P > 0.05). The diagnostic performance of colposcopy in diagnosis of CIN2 + by senior and junior colposcopists was comparable. CONCLUSIONS: The results indicated that referral to colposcopy is recommended in managing women with HR-HPV positive, and colposcopic impressions provide key clues for identification CIN2+.

Glutamate-aspartate transporter 1 attenuates oxygen-glucose deprivation-induced injury by promoting glutamate metabolism in primary cortical neurons.

Ischemic stroke is a common cerebral disease. However, the treatment for the disease is limited. Daurian ground squirrel (GS; Spermophilus dauricus), a hibernating mammalian species, is highly tolerant to ischemia. In the present study, GS neurons in a non-hibernating state were found to be more resistant to oxygen-glucose deprivation (OGD), an ischemic model in vitro. We leveraged the differences in the endurance capacity of GS and rats to investigate the mechanisms of resistance to ischemia in GS neurons. We first identified glutamate-aspartate transporter 1 (GLAST) as a cytoprotective factor that contributed to tolerance against OGD injury of GS neurons. The expression of GLAST in GS neurons was much higher than that in rat neurons. Overexpression of GLAST rescued viability in rat neurons, and GS neurons exhibited decreased viability following GLAST knockdown under OGD conditions. Mechanistically, more glutamate was transported into neurons after GLAST overexpression and served as substrates for ATP production. Furthermore, eukaryotic transcription initiation factor 4E binding protein 1 was downregulated by GLAST to rescue neuronal viability. Our findings not only revealed an important molecular mechanism underlying the survival of hibernating mammals but also suggested that neuronal GLAST may be a potential target for ischemic stroke therapy.

Analysis of risk factors of non-suicidal self-harm behavior in adolescents with depression.

BACKGROUND: Adolescents are a high-risk group for non-suicidal self-injury (NSSI), which seriously affects their physical and mental health. This study aimed to explore the risk factors for depressive adolescents with NSSI. METHODS: A total of 153 adolescents with depression were divided into the NSSI group (n=65) and non-NSSI group (n=88) according to the criteria stipulated by Diagnostic and Statistical Manual of Mental Disorders, 5th Edition (DSM-5). The Beck scale for suicidal ideation (BSS), adolescent self-rating life events checklist (ASLEC), family adaptability and cohesion evaluation scale II-Chinese version (FACES II-CV), childhood trauma questionnaire short form (CTQ SF), and multidimensional scale of perceived social support (MSPSS) were applied to evaluate suicidal ideation, frequency and intensity of stressful life events, family functions, childhood trauma, and perceived support, respectively. We applied two-dimensional logistic regression to identify risk factors for NSSI. RESULTS: Female gender ratio, suicidal ideation, and attempted suicide were significantly higher in the NSSI group than in the non-NSSI group (all P<0.05). Scores of interpersonal relationships in ASLEC, emotional abuse, and emotional neglect in the CTQ-SF were significantly higher in the NSSI group than those in the non-NSSI group (all P<0.001). The scores of family cohesion (P=0.001) and family adaptability (P=0.01) were significantly lower in the NSSI group than in the non-NSSI group. The MSPSS was used to assess support from the family, and the index was significantly lower in the NSSI group (P<0.001). After adjusting for age and gender, BSS score, interpersonal relationship score, emotional abuse score, and emotional neglect score were identified as independent risk factors for NSSI. CONCLUSIONS: The rate of NSSI in adolescents with depression is high. Higher scores of BSS, interpersonal relationship, emotional abuse, and neglect were independently associated with NSSI.

Hydrogen Sulfide Contributes to Uterine Quiescence Through Inhibition of NLRP3 Inflammasome Activation by Suppressing the TLR4/NF-κB Signalling Pathway.

