Dl-3-n-Butylphthalide Alleviates Secondary Brain Damage and Improves Working Memory After Stroke in Cynomolgus Monkeys.

BACKGROUND: Remote secondary neurodegeneration is associated with poststroke cognitive impairment (PSCI). Dl-3-n-butylphthalide (NBP) improves PSCI clinically. However, whether it ameliorates PSCI by alleviating secondary neurodegeneration remains uncertain. Nonhuman primates provide more relevant models than rodents for human stroke and PSCI. This study investigated the effects of NBP on PSCI and secondary neurodegeneration in cynomolgus monkeys after permanent left middle cerebral artery occlusion (MCAO). METHODS: Thirteen adult male cynomolgus monkeys were randomly assigned to sham (n=4), MCAO+placebo (n=5), and MCAO+NBP groups (n=4). The MCAO+placebo and MCAO+NBP groups received saline and NBP injections intravenously, respectively, starting at 6-hour postsurgery for 2 weeks, followed by soybean oil and NBP orally, respectively, for 10 weeks after MCAO. Infarct size was assessed at week 4 by magnetic resonance imaging. Working memory and executive function were evaluated dynamically using the delayed response task and object retrieval detour task, respectively. Neuron loss, glia proliferation, and neuroinflammation in the ipsilateral dorsal lateral prefrontal cortex, thalamus, and hippocampus were analyzed by immunostaining 12 weeks after MCAO. RESULTS: Infarcts were located in the left middle cerebral artery region, apart from the ipsilateral dorsal lateral prefrontal cortex, thalamus, or hippocampus, with no significant difference between the MCAO+placebo and MCAO+NBP group. Higher success in delayed response task was achieved at weeks 4, 8, and 12 after NBP compared with placebo treatments (P<0.05), but not in the object retrieval detour task (all P>0.05). More neurons and less microglia, astrocytes, CD68-positive microglia, tumor necrosis factor-α, and inducible NO synthase were observed in the ipsilateral dorsal lateral prefrontal cortex and thalamus after 12 weeks of NBP treatment (P<0.05), but not in the hippocampus (P>0.05). CONCLUSIONS: Our findings indicate that NBP improves working memory by alleviating remote secondary neurodegeneration and neuroinflammation in the ipsilateral dorsal lateral prefrontal cortex and thalamus after MCAO in cynomolgus monkeys.

Transcription factor LBD16 targets cell wall modification/ion transport genes in peach lateral root formation.

LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKEs (LBDs/ASLs) are plant-specific transcription factors that function downstream of auxin-regulated lateral root (LR) formation. Our previous research found that PpLBD16 positively regulates peach (Prunus persica) LR formation. However, the downstream regulatory network and target genes of PpLBD16 are still largely unknown. Here, we constructed a PpLBD16 homologous overexpression line and a PpLBD16 silenced line. We found that overexpressing PpLBD16 promoted peach root initiation, while silencing PpLBD16 inhibited peach root formation. Through RNA sequencing (RNA-seq) analysis of roots from PpLBD16 overexpression and silenced lines, we discovered that genes positively regulated by PpLBD16 were closely related to cell wall synthesis and degradation, ion/substance transport, and ion binding and homeostasis. To further detect the binding motifs and potential target genes of PpLBD16, we performed DNA-affinity purification sequencing (DAP-seq) analysis in vitro. PpLBD16 preferentially bound to CCNGAAANNNNGG (MEME-1), [C/T]TTCT[C/T][T/C] (MEME-2), and GCGGCGG (ABR1) motifs. By combined analysis of RNA-seq and DAP-seq data, we screened candidate target genes for PpLBD16. We demonstrated that PpLBD16 bound and activated the cell wall modification-related genes EXPANSIN-B2 (PpEXPB2) and SUBTILISIN-LIKE PROTEASE 1.7 (PpSBT1.7), the ion transport-related gene CYCLIC NUCLEOTIDE-GATED ION CHANNEL 1 (PpCNGC1) and the polyphenol oxidase (PPO)-encoding gene PpPPO, thereby controlling peach root organogenesis and promoting LR formation. Moreover, our results displayed that PpLBD16 and its target genes are involved in peach LR primordia development. Overall, this work reveals the downstream regulatory network and target genes of PpLBD16, providing insights into the molecular network of LBD16-mediated LR development.

Disparate Radiation-Induced Microstructural Injuries in Whole-Brain White Matter of Patients With Nasopharyngeal Carcinoma: A Longitudinal Study Using Multishell Diffusion MRI.

