Dynamics of N6-methyladenosine modification during Alzheimer's disease development.

N6-methyladenosine (m6A) modification is a common RNA modification in the central nervous system and has been linked to various neurological disorders, including Alzheimer's disease (AD). However, the dynamic of mRNA m6A modification and m6A enzymes during the development of AD are not well understood. Therefore, this study examined the expression profiles of m6A and its enzymes in the development of AD. The results showed that changes in the expression levels of m6A regulatory factors occur in the early stages of AD, indicating a potential role for m6A modification in the onset of the disease. Additionally, the analysis of mRNA m6A expression profiles using m6A-seq revealed significant differences in m6A modification between AD and control brains. The genes with differential methylation were found to be enriched in GO and KEGG terms related to processes such as inflammation response, immune system processes. And the differently expressed genes (DEGs) are negatively lryassociated with genes involved in microglia hemostasis, but positively associated with genes related to "disease-associated microglia" (DAM) associated genes. These findings suggest that dysregulation of mRNA m6A modification may contribute to the development of AD by affecting the function and gene expression of microglia.

Microglia facilitate and stabilize the response to general anesthesia via modulating the neuronal network in a brain region-specific manner.

General anesthesia leads to a loss of consciousness and an unrousable state in patients. Although general anesthetics are widely used in clinical practice, their underlying mechanisms remain elusive. The potential involvement of nonneuronal cells is unknown. Microglia are important immune cells in the central nervous system (CNS) that play critical roles in CNS function and dysfunction. We unintentionally observed delayed anesthesia induction and early anesthesia emergence in microglia-depleted mice. We found that microglial depletion differentially regulates neuronal activities by suppressing the neuronal network of anesthesia-activated brain regions and activating emergence-activated brain regions. Thus, microglia facilitate and stabilize the anesthesia status. This influence is not mediated by dendritic spine plasticity. Instead, it relies on the activation of microglial P2Y12 and subsequent calcium influx, which facilitates the general anesthesia response. Together, we elucidate the regulatory role of microglia in general anesthesia, extending our knowledge of how nonneuronal cells modulate neuronal activities.

NeuroD1 induces microglial apoptosis and cannot induce microglia-to-neuron cross-lineage reprogramming.

The regenerative capacity of neurons is limited in the central nervous system (CNS), with irreversible neuronal loss upon insult. In contrast, microglia exhibit extraordinary capacity for repopulation. Matsuda et al. (2019) recently reported NeuroD1-induced microglia-to-neuron conversion, aiming to provide an "unlimited" source to regenerate neurons. However, the extent to which NeuroD1 can exert cross-lineage reprogramming of microglia (myeloid lineage) to neurons (neuroectodermal lineage) is unclear. In this study, we unexpectedly found that NeuroD1 cannot convert microglia to neurons in mice. Instead, NeuroD1 expression induces microglial cell death. Moreover, lineage tracing reveals non-specific leakage of similar lentiviruses as previously used for microglia-to-neuron conversion, which confounds the microglia-to-neuron observation. In summary, we demonstrated that NeuroD1 cannot induce microglia-to-neuron cross-lineage reprogramming. We here propose rigid principles for verifying glia-to-neuron conversion. This Matters Arising paper is in response to Matsuda et al. (2019), published in Neuron.

A rapid and quantitative fluorescent microsphere immunochromatographic strip test for detection of antibodies to porcine reproductive and respiratory syndrome virus.

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

Isoliquiritigenin attenuates spinal tuberculosis through inhibiting immune response in a New Zealand white rabbit model.

Spinal tuberculosis (ST) is the tuberculosis caused by Mycobacterium tuberculosis (Mtb) infections in spinal curds. Isoliquiritigenin 4,2',4'-trihydroxychalcone, ISL) is an anti-inflammatory flavonoid derived from licorice (Glycyrrhiza uralensis), a Chinese traditional medicine. In this study, we evaluated the potential of ISL in treating ST in New Zealand white rabbit models. In the model, rabbits (n=40) were infected with Mtb strain H37Rv or not in their 6th lumbar vertebral bodies. Since the day of infection, rabbits were treated with 20 mg/kg and 100 mg/kg of ISL respectively. After 10 weeks of treatments, the adjacent vertebral bone tissues of rabbits were analyzed through Hematoxylin-Eosin staining. The relative expression of Monocyte chemoattractant protein-1 (MCP-1/CCL2), transcription factor κB (NF-κB) p65 in lymphocytes were verified through reverse transcription quantitative real-time PCR (RT-qPCR), western blotting and enzyme-linked immunosorbent assays (ELISA). The serum level of interleukin (IL)-2, IL-4, IL-10 and interferon γ (IFN-γ) were evaluated through ELISA. The effects of ISL on the phosphorylation of IκBα, IKKα/ß and p65 in NF-κB signaling pathways were assessed through western blotting. In the results, ISL has been shown to effectively attenuate the granulation inside adjacent vertebral tissues. The relative level of MCP-1, p65 and IL-4 and IL-10 were retrieved. NF-κB signaling was inhibited, in which the phosphorylation of p65, IκBα and IKKα/ß were suppressed whereas the level of IκBα were elevated. In conclusion, ISL might be an effective drug that inhibited the formation of granulomas through downregulating MCP-1, NF-κB, IL-4 and IL-10 in treating ST.

