Proteomics and functional study reveal kallikrein-6 enhances communicating hydrocephalus.

BACKGROUND: Communicating hydrocephalus (CH) is a common neurological disorder caused by a blockage of cerebrospinal fluid. In this study, we aimed to explore the potential molecular mechanism underlying CH development. METHODS: Quantitative proteomic analysis was performed to screen the differentially expressed proteins (DEPs) between patients with and without CH. A CH rat model was verified by Hoechst staining, and the co-localization of the target protein and neuron was detected using immunofluorescence staining. Loss-of-function experiments were performed to examine the effect of KLK6 on the synapse structure. RESULTS: A total of 11 DEPs were identified, and kallikrein 6 (KLK6) expression was found to be significantly upregulated in patients with CH compared with that in patients without CH. The CH rat model was successfully constructed, and KLK6 was found to be co-localized with neuronal nuclei in brain tissue. The expression level of IL-1ß, TNF-α, and KLK6 in the CH group was higher than that in the control group. After knockdown of KLK6 expression using small-interfering RNA (siRNA), the expression levels of synapsin-1 and PSD95 in neuronal cells were increased, and the length, number, and structure of synapses were significantly improved. Following siRNA interference KLK6 expression, 5681 differentially expressed genes (DEGs) were identified in transcriptome profile. The upregulated DEGs of Appl2, Nav2, and Nrn1 may be involved in the recovery of synaptic structures after the interference of KLK6 expression. CONCLUSIONS: Collectively, KLK6 participates in the development of CH and might provide a new target for CH treatment.

Immersion immunization of common carp with bacterial ghost-based DNA vaccine inducing prophylactic protective immunity against spring viraemia of carp virus.

The interactive applications of immunization route, vaccine type and delivery vectors are emerging as a key area of research within the field of mass immunization in fishery production. In an effort to improve DNA vaccine's immune efficiency in large-scale immunization, a promising bacterial ghost-loaded DNA vaccine was constructed based on Escherichia coli DH5α. In common carp was investigated the immune response to immersion immunization via related indicator analysis, and the challenge test of spring viraemia of carp virus (SVCV) was carried out. The result indicated that BG-loaded DNA vaccine induced higher serum antibody level than naked pEG-G. Simultaneously, the immunophysiological indicators and genes change at the more advanced levels in the BG/pEG-G immune group. At the treatment concentration of 20 mg/L of the BG/pEG-G group, IgM and IgZ expressions in vivo were markedly increased by 21.62 times and 6.91 times, respectively, and the relative percentage survival reached the peak of 59.57%. This study paves the way for future aquatic animal vaccine research, which aimed to develop the highly effective immersion vaccine system by delivery vectors, with the ultimate aim to prevent and restrict SVCV in actual production.

Neuronal Apoptosis Preventive Potential of Sophocarpine via Suppression of Aß-Accumulation and Down-Regulation of Inflammatory Response.

In the current study sophocarpine was investigated in vitro for prevention of ß-amyloid induced PC12 neuronal cell damage. Exposure to ß-amyloid caused a dose-dependent suppression in growth of PC12 cells with maximum reduction at 10 µM. Sophocarpine pre-treatment reversed suppressive effect of ß-amyloid (10 µM) on PC12 cell growth in concentration-based manner. In sophocarpine pre-treated PC12 cells the ß-amyloid mediated PGE2 level elevation was attenuated significantly at 0.25-2 µM doses. Moreover, in sophocarpine pretreated PC12 cells the ß-amyloid mediated promotion of COX-2 level was also inhibited. Sophocarpine pre-treatment attenuated iNOS expression in ß-amyloid exposed PC12 cells at 0.25-2 µM doses. Pre-treatment of PC12 cells with sophocarpine suppressed NO-species generation induced by ß-amyloid exposure. In sophocarpine pretreated PC12 cells elevation of nuclear NF-κB expression induced by ß-amyloid was significantly inhibited. In summary, sophocarpine prevents reduction of PC12 cell growth induced by ß-amyloid exposure via inhibition of inflammatory processes. The preventive effect of sophocarpine on ß-amyloid induced PC12 cell damage is associated with inhibition of NF-κB nuclear translocation. Therefore, sophocarpine may be used for treatment of neurological disorders like Alzheimer's disease.

