Protein phosphatase 2Cm-regulated branched-chain amino acid catabolic defect in dorsal root ganglion neurons drives pain sensitization.

Maladaptive changes of metabolic patterns in the lumbar dorsal root ganglion (DRG) are critical for nociceptive hypersensitivity genesis. The accumulation of branched-chain amino acids (BCAAs) in DRG has been implicated in mechanical allodynia and thermal hyperalgesia, but the exact mechanism is not fully understood. This study aimed to explore how BCAA catabolism in DRG modulates pain sensitization. Wildtype male mice were fed a high-fat diet (HFD) for 8 weeks. Adult PP2Cmfl/fl mice of both sexes were intrathecally injected with pAAV9-hSyn-Cre to delete the mitochondrial targeted 2 C-type serine/threonine protein phosphatase (PP2Cm) in DRG neurons. Here, we reported that BCAA catabolism was impaired in the lumbar 4-5 (L4-L5) DRGs of mice fed a high-fat diet (HFD). Conditional deletion of PP2Cm in DRG neurons led to mechanical allodynia, heat and cold hyperalgesia. Mechanistically, the genetic knockout of PP2Cm resulted in the upregulation of C-C chemokine ligand 5/C-C chemokine receptor 5 (CCL5/CCR5) axis and an increase in transient receptor potential ankyrin 1 (TRPA1) expression. Blocking the CCL5/CCR5 signaling or TRPA1 alleviated pain behaviors induced by PP2Cm deletion. Thus, targeting BCAA catabolism in DRG neurons may be a potential management strategy for pain sensitization.

Defective branched-chain amino acid catabolism in dorsal root ganglia contributes to mechanical pain.

Impaired branched-chain amino acid (BCAA) catabolism has recently been implicated in the development of mechanical pain, but the underlying molecular mechanisms are unclear. Here, we report that defective BCAA catabolism in dorsal root ganglion (DRG) neurons sensitizes mice to mechanical pain by increasing lactate production and expression of the mechanotransduction channel Piezo2. In high-fat diet-fed obese mice, we observed the downregulation of PP2Cm, a key regulator of the BCAA catabolic pathway, in DRG neurons. Mice with conditional knockout of PP2Cm in DRG neurons exhibit mechanical allodynia under normal or SNI-induced neuropathic injury conditions. Furthermore, the VAS scores in the plasma of patients with peripheral neuropathic pain are positively correlated with BCAA contents. Mechanistically, defective BCAA catabolism in DRG neurons promotes lactate production through glycolysis, which increases H3K18la modification and drives Piezo2 expression. Inhibition of lactate production or Piezo2 silencing attenuates the pain phenotype of knockout mice in response to mechanical stimuli. Therefore, our study demonstrates a causal role of defective BCAA catabolism in mechanical pain by enhancing metabolite-mediated epigenetic regulation.

Benzene metabolite hydroquinone enhances self-renewal and proliferation of preleukemic cells through the Ppar-γ pathway.

Benzene is a known hematotoxic and leukemogenic chemical. Exposure to benzene cause inhibition of hematopoietic cells. However, the mechanism of how the hematopoietic cells inhibited by benzene undergo malignant proliferation is unknown. The cells carrying leukemia-associated fusion genes are present in healthy individuals and predispose the carriers to the development of leukemia. To identify the effects of benzene on hematopoietic cells, preleukemic bone marrow (PBM) cells derived from transgenic mice carrying the Mll-Af9 fusion gene were treated with benzene metabolite hydroquinone in serial replating of colony-forming unit (CFU) assay. RNA sequencing was further employed to identify the potential key genes that contributed to benzene-initiated self-renewal and proliferation. We found that hydroquinone induced a significant increase in colony formation in PBM cells. Peroxisome proliferator-activated receptor gamma (Ppar-γ) pathway, which plays a critical role in carcinogenesis in multiple tumors, was significantly activated after hydroquinone treatment. Notably, the increased numbers of the CFUs and total PBM cells induced by hydroquinone were significantly reduced by a specific Ppar-γ inhibitor (GW9662). These findings indicated that hydroquinone can enhance self-renewal and proliferation of preleukemic cells by activating the Ppar-γ pathway. Our results provide insight into the missing link between premalignant status and development of benzene-induced leukemia, which can be intervened and prevented.