BACKGROUND: Evidence for prevention strategies of radiotherapy (RT)-related injury in patients with nasopharyngeal carcinoma (NPC) was lacking. Understanding the dynamic alterations in the cerebral white matter (WM) microstructure after RT may be helpful. PURPOSE: To investigate the dynamic alterations in the whole brain WM microstructure in patients with NPC in the 12 months after RT using multishell diffusion MRI (MS-dMRI). STUDY TYPE: Single-center longitudinal study. POPULATION: A total of 28 treatment-naïve patients with pathologically confirmed NPC (age: 39.68 ± 8.93 years, 11 female) and 20 healthy controls (age: 40.65 ± 9.76 years, 7 female). FIELD STRENGTH/SEQUENCES: A 3 T, MS-dMRI using a single-shot echo planar imaging sequence. ASSESSMENT: MS-dMRI was acquired at baseline for the NPC patients and healthy controls, at 0-3 (acute, AC), 6 (early delayed, ED) and 12 months (late delayed, LD) after RT for the NPC patients. The mean and maximum radiation doses to the temporal lobe were acquired. The quality of images was reviewed. MS-dMRI was analyzed using tract-based spatial statistics (TBSS). The presentations of injury were defined by the findings of TBSS. STATISTICAL TESTS: Chi-square, t tests, repeated ANOVA, and Spearman-rank correlation analysis were used. P < 0.05 was considered to be statistically significant. RESULTS: TBSS showed two WM injuries (injuries 1 and 2). Injury 1 emerged in the ED phase in the bilateral temporal poles and persisted throughout the ED and LD phases. Injury 2 developed from the AC to ED phase in the bilateral hemisphere and partially recovered in the LD phase. In the ED and LD phases, the multiple diffusion metrics were well correlated (r > 0.5 or <-0.5) with the RT dose, especially in the WM tracts in the temporal lobes. DATA CONCLUSION: Disparate WM injuries were observed in NPC patients after RT. The injuries may be primarily or secondarily induced by radiation. Injury 1 may be irreversible, while injury 2 seems to partially recover. EVIDENCE LEVEL: 2. TECHNICAL EFFICACY: Stage 4.

Health risk assessment of heavy metals based on source analysis and Monte Carlo in the downstream basin of the Zishui.

In this study, stone coal mines in the lower reaches of the Zijiang River were adopted as the research object. To analyze the spatial distribution, sources, and health risks of heavy metals in the surrounding soil of stone coal mines, 82 topsoil samples were collected in the study area, and the contents of 8 heavy metals including Cd, Hg, As, Cr, Pb, Cu, Ni, and Zn were determined. The spatial distribution of heavy metals was analyzed using ArcGIS, and the pollution sources of heavy metals were identified using Positive matrix factorization (PMF). Then, Monte Carlo and health risk assessment models were used to evaluate the health risks of different populations. The results showed that the average content of heavy metals followed the order of Zn > Cr > Pb > Cu > Ni > As > Cd > Hg, and the contents of all heavy metals were higher than the soil background values of Hunan Province. The high-value areas of heavy metals content were mostly concentrated in the central region close to areas with a notable concentration of stone coal mines. PMF identified four pollution sources, namely, mining activities (26.9%), atmospheric deposition (18.8%), natural sources (32.8%) and agricultural sources (21.5%). The carcinogenic and non-carcinogenic risks for children were higher than those for adults, with As and Cd posing higher carcinogenic risks to children. Based on the source of health risks, it was determined that the health risks could be primarily attributed to agricultural sources, and As was the main heavy metal causing health risks. This study provides theoretical support for treating heavy metal pollution in mining basins.

Identification of disulfidptosis-related subtypes, characterization of tumor microenvironment infiltration, and development of a prognosis model in breast cancer.

