Prognostic Value of Lymphocyte-to-Monocyte Ratio (LMR) in Patients With Prostate Cancer: A Systematic Review and Meta-Analysis.

The objective of this study is to evaluate the prognostic value of lymphocyte-to-monocyte ratio (LMR) in patients with prostate cancer (PCa) by a method of meta-analysis. China National Knowledge Infrastructure (CNKI), Wanfang Data, PubMed, Web of Science, Cochrane Library, and Embase were searched to collect relevant literature until March 2023. The Newcastle-Ottawa Scale was used to assess the bias risk of the literature included. Hazard ratios (HRs) and 95% confidence intervals (CIs) were used to evaluate the prognostic value of LMR in PCa. Stata 15.0 statistical software was used for data analysis. A total of six published articles were included in this meta-analysis, containing 1,104 patients with PCa. The results of the meta-analysis indicated better overall survival (OS; HR = 1.73, 95% CI: 1.73, p = .001) and progression-free survival (PFS; HR = 2.63, 95% CI: 1.58~4.38, p < .001) in patients with PCa with low LMR compared with high LMR. In conclusion, compared with low LMR, PCa patients with high LMR have a better prognosis. LMR is an independent risk factor affecting the long-term prognosis of patients with PCa. The detection of LMR before treatment is of certain significance in judging the clinical prognosis of patients with PCa.

Engineering and Characterization of an Artificial Drug-Carrying Vesicles Nanoplatform for Enhanced Specifically Targeted Therapy of Glioblastoma.

Glioblastoma multiforme (GBM) treatment is hindered by complex pathologies and the need to cross the blood-brain barrier (BBB) during drug delivery. Although exosomes have great potential for GBM treatment, these alone cannot fully meet the therapeutic requirements, owing to their limitations in targeting and delivery. Herein, engineered artificial vesicles (EAVs), ANG-TRP-PK1@EAVs, which are constructed using a liposome extruder from HEK293T cells expressing ANG-TRP-PK1 peptides, is developed. ANG-TRP-PK1 is a fusion peptide of Angiopep-2 fused to the N-terminus of TRP-PK1, to present Angiopep-2 on the EAVs. ANG-TRP-PK1@EAVs have similar characteristics to the secreted exosomes, but a much higher yield. ANG-TRP-PK1@EAVs have efficient BBB-penetration and GBM-targeting abilities in a mock BBB model in in vitro and orthotopic GBM mouse models in vivo. Doxorubicin loading EAVs (ANG-TRP-PK1@DOX) do not alter the characteristics of the EAVs, which can cross the BBB, reach the GBM, and kill tumor cells in orthotopic GBM mouse models. These engineered drug-loaded artificial vesicles show better therapeutic effects on GBM than temozolomide in mice, with very few side effects. In conclusion, EAVs can be inserted into different targeting ligands and packed into different drugs, and they may serve as unique and efficient nanoplatforms for drug delivery and tumor promise therapy.

Investigation of Diagnostic and Prognostic Value of CLEC4M of Non-Small Cell Lung Carcinoma Associated with Immune Microenvironment.

Purpose: C-type lectin domain family 4 member M (CLEC4M) has been found to be involved in the occurrence and development of cancer, but its role in NSCLC remains to be fully explored. Our work aims to evaluate the diagnostic and prognostic value of CLEC4M in NSCLC and to investigate the underlying mechanisms of CLEC4M in the immune microenvironment of NSCLC. Methods: Integrating publicly accessible data and clinical tissue samples to verify the expression of CLEC4M in NSCLC. The diagnostic value of CLEC4M was determined by receiver operating characteristic (ROC) curve. Kaplan-Meier survival analysis, nomogram plot, univariate and multivariate Cox regression models were performed to evaluate the prognostic impact of CLEC4M on NSCLC patients. The correlation between CLEC4M and tumor immune infiltration was estimated using TIMER and UALCAN databases. Functional assessments including GO, KEGG pathway and GSEA analyses were implemented to illustrate the potential mechanisms of CLEC4M in NSCLC. Results: CLEC4M was significantly downregulated in NSCLC tissue, as confirmed by immunohistochemistry of clinical tissues. The high AUC value of ROC curves demonstrated the diagnostic accuracy of CLEC4M in NSCLC. Additionally, low CLEC4M expression was associated with poor survival in NSCLC patients. Furthermore, CLEC4M was found to be significantly associated with tumor immune infiltration, and CLEC4M may be involved in immune activation and proliferation inhibition through the functional assessment, suggesting that CLEC4M may be a therapeutic target for NSCLC patients. Conclusion: Our findings reveal CLEC4M is significantly downregulated in NSCLC tissues, and illustrate the diagnostic and prognostic value of CLEC4M in NSCLC, as well as its potential serve as an immune-related therapeutic target.

Novel Predictors of Stroke-Associated Pneumonia: A Single Center Analysis.