Prognostic value of exosomal noncoding RNA in hepatocellular carcinoma: a meta-analysis.

High morbidity, recurrence and mortality make hepatocellular carcinoma (HCC) a leading cause of cancer-related burden and deaths. The lack of prognostic evaluation methods weakened the therapeutic efficacy for HCC. Exosomal noncoding RNAs (ncRNAs) play a key role in cancer development. Our meta-analysis aimed to assess the prognostic value of exosome-transferred noncoding RNAs in predicting the outcomes of patients with HCC. We obtained 16 articles from PubMed, Web of Science, Scopus, and EMBASE up to 4 November 2021. The ncRNAs were divided into three parts: microRNAs (miRNA), long noncoding RNAs (lncRNA), and circular RNAs (circRNA). In the pooled hazard ratios (HRs), upregulated miRNAs were 3.06 (95% CI = 2.51-3.73), downregulated miRNAs were 3.28 (95% CI = 2.61-4.11), lncRNAs were 3.34 (95% CI = 1.87-5.96), and circRNAs were 1.76 (95% CI = 1.36-2.14). As the results of subgroup analysis, upregulated miRNAs had a pooled HR of 3.10 (95% CI = 1.66-5.81), and the HR of downregulated miRNAs was 3.04 (95% CI = 2.17-4.28) for multivariate analysis of overall survival (OS). Meanwhile, upregulated miRNAs had a pooled HR of 2.61 (95% CI = 1.89-3.60), and the HR of downregulated miRNAs was 3.77 (95% CI = 1.11-12.73) for multivariate analysis of other endpoints. Remarkably, miR-21 has a pooled HR of 2.48 (95%CI = 1.52-4.05, I2 = 0) for disease-free survival (DFS). In conclusion, the expression of exosomal noncoding RNAs can be used to evaluate the prognosis of patients with HCC. Exosome-transferred miR-21 might serve as a potential prognostic biomarker in HCC.

Obesity by High-Fat Diet Increases Pain Sensitivity by Reprogramming Branched-Chain Amino Acid Catabolism in Dorsal Root Ganglia.

Obesity is a significant health concern as a result of poor-quality diet, for example, high-fat diet (HFD). Although multiple biological and molecular changes have been identified to contribute to HFD-induced pain susceptibility, the mechanisms are not fully understood. Here, we show that mice under 8 weeks of HFD were sensitive to mechanical and thermal stimuli, which was coupled with an accumulation of branched-chain amino acids (BCAAs) in lumbar dorsal root ganglia (DRG) due to local BCAA catabolism deficiency. This HFD-induced hyperalgesic phenotype could be exacerbated by supply of excessive BCAAs or mitigated by promotion of BCAA catabolism via BT2 treatment. In addition, our results suggested that HFD-related pain hypersensitivity was associated with a pro-inflammatory status in DRG, which could be regulated by BCAA abundance. Therefore, our study demonstrates that defective BCAA catabolism in DRG facilitates HFD-induced pain hypersensitivity by triggering inflammation. These findings not only reveal metabolic underpinnings for the pathogenesis of HFD-related hyperalgesia but also offer potential targets for developing diet-based therapy of chronic pain.

Development and external validation of a nomogram for individualized adjuvant imatinib duration for high-risk gastrointestinal stromal tumors: A multicenter retrospective cohort study.