ATF1 and RAS in exosomes are potential clinical diagnostic markers for cervical cancer.

Cervical cancer is one of the most common cancers among women worldwide. It is highly lethal yet can be treated when found in early stage. Thus, early detection is of significant important for early diagnosis of cervical cancer. Exosomes have been used as biomarkers in clinical diagnosis. It is unknown that whether blood exosomes associated with cervical cancer can be detected and if these exosomes can accurately represent the developmental stage of cervical cancer. Mouse models were made out of a relapsed cervical cancer patient's tumour sample for original and recurrent cervical cancer, and gene analysis in both tumours and exosomes in these mouse models were performed. We found that activating transcription factor 1 (ATF1) and RAS genes were significantly up-regulated in tumours of both primary and recurrent cervical cancer mouse model, and they can also be detected in the blood exosomes of the mouse model. Our results indicated that ATF1 and RAS could be potential candidate biomarkers for cervical cancer in early diagnosis. ATF1 and RAS genes were found significantly elevated in tumours of primary and recurrent cervical cancer mouse model, and they were also detected in the blood exosomes. Therefore, ATF1 and RAS could be used as a diagnostic marker for cervical cancer in the future.

Sidewall cerebral aneurysms: effect of an outflow angle-assisted approach on diagnosis.

BACKGROUND: The ability to diagnose sidewall cerebral aneurysms (SCAs) on an angle measurement basis may be useful in clinical practice. A study was undertaken to evaluate the effect of an outflow angle (OA)-assisted approach. METHODS: MR angiography (MRA) images of 438 patients with suspected SCAs and other cerebrovascular diseases were separately evaluated using the subjective approach and the OA approach. The approaches were then exchanged for confirmation of unclear cases. An OA of ≥90° was considered to represent SCA positivity. The accuracy, sensitivity, and specificity of the OA-assisted approach were determined using patient-based, aneurysm-based, and size-based evaluations. RESULTS: Digital subtraction angiography (DSA) detected 301 SCAs in 267 patients and no SCAs in 171. An OA of ≥90° was observed for 271 aneurysms in 244 patients (true positives); the OA approach misinterpreted OA as <90° for 29 aneurysms in 29 patients (false negatives) and missed one aneurysm. The subjective approach detected 309 SCAs in 273 patients. This approach misdiagnosed 10 patients (false positives) and missed two aneurysms in two patients (false negatives). The OA-assisted approach detected 300 SCAs in 267 patients and no SCAs in 171, overlooking one aneurysm. Patient-based evaluation yielded high accuracy, sensitivity, specificity, and positive and negative predictive values for the OA-assisted approach. CONCLUSIONS: The OA-assisted approach for SCA diagnosis effectively reduced the false-positive rate obtained with the subjective approach with high accuracy, sensitivity, and specificity, suggesting that MRA based on this approach can be a reliable alternative to DSA in SCA screening and diagnosis.

Rapid and sensitive detection of Zika virus by reverse transcription loop-mediated isothermal amplification.

BACKGROUND: Zika virus (ZIKV) is an arbovirus that recently emerged and has expanded worldwide, causing a global threat and raising international concerns. Current molecular diagnostics, e.g., real-time PCR and reverse transcription PCR (RT-PCR), are time consuming, expensive, and can only be deployed in a laboratory instead of for field diagnostics. OBJECTIVES: This study aimed to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) platform showing sensitivity, specificity, and more convenience than previous methods, being easily distributed and implemented. METHODS: Specific primers were designed and screened to target the entire ZIKV genome. The analytical sensitivity and specificity of the assay were evaluated and compared with traditional PCR and quantitative real-time PCR. Three different simulated clinical sample quick preparation protocols were evaluated to establish a rapid and straightforward treatment procedure for clinical specimens in open field detection. RESULTS: The RT-LAMP assay for detection of ZIKV demonstrated superior specificity and sensitivity compared to traditional PCR at the optimum reaction temperature. For the ZIKV RNA standard, the limit of detection was 20 copies/test. For the simulated ZIKV clinical samples, the limit of detection was 0.02 pfu/test, which was one order of magnitude higher than RT-PCR and similar to real-time PCR. The detection limit of simulated ZIKV specimens prepared using a protease quick processing method was consistent with that of samples prepared using commercial nucleic acid extraction kits, indicating that our ZIKV detection method could be used in point-of-care testing. CONCLUSIONS: The RT-LAMP assay had excellent sensitivity and specificity for detecting ZIKV and can be deployed together with a rapid specimen processing method, offering the possibility for ZIKV diagnosis outside of the laboratory.