Introduction: Breast cancer (BC) is now the most common type of cancer in women. Disulfidptosis is a new regulation of cell death (RCD). RCD dysregulation is causally linked to cancer. However, the comprehensive relationship between disulfidptosis and BC remains unknown. This study aimed to explore the predictive value of disulfidptosis-related genes (DRGs) in BC and their relationship with the TME. Methods: This study obtained 11 disulfidptosis genes (DGs) from previous research by Gan et al. RNA sequencing data of BC were downloaded from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus database (GEO) databases. First, we examined the effect of DG gene mutations and copy number changes on the overall survival of breast cancer samples. We then used the expression profile data of 11 DGs and survival data for consensus clustering, and BC patients were divided into two clusters. Survival analysis, gene set variation analysis (GSVA) and ss GSEA were used to compare the differences between them. Subsequently, DRGs were identified between the clusters used to perform Cox regression and least absolute shrinkage and selection operator regression (LASSO) analyses to construct a prognosis model. Finally, the immune cell infiltration pattern, immunotherapy response, and drug sensitivity of the two subtypes were analyzed. CCK-8 and a colony assay obtained by knocking down genes and gene sequencing were used to validate the model. Result: Two DG clusters were identified based on the expression of 11DGs. Then, 225 DRGs were identified between them. RS, composed of six genes, showed a significant relationship with survival, immune cell infiltration, clinical characteristics, immune checkpoints, immunotherapy response, and drug sensitivity. Low-RS shows a better prognosis and higher immunotherapy response than high-RS. A nomogram with perfect stability constructed using signature and clinical characteristics can predict the survival of each patient. CCK-8 and colony assay obtained by knocking down genes have demonstrated that the knockdown of high-risk genes in the RS model significantly inhibited cell proliferation. Discussion: This study elucidates the potential relationship between disulfidptosis-related genes and breast cancer and provides new guidance for treating breast cancer.

Contralateral internal mammary artery perforator flap for large inner chest wall defect repair and immediate breast reconstruction after excision of large malignant inner breast tumors.

OBJECTIVES: Large breast tumor resection can cause chest wall defects that are difficult to close. A combination of oncoplastic techniques is required to repair chest wall defects and immediately reconstruct the breast. In this report, we present the use of the contralateral internal mammary artery perforator (IMAP) flap to repair large chest wall defects and perform breast reconstruction for a series of patients. METHODS: Between July 2013 and June 2020, 10 patients and 4 patients underwent chest wall defect repair and breast reconstruction, respectively, with contralateral IMAP flaps. The average body mass index was 26.9 kg/m2 (range: 21.5-33.7 kg/m2 ). Tumor sizes varied from 2.5 × 1.5 × 0.8 to 9.5 × 6.0 × 4.0 cm3 . Defect sizes ranged from 16 × 10 to 8 × 4 cm2 . All patients underwent multiple imaging examinations to exclude abnormalities. Primary outcomes included complete tumor resection and flap survival. Secondary outcomes included the BREAST-Q results and bilateral breast symmetry measurements. RESULTS: All flaps exhibited good postoperative survival and were between 20 × 12 and 10 × 5 cm2 . Two patients experienced minor complications that did not influence outcomes. No obvious donor site complications were observed. The BREAST-Q results indicated favorable reconstructive efficacy, with "satisfaction with breasts," "physical well-being (chest)," and "satisfaction with the outcome" scores of 77.8 (range: 58-87), 83.2 (range: 77-89), and 80.1 (range: 70-88), respectively. No signs of tumor recurrence were observed during a median follow-up period of 53.5 months (range: 6-83 months). However, poor postoperative bilateral breast symmetry was observed (vertical extent: 0.63 [range: 0.36-0.88]; horizontal extent: 0.64 [range: 0.41-0.80]). CONCLUSIONS: IMAP flaps are reliable options for chest wall defect repair and breast reconstruction for selected patients with locally advanced breast cancer and inner breast tumors. Despite poor postoperative bilateral breast symmetry, most patients reported excellent outcomes.

Micropterus salmoides rhabdovirus enters cells via clathrin-mediated endocytosis pathway in a pH-, dynamin-, microtubule-, rab5-, and rab7-dependent manner.

IMPORTANCE: Although Micropterus salmoides rhabdovirus (MSRV) causes serious fish epidemics worldwide, the detailed mechanism of MSRV entry into host cells remains unknown. Here, we comprehensively investigated the mechanism of MSRV entry into epithelioma papulosum cyprinid (EPC) cells. This study demonstrated that MSRV enters EPC cells via a low pH, dynamin-dependent, microtubule-dependent, and clathrin-mediated endocytosis. Subsequently, MSRV transports from early endosomes to late endosomes and further into lysosomes in a microtubule-dependent manner. The characterization of MSRV entry will further advance the understanding of rhabdovirus cellular entry pathways and provide novel targets for antiviral drug against MSRV infection.

Treatment options for digital nerve injury: a systematic review and meta-analysis.