INTRODUCTION: The main emphasis of the research about adjuvant imatinib for high-risk gastrointestinal stromal tumors (GISTs) is prolonging the treatment duration and ignores the heterogeneous that 10-year recurrence rates ranged from about 20%-100%. Thus, this study evaluated the effect of different durations of adjuvant imatinib on outcomes in high-risk GISTs to explore the feasibility of individual treatment. METHODS: We analyzed 855 high-risk GIST patients from three centers who underwent macroscopically complete resection between December 2007 and September 2020. The patients were divided into training (n =564) and two validation cohorts (n = 238 and53) based on their source. Recurrence-free survival (RFS) was the primary point. Cox multivariate analysis was used to develop the nomogram. C-index, time-dependent area under the curves, and calibration plots were used to assess the performance of the nomogram. RESULTS: Univariate analysis showed that longer adjuvant imatinib was significantly associated with better 5-year RFS (p < 0.0001). Further investigation identified that the same high-risk patients with lower tumor-associated recurrence risk benefitted little from prolonged treatment and that the recommended adjuvant imatinib duration was insufficient for those with higher recurrence risk. A nomogram for predicting 2-, 3-, and 5-year RFS based on different treatment durations and four major risk factors, namely, tumor site, size, mitotic count, and rupture status, was built and validated, with a C-index of 0.82, 0.74, and 0.70 in training and two external validation cohorts, respectively. An online dynamic nomogram was further developed for clinical applications (https://ruolinliu666.shinyapps.io/GIST/), offering predictive recurrence rates based on different treatment durations and tumor features. CONCLUSIONS: We developed a nomogram to predict the recurrence risk for high-risk patients according to tumor features and treatment durations of imatinib to help physicians on decision-making for individualized treatment duration.

Key factors affecting photoactivated fungicidal activity of sodium pheophorbide a against Pestalotiopsis neglecta.

Recently, photodynamic therapy (PDT) and photoactivated pesticides have attracted considerable research attention. In the present study, we aimed to investigate the photodynamic activity of a chlorophyllous derivative, sodium pheophorbide a (SPA), and to evaluate its potential as a photoactivated fungicide. The singlet oxygen quantum yield, the photoreaction process, the anti-photobleaching ability in sterile water (H2O), the effect of light conditions on its antifungal activity, and its stability were all investigated. SPA showed significant fungicidal activity and photostability, during which Type I and Type II photodynamic reactions occurred simultaneously on Pestalotiopsis neglecta, and the influence of Type I was slightly larger than that of Type II. In addition, light promoted the antifungal activity of SPA. In particular, the antifungal activity was enhanced with increasing light intensity, and was strongest under 8000 lx conditions. Under monochromatic light sources, antifungal activity was strongest under green light s; however, the effect of monochromatic light was not as good as that of white light. From 0 to 24 h, the antifungal effect of the SPA solution was enhanced; however, the activity of the solution began to weaken after 24 h. Furthermore, our study confirmed that the antifungal activity of SPA was stable under different temperatures, pH values, and UV irradiation durations.

Sodium pheophorbide a controls cherry tomato gray mold (Botrytis cinerea) by destroying fungal cell structure and enhancing disease resistance-related enzyme activities in fruit.

Sodium pheophorbide a (SPA) is a natural photosensitizer. The present study investigated the antifungal activity and mechanism of SPA against Botrytis cinerea in vitro and in vivo. Its inhibitory effect was studied on the spore germination and mycelial growth of B. cinerea. The effects of SPA on cell wall integrity, cell membrane permeability, and mycelial morphology of B. cinerea were also determined. Additionally, how SPA effected B. cinerea in vivo was evaluated using cherry tomato fruit. The results showed that SPA effectively inhibited the spore germination and mycelial growth of B. cinerea under light conditions (4000 lx). SPA significantly affected both cell wall integrity and cell membrane permeability (P < .05). In addition, SEM analysis suggested that B. cinerea treated with SPA (12.134 mg/mL) showed abnormal mycelial morphology, including atrophy, collapse, flattening, and mycelial wall dissolution. In vivo tests showed that SPA could increase the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) significantly (P < .05); however, SPA had no significant effect on phenylalanine ammonia lyase (PAL) activity. In short, SPA could destroy the fungal cell structure and enhance disease resistance-related enzyme activity in cherry tomatoes, thereby controlling cherry tomato gray mold.