Distinct roles of neuregulin in different models of neuropathic pain.

The objective of the study was to investigate the role of neuregulin-ErbB signaling in neuropathic pain in different types of injury. Neuregulin-1(NRG-1) was injected into animals with either formalin-induced pain model or spared nerve injury (SNI) model. Formalin tests or paw withdrawal tests were performed to study the role of NRG-1 in neuropathic pain. siRNA specific to different erbB receptors were then introduced to test which specific signaling pathway was required for NRG-1 signaling in the different pain models. NRG-1 inhibits neuropathic pain after SNI in a dose-dependent manner, while NRG-1 aggravates formalin-induced neuropathic pain. ErbB2 and erbB4 receptors were activated after neuregulin administration. Knockdown of ErbB2 relieves the aggravation of NRG-1 on formalin-induced neuropathic pain, and knockdown of ErbB4 could relieve the inhibition of NRG-1 on neuropathic pain in the SNI model. NRG-1 has two distinct functions depending on the different receptor activation in different models of neuropathic pain. These novel findings may provide new therapeutic approaches for the treatment of neuropathic pain in different injury types.

Impact of Endoscopic Surgery Versus Robot CAS-R-2 Assisted with Stereotactic Drainage on Prognosis of Basal Ganglia Hypertensive Intracerebral Hemorrhage.

OBJECTIVE: To evaluate the impact of endoscopic surgery (ES) versus robot CAS-R-2 assisted with stereotactic drainage on prognosis of basal ganglia hypertensive intracerebral hemorrhage (HICH). METHODS: This retrospective observational study included patients who underwent ES or robot CAS-R-2 assisted with stereotactic drainage for basal ganglia HICH in Shanghai Sixth People's Hospital between June 2017 and May 2022. The outcomes were 6-month mortality and modified Rankin Scale (mRS) score. RESULTS: A total of 94 patients were included; 68 (age 51.26 ± 9.18 years, 17 women) of them underwent ES, while the other 26 (age 56.50 ± 12.91, 11 women) underwent robot CAS-R-2. The 6-month mortality rates were similar (P > 0.05) between the patients who underwent ES (6 of 68, 8.82%) and robot CAS-R-2 (2 of 26,7.69%), while the rate of good prognosis in the ES group was significantly higher compared with that in the robot CAS-R-2 group (P = 0.024). Univariate logistic analysis found that endoscopic surgery, age, and hematoma volume were associated with poor prognosis at 6 months. Multivariate logistic regression analysis showed that, after adjusted for the preoperative hematoma volume and age, endoscopy surgery (relative risk 0.21, 95% CI 0.06-0.68, P = 0.009) was associated with good prognosis at 6 months follow-up. CONCLUSIONS: Compared with robot CAS-R-2 assisted with stereotactic drainage, ES might have higher rate of good prognosis at 6-month follow-up for basal ganglia HICH.

Research progress in the preparation, structural characterization, and biological activities of polysaccharides from traditional Chinese medicine.

Traditional Chinese medicines are tremendous sources of polysaccharides, which are of great interest in the human welfare system as natural medicines, food, and cosmetics. This review aims to highlight the recent trends in extraction (conventional and non-conventional), purification and analytic techniques of traditional Chinese medicine polysaccharides (TCMPs), and the chemical structure, biological activities (anti-tumor, hypoglycemic, antioxidant, intestinal flora regulation, immunomodulatory, anti-inflammatory, anti-aging, hypolipidemic, hepatoprotective, and other activities), and the underlying mechanisms of polysaccharides extracted from 76 diverse traditional Chinese medicines were compared and discussed. With this wide coverage, a total of 164 scientific articles were searched from the database including Google Scholar, PubMed, Web of Science, and China Knowledge Network. This comprehensive survey from previous reports indicates that TCMPs are non-toxic, highly biocompatible, and good biodegradability. Besides, this review highlights that TCMPs may be excellent functional factors and effective therapeutic drugs. Finally, the current problems and future research advances of TCMPs are also introduced. New valuable insights for the future researches regarding TCMPs are also proposed in the fields of therapeutic agents and functional foods.

Rationally Designed Self-Assembling Nanovaccines Elicit Robust Mucosal and Systemic Immunity against Rhabdovirus.