BACKGROUND: Surgical treatment of finger nerve injury is common for hand trauma. However, there are various surgical options with different functional outcomes. The aims of this study are to compare the outcomes of various finger nerve surgeries and to identify factors associated with the postsurgical outcomes via a systematic review and meta-analysis. METHODS: The literature related to digital nerve repairs were retrieved comprehensively by searching the online databases of PubMed from January 1, 1965, to August 31, 2021. Data extraction, assessment of bias risk and the quality evaluation were then performed. Meta-analysis was performed using the postoperative static 2-point discrimination (S2PD) value, moving 2-point discrimination (M2PD) value, and Semmes-Weinstein monofilament testing (SWMF) good rate, modified Highet classification of nerve recovery good rate. Statistical analysis was performed using the R (V.3.6.3) software. The random effects model was used for the analysis. A systematic review was also performed on the other influencing factors especially the type of injury and postoperative complications of digital nerve repair. RESULTS: Sixty-six studies with 2446 cases were included in this study. The polyglycolic acid conduit group has the best S2PD value (6.71 mm), while the neurorrhaphy group has the best M2PD value (4.91 mm). End-to-side coaptation has the highest modified Highet's scoring (98%), and autologous nerve graft has the highest SWMF (91%). Age, the size of the gap, and the type of injury were factors that may affect recovery. The type of injury has an impact on the postoperative outcome of neurorrhaphy. Complications reported in the studies were mainly neuroma, cold sensitivity, paresthesia, postoperative infection, and pain. CONCLUSION: Our study demonstrated that the results of surgical treatment of digital nerve injury are generally satisfactory; however, no nerve repair method has absolute advantages. When choosing a surgical approach to repair finger nerve injury, we must comprehensively consider various factors, especially the gap size of the nerve defect, and postoperative complications. Type of study/level of evidence Therapeutic IV.

The ABI5-dependent down-regulation of mitochondrial ATP synthase OSCP subunit facilitates apple necrotic mosaic virus infection.

Apple necrotic mosaic virus (ApNMV) is associated with apple mosaic disease in China. However, the mechanisms of ApNMV infection, as well as host defence against the virus, are still poorly understood. Mitochondrial ATP synthase plays a fundamental role in the regulation of plant growth and development. However, mitochondrial ATP synthase function in response to virus infection remains to be defined. In the present study, a yeast two-hybrid (Y2H) screening revealed that the apple mitochondrial ATP synthase oligomycin sensitivity-conferring protein (OSCP) subunit (MdATPO) interacts with ApNMV coat protein (CP). It was further verified that overexpression of MdATPO in Nicotiana benthamiana inhibited viral accumulation. In contrast, silencing of NbATPO facilitated viral accumulation, indicating that ATPO plays a defensive role during ApNMV infection. Further investigation demonstrated that ApNMV infection accelerated abscisic acid (ABA) accumulation, and ABA negatively regulated ATPO transcription, which was related to the ability of ABA insensitive 5 (ABI5) to bind to the ABA-responsive elements (ABREs) of the ATPO promoter. Taken together, our results indicated that transcription factor ABI5 negatively regulated ATPO transcription by directly binding to its promoter, leading to the susceptibility of apple and N. benthamiana to ApNMV infection. The current study facilitates a comprehensive understanding of the intricate responses of the host to ApNMV infection.

TRIM72 Alleviates Muscle Inflammation in mdx Mice via Promoting Mitophagy-Mediated NLRP3 Inflammasome Inactivation.

Chronic muscle inflammation exacerbates the pathogenesis of Duchenne muscular dystrophy (DMD), which is characterized by progressive muscle degeneration and weakness. NLRP3 (nucleotide-binding domain and leucine-rich repeat pyrin domain containing 3) inflammasome plays a key role in the inflammatory process, and its abnormal activation leads to a variety of inflammatory or immune diseases. TRIM72 (MG53) is a protective myokine for tissue repair and regeneration. However, little is known about the potential impact of TRIM72 in the crosstalk between mitophagy and inflammatory process of DMD. Here, 10-week-old male mdx mice were injected intramuscularly with adeno-associated virus (AAV-TRIM72) to overexpress TRIM72 protein for 6 weeks. Then, skeletal muscle samples were collected, and relevant parameters were measured by histopathological analysis and molecular biology techniques. C2C12 cell line was transfected with lentivirus (LV-TRIM72) to overexpress or siRNA (si-TRIM72) to suppress the TRIM72 expression for the following experiment. Our data firstly showed that the TRIM72 expression was decreased in skeletal muscles of mdx mice. Then, we observed the increased NLRP3 inflammasome and impaired mitophagy in mdx mice compared with wild type mice. In mdx mice, administration of AAV-TRIM72 alleviated the accumulation of NLRP3 inflammasome and the consequent IL-18 and IL1ß maturation by inducing autophagy, while this protective effect was reversed by chloroquine. Mitochondrial reactive oxygen species (mtROS), as a recognized activator for NLRP3 inflammasome, was attenuated by TRIM72 through the induction of mitophagy in C2C12 cells. Additionally, we proposed that the TRIM72 overexpression might promote mitophagy through both the early stage by PI3K-AKT pathway and the late stage by autolysosome fusion. In conclusion, the current study suggests that TRIM72 prevents DMD inflammation via decreasing NLRP3 inflammasomes and enhancing mitophagy. Collectively, our study provides insight into TRIM72 as a promising target for therapeutic intervention for DMD.