Sodium pheophorbide a has photoactivated fungicidal activity against Pestalotiopsis neglecta.

Sodium pheophorbide a (SPA) is a natural photosensitizer. To explore its antifungal activity and mechanism, we studied its inhibitory effects on spore germination and mycelial growth of Pestalotiopsis neglecta. We used sorbitol, 2-thiobarbituric acid (TBA) and electron microscopy to determine its effects on cell wall integrity, cell membrane lipid peroxidation and mycelial morphology. Finally, the effects of SPA on enzyme activity in mycelia were determined. The results showed that SPA effectively inhibited spore germination and mycelial growth of P. neglecta under light conditions (4000 lx, 24 h). Scanning electron microscopy (SEM) revealed that SPA treatment resulted in a roughened, twisted and knotted mycelial surface and abnormal mycelial growth. SPA influenced cell wall integrity, and the content of MDA, a cell membrane lipid peroxidation product was significantly increased (P < 0.05). SPA also significantly inhibited SOD, POD and PG activity, but enhanced PPO activity (P < 0.05). In conclusion, SPA may have potential to become a biological pesticide.

MiR-221 is involved in depression by regulating Wnt2/CREB/BDNF axis in hippocampal neurons.

OBJECTIVE: The aim of this study was to investigate the mechanism of miR-221 in depression. METHODS: The molecules expressions were measured by qRT-PCR and western blot. The sucrose preference test (SPT), forced swimming test (FST) and tail suspension test (TST) were used to detect depressive-like symptoms. MTT assay and flow cytometric was used to measure the proliferation and apoptosis of hippocampal neuronal. RESULTS: MiR-221 expression in the cerebrospinal fluid and serum of major depressive disorder patients and the hippocampus of chronic unpredictable mild stress (CUMS) mice were increased, while the expression of Wnt2, p-CREB and BDNF were decreased. Additionally, silence of miR-221 increased sucrose preference of CUMS mice and shortened the immobility time of CUMS mice in SPT and FST. MiR-221 could targeted regulate Wnt2, and knockdown of Wnt2 reversed the effect of miR-221 inhibitor on the proliferation and apoptosis of hippocampal neurons and countered the promoting effect of miR-221 inhibitor on the expression of Wnt2, p-CREB and BDNF. CONCLUSION: MiR-221 could promote the development of depression by regulating Wnt2/CREB/BDNF axis.

A comparative study of magnetic resonance imaging on the gray matter and resting-state function in prodromal and first-episode schizophrenia.

It is very difficult to predict the future development possibility of schizophrenia through the clinical symptoms of the high-risk cases. Therefore, how to determine the possibility of developing into schizophrenia individuals before the onset of the diseases are particularly important. The study investigated cerebral gray matter volume differences and resting-state functional connections among patients with psychosis risk syndrome (PRS), patients with first-episode schizophrenic (FES), and healthy controls (HC), aiming to provide scientific clinical evidence for schizophrenia early identification and intervention. A total of 19 PRS patients, 18 FES patients, and 29 HC were recruited. Gray matter volume and amplitude of low-frequency fluctuation (fALFF) during resting-state functional studies were measured. Comparison of gray matter volumes showed that PRS and FES groups had common reduced gray matter volume in the right caudate. PRS and FES patients showed altered connectivity mainly in the semantic processing-related brain areas. fALFF analysis found that PRF and FES patients had significant differences in fALFF values of the brain region mainly located in the subcortical network, visual recognition network, and auditory network. In addition, PRF individuals had a higher fALFF value and a lower fALFF value in the anterior wedge of the cerebral network than the HC group. Gray matter volume loss between related brain areas might appear prior to illness onset. Similar fALFF values occurred in PRS and FES groups indicated that multiple brain regions of neuronal activity abnormalities and unconventional neural network mechanism have been existed in PRS patients.