Viral diseases have constantly caused great threats to global public health, resulting in an urgent need for effective vaccines. However, the current viral vaccines often show low immunogenicity. To counter this, we report a smart strategy of a well-designed modular nanoparticle (LSG-TDH) that recapitulates the dominant antigen SG, low-molecular-weight protamine, and tetralysine-modified H-chain apoferritin (TDH). The constructed LSG-TDH nanovaccine could self-assemble into a nanocage structure, which confers excellent mucus-penetrating, cellular affinity, and uptake ability. Studies demonstrate that the LSG-TDH nanovaccine could strongly activate both mucosal and systemic immune responses. Importantly, by immunizing wild-type and TLR2 knockout (TLR2-KO) zebrafish, we found that TLR2 could mediate LSG-TDH-induced adaptive mucosal and systemic immune responses by activating antigen-presenting cells. Collectively, our findings offer new insights into rational viral vaccine design and provide additional evidence of the vital role of TLR2 in regulating adaptive immunity.

Research on the correlation between three-dimensional morphology and temperature changes in potato slices during drying.

Drying is an effective method to reduce potato storage loss. However, potatoes have high porosity with high water content. Shrinkage during drying can lead to folding and cracking of the dried product form. Therefore, this study explored the correlation between the 3D morphology and temperature distribution changes of potato slices during drying, with the aim of providing a reference for the detection of quality changes. An online automatic acquisition device to obtain 3D morphology and temperature information was designed and built. Hot air-drying experiments were conducted on the potato slices. 3D morphology images and temperature images of the potato slices were acquired by 3D and temperature sensors, and the two images were registered using the random sample consensus (RANSAC) algorithm. The region of interest of each image was extracted by algorithms such as threshold segmentation, hole filling and morphological erosion, and the 3D morphology information and temperature information were obtained. The mapping, range and average of each acquisition point were calculated for correlation analysis. Spearman's rank correlation coefficients and Maximum Information Coefficient (MIC) values were selected as measures for the correlation study. The results showed that the Spearman's rank correlation coefficients between average height and average temperature were mostly above 0.7 in absolute value, and the MICs were mostly above 0.9. The average values of the 3D information and temperature information exhibited an extremely strong correlation. This paper gives a new approach to investigate the morphological changes in the drying process by quantifying the relationship between 3D morphology and temperature distribution. This can guide the improvement of potato drying and processing methods.

Jiedu Yizhi Formula Improves Cognitive Function by Regulating the Gut Dysbiosis and TLR4/NF-κB Signaling Pathway.

Objective: The objective of this study was to explore the neuroprotective mechanism of JDYZF in treating AD from the perspective of inflammation and intestinal microflora. Methods: A total of 24 APP/PS1 mice were randomly divided into four groups: model (n = 6), JDYZF low-dose (n = 6), JDYZF high-dose (n = 6), and positive drug (n = 6), six C57 mice were used as the control group. The body weights and diets of all mice were examined daily. After 8 weeks of administration, the learning and memory of mice were evaluated by the Morris water maze test. The histopathological changes of hippocampus, liver and kidney in mice were observed by HE staining after being euthanized. The expression of p-tau in hippocampus tissue was detected by immunohistochemistry. After that, 16S rDNA sequencing was used to investigate the relationship between JDYZF and intestinal microbiota. Finally, a comparison of TLR4, p65, p-p65, iκB, p-iκB, and IL-1ß protein expression in the hippocampus tissue of mice in each group was measured by Western blot. Results: The results showed that APP/PS1 mice taking JDYZF orally were generally in good condition. Compared with the control group, JDYZF significantly improved learning and memory ability in ethology. Histology showed that JDYZF improved the hippocampal structure of mice and inhibited the deposition of p-tau. JDYZF treatment could regulate the gut microbiota of APP/PS1 mice by increasing the richness of Lachnospiraceae, Ruminococcaceae, and Actinobacteria and reducing that of Alistipes and Muribaculaceae. It also significantly inhibited the activation of the TLR4/NF-κB signaling pathway in the brain. In addition, no obvious toxic reactions were found in the liver and kidney of APP/PS1 mice after taking JDYZF for 8 weeks. Conclusion: The findings revealed that JDYZF improved cognitive ability and alleviated the TLR4/NF-κB signaling pathway in APP/PS1 mice, and the modulating the gut microbiota presented here may help illuminate its activation mechanism.