Research on health risk assessment of heavy metals in soil based on multi-factor source apportionment: A case study in Guangdong Province, China.

Environmental problems caused by heavy metal pollution in soil have attracted widespread attention worldwide. Identifying and quantifying the heavy metal pollution sources and risks is crucial for subsequent soil management. In this study, an integrated source-risk method for source apportionment and risk assessment based on the PMF model, the geodetector model and the health risk assessment model (HRA) was proposed and applied. Analysis of Hg, As, Pb, Cd, Cu, Ni, Cr, and Zn in 208 topsoils showed that the average contents of eight heavy metals were 1.87-5.86 times greater than corresponding background values, among which Cd and As were relatively high, which were higher than the specified soil risk screening values, high-value areas of heavy metals are mainly concentrated in the central part of the study area. The source apportionment showed that the accumulation of heavy metals was affected by five sources: atmospheric deposition (16.3 %), natural sources (33.1 %), industrial activities dominated by metal mining (15.1 %), industrial activities dominated by metal smelting (12.6 %) and traffic sources (22.9 %). The results of the health risk assessment showed that the carcinogenic risks (adult: 4.74E-05, children: 7.41E-05) of heavy metals in soil to the study population were both acceptable, the non-carcinogenic risk of adult (THI = 0.277) was within the limit, while the non-carcinogenic risk of children (THI = 1.70) was higher than the limit value. Ingestion (89.5 %-95.9 %) contributed the greatest health risk among all exposure routes. Source 3 (arsenic-related industrial activities dominated by metal mining) contributed the most to the HI and CRI of adults and children (all above 50 %), therefore, in the formulation stage of soil management strategy in this area, priority should be given to the control and management of this pollution source. These results can provide more detailed support for environmental protection departments to propose targeted soil pollution control measures.

Development and validation of a predictive nomogram for subsequent contralateral hip fracture in elderly patients within 2 years after hip fracture surgery.

Purpose: Contralateral hip refracture following initial hip fracture surgery is life-threatening in the elderly with high incidence and mortality. This study investigated the associated independent risk factors and established a nomogram prediction model. Methods: Totally 734 elderly patients with hip fractures who underwent surgical treatment (January 2016-December 2020) were enrolled. Following analyses on demographic variables, clinical characteristics, and laboratory examination, independent risk factors of contralateral hip fractures in the elderly were identified through the least absolute shrinkage and selection operator (LASSO) regression, and univariate and multivariate logistic regression. Patients were randomly allocated into training (n = 513) and validation sets (n = 221). A training set-based nomogram prediction model was established and assessed for predictability, discriminatory ability, and clinical applicability using the receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA) in both sets. Results: Contralateral hip refractures occurred in 7.08% (52/734) patients within 2 years after surgery. Age, hemoglobin (Hb), heart disease, neurovascular disease, Parkinson's disease (PD), Alzheimer's disease (AD), chronic obstructive pulmonary disease (COPD), and chronic kidney disease (CKD) were independent risk factors. The nomogram prediction model had a favorable discriminatory ability, as indicated by the areas under the ROC curves (AUC): 0.906 (95% CI, 0.845-0.967) in the training set and 0.956 (95% CI, 0.927-0.985) in the validation set. The calibration curves demonstrated a good consistency between the actual subsequent contralateral hip fracture incidence and the predicted probability. The DCA of the nomogram demonstrated the model's excellent clinical efficacy. Conclusion: The nomogram model enabled accurate individualized prediction for the occurrence of subsequent contralateral hip fracture in the elderly within 2 years after surgical treatment, which might help clinicians with precise references for appropriate perioperative management and rehabilitation education following initial hip surgery for their patients.