Altered functional connectivity strength and its correlations with cognitive function in subjects with ultra-high risk for psychosis at rest.

AIMS: Evidence of altered structural and functional connectivity in the frontal-occipital network is associated with cognitive deficits in patients with schizophrenia. However, the altered patterns of functional connectivity strength (FCS) in individuals with ultra-high risk (UHR) for psychosis remain unknown. In this study, whole-brain FCS was assessed to examine the altered patterns of FCS in UHR subjects. METHODS: A total of 34 UHR subjects and 37 age- and sex-matched healthy controls were enrolled to undergo resting-state functional magnetic resonance imaging. The imaging data were analyzed using the graph theory method. RESULTS: Compared with healthy controls, UHR subjects showed significantly decreased FCS in the left middle frontal gyrus and significantly increased FCS in the left calcarine cortex. The FCS values in the left middle frontal gyrus were positively correlated to the scores of the Brief Assessments of Cognitionin Schizophrenia Symbol Coding Test (r = 0.366, P = 0.033) in the UHR subjects. A negative correlation was found between the FCS values in the left calcarine cortex and the scores of the Stroop color-naming test (r = -0.475, P = 0.016) in the UHR subjects. A combination of the FCS values in the 2 brain areas showed an accuracy of 87.32%, a sensitivity of 73.53%, and a specificity of 100% for distinguishing UHR subjects from healthy controls. CONCLUSIONS: Significantly altered FCS in the frontal-occipital network is observed in the UHR subjects. Furthermore, decreased FCS in the left middle frontal gyrus and increased FCS in the left calcarine have significant correlations with the cognitive measures of the UHR subjects and thus improve our understanding of the underlying pathophysiological mechanisms of schizophrenia. Moreover, a combination of the FCS values in the 2 brain areas can serve as a potential image marker to distinguish UHR subjects from healthy controls.

Associations among maternal pre-pregnancy body mass index, gestational weight gain and risk of autism in the Han Chinese population.

BACKGROUND: Autism is a neurodevelopmental disorder with an unclear etiology. Pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) have been suggested to play a role in the etiology of autism. The current study explores the associations among maternal pre-pregnancy BMI, GWG and the risk of autism in the Han Chinese population. METHODS: Demographic information, a basic medical history and information regarding maternal pre-pregnancy and pregnancy conditions were collected from the parents of 705 Han Chinese children with autism and 2236 unrelated typically developing children. Binary logistic regressions were conducted to calculate the odds ratio (OR) for the relationship among pre-pregnancy BMI, GWG and the occurrence of autism. The interaction between pre-pregnancy BMI and GWG was analyzed by performing stratification analyses using a logistic model. RESULTS: After adjusting for the children's gender, parental age and family annual income, excessive GWG was associated with autism risk in the entire sample (OR = 1.327, 95% CI: 1.021-1.725), whereas the relationship between maternal pre-pregnancy BMI and autism was not significant. According to the stratification analyses, excessive GWG increased the risk of autism in overweight/obese mothers (OR = 2.468, 95% CI: 1.102-5.526) but not in underweight or normal weight mothers. CONCLUSIONS: The maternal pre-pregnancy BMI might not be independently associated with autism risk. However, excessive GWG might increase the autism risk of offspring of overweight and obese mothers.

[Combined cytotoxicity mechanism of chlorpyrifos and carbofuran pesticides in vitro].