Study on neutron energy spectrum unfolding algorithm with EJ309 liquid scintillation detector.

Proton recoil method can be used to experimentally measure fast neutron energy spectrum of non-pulsed neutron sources. The neutron energy spectrum unfolding algorithms based on the MLEM method, the GOLD deconvolution method, the Direct-D method, have been developed by using the EJ309 liquid scintillation detector. The degree of iteration by the mean square error (MSE) is proposed as a judgment criterion by according to the iterative accuracy, convergence speed and iteration efficiency. The developed neutron energy spectrum unfolding algorithms can unfolding the standard simulated mono-energetic neutron spectrum (2.5 MeV), 252Cf neutron spectrum, Am-Be neutron spectrum and the experimentally measured D-D neutron spectrum with higher precision as well as fewer iterations. The unfolded neutron spectra are in good agreement with the standard simulated neutron spectra and evaluated D-D neutron spectrum, which is revealed that the developed unfolding algorithms can unfolding neutron energy spectrum with reasonable accuracy.

miR-210 activates notch signaling pathway in angiogenesis induced by cerebral ischemia.

The compensatory angiogenesis that occurs after cerebral ischemia increases blood flow to the injured area and limits extension of the ischemic penumbra. In this way, it improves the local blood supply. Fostering compensatory angiogenesis is an effective treatment for ischemic cerebrovascular disease. However, angiogenesis in the adult organism is a complex, multi-step process, and the mechanisms underlying the regulation of angiogenesis are not well understood. Although Notch signaling reportedly regulates the vascularization process that occurs in ischemic tissues, little is known about the role of Notch signaling in the regulation of ischemia-induced angiogenesis after ischemic stroke. Recent research has indicated that miR-210, a hypoxia-induced microRNA, plays a crucial role in regulating the biological processes that occur in blood vessel endothelial cells under hypoxic conditions. This study was undertaken to investigate the role of miR-210 in regulating angiogenesis in response to brain ischemia injury and the role of the Notch pathway in the body's response. We found miR-210 to be significantly up-regulated in adult rat ischemic brain cortexes in which the expression of Notch1 signaling molecules was also increased. Hypoxic models of human umbilical vein endothelial cells (HUVE-12) were used to assess changes in miR-210 and Notch1 expression in endothelial cells. Results were consistent with in vivo findings. To determine the molecular mechanisms behind these phenomena, we transfected HUVE-12 cells with miR-210 recombinant lentiviral vectors. We found that miR-210 overexpression caused up-regulation of Notch1 signaling molecules and induced endothelial cells to migrate and form capillary-like structures on Matrigel. These data suggest that miR-210 is involved in the regulation of angiogenesis in response to ischemic injury to the brain. Up-regulation of miR-210 can activate the Notch signaling pathway, which may contribute to angiogenesis after cerebral ischemia.

A prospective clinical study of routine repeat computed tomography (CT) after traumatic brain injury (TBI).

PURPOSE: To discuss the repeated CT scanning in patients with traumatic brain injury (TBI) and to identify the conditions under which this approach is necessary. METHODS: One hundred and seventy-one patients who suffered TBI but were not surgically treated were divided into two groups: the routine-repeat CT group (n = 89) and the non-routine-repeat CT group (n = 82). The patients' clinical characteristics were compared. T-tests and stepwise logistic regression were used for analysis. Patients in the routine-repeat CT group were divided into three groups according to GCS scores to determine the need for routinely repeated CT scans. RESULTS: The results revealed statistically significant differences between the two groups in terms of neuro-ICU-LOS and LOS (p < 0.01). No significant differences emerged with respect to hospital charges and GCS scores at discharge (p > 0.05). AGE, international normalized ratio (INR), D-dimer concentration (DD), GCS scores and number of hours between the first CT scan and the injury (HCT1) were influential factors of developing progressive haemorrhage. CONCLUSION: The routine-repeat CT group fared better than did the non-routine-repeat CT group. Routinely repeated CTs were minimally effective among those with mild TBI, whereas this procedure demonstrated a significant effect on patients with moderate and severe TBI.