BACKGROUND: The NLRP3 inflammasome plays a critical role in inflammatory responses in various diseases. Our previous study showed that NLRP3 expression was significantly increased in human pregnancy tissue during term labour. Therefore, we explored whether NLRP3 participated in inflammatory responses of preterm and term labour and whether this process could be relieved by H2S, one anti-inflammatory gasotransmitter. METHODS: Human myometrium was obtained from non-labouring and labouring women. Mouse myometrium was obtained from LPS-induced infectious preterm labour. Uterine smooth muscle cells were isolated from non-labouring women's myometrial tissues, transfected with siRNA, and treated cells with IL-1ß, H2S donor NaHS, NF-κB inhibitor BAY 11-7082 and TLR4 inhibitorTAK-242. The NLRP3 inflammasome, CSE, CBS, TLR4, uterine contraction-associated proteins (CAPs), NF-κB activation and inflammatory cytokine expression were assessed by Western blotting and RT-PCR. RESULTS: The NLRP3 inflammasome, TLR4 and activated NF-κB expression were upregulated in human term labour, mouse preterm labour and human uterine smooth muscle cells treated with IL-1ß. NLRP3 levels were negatively correlated with CSE and CBS expression. Treatment with the H2S donor NaHS delayed LPS-induced preterm birth in mice and inhibited NLRP3 inflammasome activation. In siNLRP3-transfected cells, there was a significant decrease in the expression of CAPs and inflammatory cytokines compared with IL-1ß stimulation. In addition, treatment with the H2S donor NaHS inhibited NLRP3 inflammasome activation, reduced the expression of uterine contraction-associated proteins and inflammatory cytokines and reduced the activation of TLR4 and NF-κB compared with stimulation with IL-1ß in human uterine smooth muscle cells. Furthermore, treatment of uterine smooth muscle cells with BAY 11-7082 and TAK-242 found that NLRP3 activation was regulated by the TLR4 and NF-κB pathways. CONCLUSION: H2S suppresses CAP expression and the inflammatory response and contributes to uterine quiescence by inhibiting the TLR4/NF-κB signalling pathway and downstream NLRP3 inflammasome activation. Thus, H2S contributes to uterine quiescence through inhibition of NLRP3 inflammasome activation by suppressing the TLR4/NF-κB signalling pathway.

Is cognitive behavioral therapy a better choice for women with postnatal depression? A systematic review and meta-analysis.

The present study evaluated the combined effectiveness of cognitive behavioral therapy (CBT) for postnatal depression. A systematic search was conducted across databases including PubMed, Embase, and the Cochrane library to identify the randomized controlled trials (RCTs) that assessing CBT versus control for postnatal depression until March 2017. Data was extracted by two reviewers, independently. The Review Manager 5.3 and Stata 11.0 were used to calculate the synthesized effect of CBT on depression, and anxiety. A total of 20 RCTs involving 3623 participants were included. The results of meta-analysis showed that CBT was associated with a better Edinburgh Postnatal Depression Scale (EPDS) than control in short-term (mean difference = -2.86, 95% CI: -4.41--1.31; P<0.05) and long-term (mean difference = -1.68, 95% CI: -1.81-1.56; P<0.05). CBT also improved short-term (mean difference = -6.30, 95% CI: -11.32--1.28; P<0.05) and long-term (mean difference = -4.31, 95% CI: -6.92--1.70; P<0.05) Beck Depression Inventory (BDI). Subgroup analysis based on intervention types showed that in-home and telephone-based therapy exhibited significant reductions in EPDS scores (P<0.05 for all). CBT significantly improved the short-term [odds ratio (OR) = 6.57, 95% CI: 1.84-23.48; P<0.05] and long-term (OR = 2.00, 95% CI: 1.61-2.48; P<0.05) depressive symptomatology as compared to control. CBT also reduced the score of Depression Anxiety Stress Scales (DASS), though without significance. In conclusion, CBT effectively improved the symptoms and progression of postnatal depression.

The Inhibitory Effect of α/ß-Hydrolase Domain-Containing 6 (ABHD6) on the Surface Targeting of GluA2- and GluA3-Containing AMPA Receptors.