OBJECTIVE: To evaluate the single and combined effects of chlorpyrifos( CPF) and carbofuran( CF) pesticides on cell lines cultured in vitro, and clarify the pattern of joint action. METHODS: Rat pheochromocytoma( PC12 cells) were treated with different concentrations of CPF( 0, 50, 100, 200 and 400 µmol/L) and CF( 0, 25, 50, 100 and 200 µmol/L) for 12 h separately, the combined effects of two kinds of pesticides should be studied respectively in the low dose( CPF 50 µmol/L, CF 25 µmol/L) and high dose( CPF 200 µmol/L, CF 100 µmol/L) levels. After exposure, detectingacetylcholinesterase( ACh E) activity and using fluorescent probe 2', 7'-dichlorfluorescin diacetate( DCFH-DA), thiobarbituric acid( TBA) method, xanthine oxidation, 5, 5 '-dithio-bis-2-nitrobenzoic acid( DTNB) coloration to detect the intracellular reactive oxygen species( ROS) production, lipid peroxidation production malondialdehyde( MDA), activity of antioxidant enzymes superoxide dismutase( SOD) and glutathione peroxidase( GPx), respectively. RESULTS: Compared with the control group, CPF and CF could decrease the ACh E activity, induce ROS overproduction in a dose-effect way and increase the activity of SOD, GPx( P < 0. 01), but MDA content showed no significant change. Factorial ANOVA revealed that the combined effect of CPF and CF, there was no interaction at lower dose level, but interaction existed at higher dose level( P < 0. 01). The main mode of action was synergistic effect. CONCLUSION: Chlorpyrifos, carbofuran single or combined, has cytotoxicity effect. The main combined effect between chlorpyrifos and carbofuran is synergistic effect, oxidative stress damage may be one of the mechanisms.

Cognitive functioning in schizophrenia with or without diabetes.

OBJECTIVE: To determine the influence of schizophrenia patients with diabetes on cognitive function. METHODS: Altogether 78 schizophrenia patients with diabetes and 118 patients with schizophrenia were enrolled and Negative and Positive Syndrome Scale, Wechsler Memory Scale-Revised Visual Reproduction Test, Wechsler Adult Intelligence Scale-Revised Digit Symbol Test, Computerized Wisconsin Card Sorting Test,Trail Making Tests, Part A and B, and Wechsler Adult Intelligence Scale-Revised Digit Span Test were used to assess the clinical syndrome and to test the cognitive function. RESULTS: Impairment of Processing Speed, Attention/Working Memory, Executive Functioning and Visual Memory in schizophrenia patients with diabetes was significantly poorer than that of the schizophrenia patients (all P<0.05). WAIS-R Digit Symbol Test score and Trail Making Test A and B scores were associated with diabetes duration and age at diabetes onset (P<0.01). CONCLUSION: The impairment of cognitive function in schizophrenia patients with diabetes is severer than schizophrenia patients, suggesting that the prevention and management of diabetes may improve cognitive outcome in schizophrenia patients.

[The mechanism and clinical applied skills of "curettage and aspiration" technique].

OBJECTIVE: To investigate the mechanism of "curettage and aspiration" and to conclude its applied technique. METHODS: 2213 complicated abdominal cases, which operation were performed with the PMOD and its "curettage and aspiration" technique from November 1997 to May 2006, were analyzed retrospectively. Meanwhile, the successful rate of operations, the duration of operation, the blood loss of operation and the postoperative complications were statistically analyzed. RESULTS: Each operation was successfully accomplished without damage of the big blood vessels and the biliary tracts. The mean blood loss of pancreatoduodenectomy and radical gastrectomy for cancer, were respectively 105 ml and 75 ml, the mean duration was respectively 3.6 hours and 2.3 hours. As to the acute cholecystitis, the resectable rafe of gallbladder was 100%. Furthermore, no case needs blood transfusion, no massive hemorrhage and concerned damage happened when performing the operation for retroperitoneal tumor. CONCLUSION: With reasonable operational principles and applied techniques, PMOD (Peng's multifunctional operativedissector) and "curettage and aspiration" technique have demonstrated its specific superiorities in the complicated abdominal operations.