LINC00662 Promotes Proliferation and Invasion and Inhibits Apoptosis of Glioma Cells Through miR-483-3p/SOX3 Axis.

LINC00662 plays a prominent role in the carcinogenesis and progression of diverse cancers. However, its biological functions in glioma are still unclear. LINC00662 expression in glioma tissue samples and cell lines was examined by quantitative real-time polymerase chain reaction. The correlation between LINC00662 expression and the clinical characteristics of 50 patients with glioma was analyzed. LINC00662 knockdown and overexpression cell lines were constructed, and the effects of LINC00662 on the proliferation, invasion, and apoptosis of glioma cells were evaluated by cell counting kit-8, 5-ethynyl-2'-deoxyuridine, Transwell, and flow cytometry assays, respectively. Besides, the relationships among LINC00662, miR-483-3p, and sex-determining region Y-box 3 (SOX3) were assessed by dual-luciferase reporter assay and RNA immunoprecipitation assay. Western blot was used to detect the regulatory effects of LINC00662 and miR-483-3p on SOX3 expression in glioma cells. LINC00662 expression level was elevated in glioma tissues and cell lines compared to that in normal tissues and cell lines. LINC00662 high expression was associated with the adverse prognosis of patients with glioma. Knockdown of LINC00662 repressed the proliferation and invasion of glioma cells, and promoted apoptosis. Additionally, it was revealed that LINC00662 acted as the molecular sponge of miR-483-3p, and SOX3 was verified as a direct target of miR-483-3p. The inhibition of miR-483-3p expression and SOX3 overexpression reversed the biological effects of LINC00662 knockdown on glioma cells. This study reports the key regulatory role of LINC00662/miR-483-3p/SOX3 axis in the tumorigenesis and progression of glioma, bringing novel insights into the underlying mechanisms of glioma.

TLR2-mediated mucosal immune priming boosts anti-rhabdoviral immunity in early vertebrates.

Most pathogens utilize mucosal surfaces to enter and propagate within the host. As one of the main pathogens of fatal and highly contagious diseases, rhabdoviruses are distributed widely in nature affecting both human and animals. Therefore, local mucosal immune responses, most effectively induced by mucosal vaccines, act as frontline immunity to block the pathogens at its initial replication sites. However, the underlying regulatory mechanisms of mucosal immunity triggered by mucosal vaccine remains unclear. Herein, a rhabdoviruses glycoprotein-based mucosal vaccine (G131c) was used to elucidate the regulatory mechanism of local mucosal immunity in zebrafish, a typical immunological model. Firstly, we verified the strong immunoprotection of G131c mucosal vaccine. Furthermore, the delivery kinetics of G131c was evaluated in vivo, indicating the effective uptake of vaccines by mucosal tissues through immersion vaccination. Importantly, we demonstrate immersion with G131c vaccine could activate antigen presenting cells (APCs) at the local mucosal sites, and then arose robust local mucosal and systemic immune responses. More critically, we found that G131c mediated these immune effects by interacting with Toll-like receptor 2 (TLR2) and activating downstream MAPK and NF-κB signaling pathways. Thus, our findings provide previously unappreciated evidence that rhabdovirus glycoprotein could interact with TLR2 and then activate the APCs in local mucosal sites. This study provides a comprehensive perspective on the mechanism of TLR2-mediated mucosal immunity in the early vertebrates.

Jiedu-Yizhi Formula Improves Cognitive Impairment in an Aß 25-35-Induced Rat Model of Alzheimer's Disease by Inhibiting Pyroptosis.

Jiedu-Yizhi formula (JDYZF) is prescribed for the treatment of Alzheimer's disease (AD) and was created by Jixue Ren, a master of traditional Chinese medicine, based on the "marrow deficiency and toxin damage" theory. In our clinic, this formula has been used for the treatment of AD for many years and has achieved good results. However, the mechanism by which JDYZF improves cognitive impairment has not been determined. In this study, we confirmed that orally administered JDYZF reversed the cognitive deficits in an Aß 25-35-induced rat model, increased the number of neurons in the hippocampal CA1 area, improved their structure, decreased the deposition of ß-amyloid (Aß), reduced the expression of proteins related to the NLRP3/Caspase-1/GSDMD and LPS/Caspase-11/GSDMD pyroptosis pathways, and reduced the levels of interleukin 1ß (IL-1ß) and IL-18, thereby inhibiting the inflammatory response. In addition, JDYZF exerted no hepatotoxicity in rats. In short, these results provide scientific support for the clinical use of JDYZF to improve the cognitive function of patients with AD.