Up-regulation of KIF14 is a predictor of poor survival and a novel prognostic biomarker of chemoresistance to paclitaxel treatment in cervical cancer.

Kinesin family member 14 (KIF14) is a member of kinesin family proteins which have been found to be dysregulated in various cancer types. However, the expression of KIF14 and its potential prognostic significance have not been investigated in cervical cancer. Real-time PCR was performed to assess the expression levels of KIF14 in 47 pairs of cervical cancer tissues and their matched normal tissues from patients who had not been exposed to chemotherapy as well as tissue samples from 57 cervical cancer patients who are sensitive to paclitaxel treatment and 53 patients who are resistant. The association between KIF14 expression levels in tissue and clinicopathological features or chemosensitivity was examined. Kaplan-Meier analysis and Cox proportional hazards model were applied to assess the correlation between KIF14 expression levels and overall survival (OS) of cervical cancer patients. KIF14 expression levels were significantly increased in cervical cancer tissues compared with matched non-cancerous tissues and it was higher in tissues of patients who are chemoresistant compared with those who are chemosensitive. KIF14 expression was positively associated with high tumour stage (P=0.0044), lymph node metastasis (P=0.0034) and chemoresistance (P<0.0001). Kaplan-Meier analysis showed that high KIF14 expression levels predicted poor survival in patients with (P=0.0024) or without (P=0.0028) paclitaxel treatment. Multivariate analysis revealed that KIF14 was an independent prognostic factor for OS. Our study suggests that KIF14 may serve as a predictor of poor survival and a novel prognostic biomarker of chemoresistance to paclitaxel treatment in cervical cancer.

[Effects of electroacupuncture intervention on changes of quality of ovum and pregnancy out- come in patients with polycystic ovarian syndrome].

OBJECTIVE: To observe the effect of electroacupuncture (EA) treatment on the quality of ovum, stem cell factor(SCF) and the pregnancy outcome in patients with polycystic ovarian syndrome (PCOS), so as to explore its mechanism underlying improving pregnancy rate. METHODS: A total of 200 PCOS patients undergoing in vitro fertilization-embryo transplantation (IVF-ET) were randomly divided into control (medication) group (n = 98) and EA group (n = 102). For patients of the EA group who were undergoing controlled ovarian hyperstimulation, EA stimulation (5 Hz/20 Hz, 15-20 V) was applied to bilateral Shenshu (BL 23), Qihai (CV6), bilateral Zusanli (ST 36)-Sanyinjiao (SP 6), and bilateral Neiguan (PC6)-Zigong (EX-CA 1) for 30 min, once daily till accepting embryo transplant. Patients of the medication group were treated with controlled ovarian hyperstimulation using Diane-35, Decepepty, Gonadotrophin, human Chorionic Gonadotrophin, etc. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2, and progesterone (P) contents were detected using chemiluminescent method. SCF contents in the serum and follicular fluid were assayed by ELISA. The number of retrieved oocytes, fertilization rate, cleavage rate, high quality embryo rate, ovarian hyperstimulation syndrome (OHSS) incidence rate, cycle cancellation rate, clinical pregnancy rate, early abortion rate, Gn dosage and administration duration, and the correlation between the high quality embryo rate and SOF level were determined. RESULTS: Comparison between the two groups showed that the high quality embryo rate, and serum and follicular fluid SCF contents were significantly higher in the EA group than in the medication group (P < 0.05, P < 0.01), while the dosage and administration duration of Gn were significantly lower in the EA group than in the medication group (P < 0.05). A positive correlation was found between the high quality embryo rate and the SCF level in both follicular fluid and serum (P < 0.01). No significant differences were found between the two groups in the number of retrieved oocytes, fertilization rate, cleavage rate, clinical pregnancy rate, OHSS incidence rate, cycle cancellation rate, early abortion rate, serum LH, E2 and P contents (P > 0.05). CONCLUSION: EA can improve the high quality embryo rate, which may be related to its effect in increasing serum and follicular fluid SCF levels.