The α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors (AMPARs) are major excitatory receptors that mediate fast neurotransmission in the mammalian brain. The surface expression of functional AMPARs is crucial for synaptic transmission and plasticity. AMPAR auxiliary subunits control the biosynthesis, membrane trafficking, and synaptic targeting of AMPARs. Our previous report showed that α/ß-hydrolase domain-containing 6 (ABHD6), an auxiliary subunit for AMPARs, suppresses the membrane delivery and function of GluA1-containing receptors in both heterologous cells and neurons. However, it remained unclear whether ABHD6 affects the membrane trafficking of glutamate receptor subunits, GluA2 and GluA3. Here, we examine the effects of ABHD6 overexpression in HEK293T cells expressing GluA1, GluA2, GluA3, and stargazin, either alone or in combination. The results show that ABHD6 suppresses the glutamate-induced currents and the membrane expression of AMPARs when expressing GluA2 or GluA3 in the HEK293T cells. We generated a series of GluA2 and GluA3 C-terminal deletion constructs and confirm that the C-terminus of GluAs is required for ABHD6's inhibitory effects on glutamate-induced currents and surface expression of GluAs. Meanwhile, our pull-down experiments reveal that ABHD6 binds to GluA1-3, and deletion of the C-terminal domain of GluAs abolishes this binding. These findings demonstrate that ABHD6 inhibits the AMPAR-mediated currents and its surface expression, independent of the type of AMPAR subunits, and this inhibitor's effects are mediated through the binding with the GluAs C-terminal regions.

[Determination of 14 aniline derivatives in water by liquid chromatography-tandem mass spectrometry].

A new and fast method was developed for the simultaneous determination of 14 aniline derivatives ( ADs) in water by direct injection-liquid chromatography-tandem mass spectrometry (LC-MS/MS) through optimizing chromatographic and mass spectrometric conditions. The water sample was filtered through a 0.45 µm polyether sulfone (PES) microfiltration membrane. The separation was performed on a Shim-pack FC-ODS column (75 mm x 4.6 mm, 3 µm) with methanol-0.1% (v/v) formic acid aqueous solution (35:65, v/v) as mobile phases in gradient elution mode. The flow rate was 0.3 mL/min, and the column temperature was 35 °C. The analytes were detected by LC-MS/MS in multiple reaction monitoring mode. Under the optimized conditions, the analysis of the 14 aniline derivatives was completed within 12 min and the calibration curves showed good linearity with correlation coefficients not less than 0.999. The detection limits of the 14 aniline derivatives ranged from 0.03 µg/L to 4. 19 µg/L. The relative standard deviations of the 14 aniline derivatives in the spiked surface water at three levels (0. 5, 5.0, 20.0 µg/L) were 0.4%-9.4% (n = 6). The proposed method has the advantages of good anti-interference ability, rapidness and high sensitivity. It was successfully applied to the analysis of real samples, and the recoveries of the 14 aniline derivatives in the spiked real samples were 68.0%-130%.

Application of GC-triple quadrupole MS in the quantitative confirmation of polycyclic aromatic hydrocarbons and phthalic acid esters in soil.

A new multi-residue method has been developed and validated for the simultaneous analysis of 34 polycyclic aromatic hydrocarbons (PAHs) and phthalic acid esters (PAEs) in soil at trace levels by gas chromatography coupled to triple quadrupole mass spectrometry. Microwave extraction and solid-phase extraction have been employed prior to gas chromatography tandem mass spectrometry analysis. Quality parameters have been established using matrix spike and reference material IRM 104A. Average recoveries of the 34 organic compounds spiked at 5 microg/kg into soils are typically in the range of 66.59-122.07% with relative standard deviations generally less than 20%. Limits of detection (LODs) for PAEs are < or = 0.84 microg/kg, and limits of quantification (LOQs) ranged from 0.13 to 2.81 microg/kg. LODs for PAHs are < or = 0.51 microg/kg, and LOQs ranged from 0.02 to 1.81 microg/kg.

Sensitive determination of aliphatic amines by high-performance liquid chromatography with a new fluorogenic probe 3-(4-fluorinebenzoyl)-2-quinoline carboxaldehyde.

A new fluorogenic reagent 3-(4-fluorinebenzoyl)-2-quinoline carboxaldehyde (FBQCA) has been synthesized and used as a derivatizing reagent for the determination of aliphatic amines with HPLC. The reagent is nonfluorescent, but forms highly fluorescent isoindole upon the reaction with primary amines in alkaline medium. Eleven amine derivatives were baseline separated in 8 min using a gradient elution on a C(8) column and detected with fluorescence detection at lambda(ex)/lambda(em) = 480/546 nm. The detection limits were in the range of 0.5-2 nM (S/N = 3). The proposed method has been successfully applied to the analysis of aliphatic amines in food and environmental samples, including white wine, soybean oil, soil, and tap water with satisfactory recoveries in the range of 94-106%.