Dual-Targeting Polymer Nanoparticles Efficiently Deliver DNA Vaccine and Induce Robust Prophylactic Immunity against Spring Viremia of Carp Virus Infection.

Spring viremia of carp virus (SVCV) is highly contagious and lethal to most cyprinid fish, causing serious economic losses to the carp aquaculture industry. Although DNA vaccines can generate long-term humoral and cellular immune responses, which provide protective immunity against SVCV, the major drawback of DNA vaccines is their low immunogenicity in clinical tests. Here, we construct a dual-targeted polymer DNA vaccine delivery platform (MCS-PCHG) by using mannosylated chitosan to encapsulate the poly(d,l-lactide-co-glycolide)-loaded DNA vaccine containing the heavy-chain CH3 region (CH3) of common carp IgM and the antigenic domain (G131c). The developed nanovaccine delivery platform showed good biocompatibility in vivo and in vitro. With the modification of the mannose moiety and the modification of CH3, the constructed MCS-PCHG could efficiently activate the maturation of antigen-presenting cells. Moreover, we observe significantly high level of immune-related genes expression, serum antigen-specific IgM, SVCV-neutralizing antibody titers in fish vaccinated with MCS-PCHG. Next, the protective efficacy of MCS-PCHG was further evaluated by challenge test. The highest survival rate (ca. 84%) was observed in fish vaccinated with MCS-PCHG after challenging with SVCV. This study presents a novel design for smart, dual-targeted polymer nanoparticles, which are inherently biocompatible, promising for targeted vaccine delivery. IMPORTANCE Spring viremia of carp virus (SVCV) affects global cyprinid fish farming industry, with no available commercial vaccine. Herein, we developed a dual-targeting polymer nanovaccine (MCS-PCHG) by using mannose and common carp IgM heavy chain CH3 region (CH3) as antigen presenting cell (APCs) recognition moiety, attaining the effective delivery of antigen. This dual-targeting polymer vaccine can efficiently activate the APCs, and further induce robust and durable adaptive immune response with good protection against SVCV infection. Our study provides valuable theoretical basis for developing efficient vaccine against infectious diseases in aquaculture.

Eggshell crack detection based on the transient impact analysis and cross-correlation method.

To explore the transient impact process for cracked eggshell detection, an equivalent mechanical model was built based on a self-designed automatic excitation device. Through analysis of power spectrum from dynamic force signal, it was found that the impact speed affects only the impaction energy. In contrast, the material of the impact head and the weight of the excitation rod determine the energy and the cut-off frequency of the impaction. When the weight of impact tup is less than 6.62 g, the cut-off frequency of excitation impact can cover the egg's inherent frequency. Then, an optimized experiment system was designed to acquire the response acoustic signals. The cross-correlation analysis and Bayes classification methods were carried out to detect the cracked eggshell. In the conducted experiments, a crack detection level of 97% and a false rejection level of 1% were achieved. From the findings, it can be concluded that the proposed method will assist in optimizing the impact device and simplifying the classification algorithm for an online detection system.

Three-dimensional morphological changes of potato slices during the drying process.

Hot air drying is a common method for drying potato slices. In this paper, the three-dimensional (3D) morphological changes incurred during hot air drying of potato slices were investigated. The effect of drying on the thickness and diameter of potato slices was of special interest. The results showed that the potato slices underwent stages of regular warping, collapse, and curling during the drying process. After classifying the numerical variation in characteristics into the standard deviation in mean height (SDMH) and the rate of change in mean height (RCMH) of potato slices, the RCMH was selected to describe the 3D morphological changes in the potato drying process. A critical point and a termination point for RCMH of potato slices in the drying process were observed. Samples varied widely after the critical point was reached. A logarithmic function model was used to assess differences in the RCMH at the critical point and the termination point. The R-squared (R2) value of 0.9 suggested a strong correlation between the parameters of the experiment and changes in slice thickness and diameter. The model proposed in this paper could accurately characterize the late-stage changes in potato slice qualities during hot air drying of potato slices.

miR-29a-5p Alleviates Traumatic Brain Injury- (TBI-) Induced Permeability Disruption via Regulating NLRP3 Pathway.

OBJECTIVE: Inactivation of NLRP3 inflammasome plays a role in reducing the permeability of endothelial cells and improving blood-brain barrier (BBB) dysfunction following traumatic brain injury (TBI). However, the mechanism controlling NLRP3 inflammasome activation remains unclear. This study is aimed at defining the role of miR-29a-5p in NLRP3 inflammasome activation and permeability of endothelial cells under TBI. METHODS: The scratch injury model on brain bEnd.3 microvascular endothelial cells was used as in vitro TBI model cells. Effects of miR-29a mimics and inhibitors on TBI model cells were observed by examining their action on FITC, TEER, and protein contents of ZO-1 and occludin, and cell permeability-associated protein. Luciferase reporter assay evaluated miR-29a-5p targeting to NLRP3. ELISA examined of IL-1ß and IL-18 levels. miR-29a-5p mimic was injected into TBI mouse and its effect on BBB, indicated by Evans blue (EB) staining assay and cerebral water content, and NLRP3 activation was examined. RESULTS: miR-29a-3p and miR-29a-5p mimics decrease the concentration of FITC, and increase TEER and the protein contents of ZO-1 and occludin in TBI model cells. miR-29a-5p silencing disrupted the permeability of mouse bEnd.3 cells. miR-29a-5p targets to NLRP3 through the binding on its 3'UTR and negatively regulates its expression in TBI model cells. NLRP3 inhibition and miR-29a-5p silencing together caused significantly decreased FITC concentration and increased TEER value and release of IL-1ß and IL-18. miR-29a-5p mimic alleviated the BBB and cerebral water content and inactivates NLRP3 in the mouse TBI model. CONCLUSIONS: miR-29a-5p mimics protect TBI-induced increased endothelial cell permeability and BBB dysfunction via suppressing NLRP3 expression and activation.

Proteomic Analysis Reveals that Di Dang Decoction Protects Against Acute Intracerebral Hemorrhage Stroke in Rats by Regulating S100a8, S100a9 Col1a1, and Col1a2.

OBJECTIVE: The present study aimed to explore the neuroprotective mechanism of Di Dang decoction (DDD) during acute intracerebral hemorrhage (AICH) stroke in Sprague Dawley rats through proteomic analysis. METHODS: A total of 135 healthy Sprague Dawley rats were randomly divided into five groups: control (n = 27), model (n = 27), DDD low-dose (n = 27), DDD medium-dose (n = 27), and DDD high-dose (n = 27). AICH stroke in rats was induced by injecting autologous blood into the caudate nucleus. The modified Neurological Severity Score (mNSS) was used to evaluate the cerebral nerve function deficit. Hematoxylin and eosin (HE) staining was performed to observe the brain tissue at the lesion site. Albumin concentration was assessed on obvious blood-brain barrier damaged and brain water content was used to evaluate the brain injury. For quantitative proteomics, proteins were extracted from the cerebral cortices. Target proteins were identified using mass spectrometer-based targeted proteomic quantification. RESULTS: mNSS score, HE staining results, albumin concentration, and brain water content showed the most significant improvements in the neuroprotective in the high-dose group 7 days after DDD exposure. Furthermore, quantitative proteomics analysis showed that, relative to the control group, S100a8 and S100a9 were downregulated by 0.614 (p = 0.033702) and 0.506 times (p = 0.000024) in the high-dose group. Compared with the control group, Col1a1 and Col1a2 were upregulated by 1.319 (p = 0.000184) and 1.348 (p = 0.014097) times in the high-dose group. These results were confirmed using mass spectrometer-based targeted proteomic quantification. CONCLUSION: Application of a high-dose DDD for 7 days in AICH stroke rats showed the most significant improvements in neuroprotective. Mechanistically, this effect was mediated by S100a8 and S100a9 protein downregulation and Col1a1 and Col1a2 upregulation.