Stroke-associated pneumonia (SAP) is a common cause of disability or death. Although the researches on SAP have been relatively mature, the method that can predict SAP with great accuracy has not yet been determined. It is necessary to discover new predictors to construct a more accurate predictive model for SAP. We continuously collected 2,366 patients with acute ischemic stroke, and then divided them into the SAP group and non-SAP group. Data were recorded at admission. Through univariate analyses and multivariate regression analyses of the data, the new predictive factors and the predictive model of SAP were determined. The receiver operating characteristic (ROC) curve and the corresponding area under the curve (AUC) were used to measure their predictive accuracy. Of the 2,366 patients, 459 were diagnosed with SAP. International normalized ratio (INR) (odds ratio = 37.981; 95% confidence interval, 7.487-192.665; P < 0.001), age and dysphagia were independent risk factors of SAP. However, walking ability within 48 h of admission (WA) (odds ratio = 0.395; 95% confidence interval, 0.287-0.543; P < 0.001) was a protective factor of SAP. Different predictors and the predictive model all could predict SAP (P < 0.001). The predictive power of the model (AUC: 0.851) which included age, homocysteine, INR, history of chronic obstructive pulmonary disease (COPD), dysphagia, and WA was greater than that of age (AUC: 0.738) and INR (AUC: 0.685). Finally, we found that a higher INR and no WA could predict SAP in patients with acute ischemic stroke. In addition, we designed a simple and practical predictive model for SAP, which showed relatively good accuracy. These findings might help identify high-risk patients with SAP and provide a reference for the timely use of preventive antibiotics.

Use of latent profile analysis and k-means clustering to identify student anxiety profiles.

BACKGROUND: Anxiety disorders are often the first presentation of psychopathology in youth and are considered the most common psychiatric disorders in children and adolescents. This study aimed to identify distinct student anxiety profiles to develop targeted interventions. METHODS: A cross-sectional study was conducted with 9738 students in Yingshan County. Background characteristics were collected and Mental Health Test (MHT) were completed. Latent profile analysis (LPA) was applied to define student anxiety profiles, and then the analysis was repeated using k-means clustering. RESULTS: LPA yielded 3 profiles: the low-risk, mild-risk and high-risk groups, which comprised 29.5, 38.1 and 32.4% of the sample, respectively. Repeating the analysis using k-means clustering resulted in similar groupings. Most students in a particular k-means cluster were primarily in a single LPA-derived student profile. The multinomial ordinal logistic regression results showed that the high-risk group was more likely to be female, junior, and introverted, to live in a town, to have lower or average academic performance, to have heavy or average academic pressure, and to be in schools that have never or occasionally have organized mental health education activities. CONCLUSIONS: The findings suggest that students with anxiety symptoms may be categorized into distinct profiles that are amenable to varying strategies for coordinated interventions.

Neurog2 directly converts astrocytes into functional neurons in midbrain and spinal cord.

Conversion of astrocytes into neurons in vivo offers an alternative therapeutic approach for neuronal loss after injury or disease. However, not only the efficiency of the conversion of astrocytes into functional neurons by single Neurog2, but also the conundrum that whether Neurog2-induced neuronal cells (Neurog2-iNs) are further functionally integrated into existing matured neural circuits remains unknown. Here, we adopted the AAV(2/8) delivery system to overexpress single factor Neurog2 into astrocytes and found that the majority of astrocytes were successfully converted into neuronal cells in multiple brain regions, including the midbrain and spinal cord. In the midbrain, Neurog2-induced neuronal cells (Neurog2-iNs) exhibit neuronal morphology, mature electrophysiological properties, glutamatergic identity (about 60%), and synapse-like configuration local circuits. In the spinal cord, astrocytes from both the intact and lesioned sources could be converted into functional neurons with ectopic expression of Neurog2 alone. Notably, further evidence from our study also proves that Neurog2-iNs in the intact spinal cord are capable of responding to diverse afferent inputs from dorsal root ganglion (DRG). Together, this study does not merely demonstrate the feasibility of Neurog2 for efficient in vivo reprogramming, it gives an indication for the Neurog2-iNs as a functional and potential factor in cell-replacement therapy.

Conversion of Astrocytes and Fibroblasts into Functional Noradrenergic Neurons.

Dysfunction of noradrenergic (NA) neurons is associated with a number of neuronal disorders. Diverse neuronal subtypes can be generated by direct reprogramming. However, it is still unknown how to convert non-neuronal cells into NA neurons. Here, we show that seven transcription factors (TFs) (Ascl1, Phox2b, AP-2α, Gata3, Hand2, Nurr1, and Phox2a) are able to convert astrocytes and fibroblasts into induced NA (iNA) neurons. These iNA neurons express the genes required for the biosynthesis, release, and re-uptake of noradrenaline. Moreover, iNA neurons fire action potentials, receive synaptic inputs, and control the beating rate of co-cultured ventricular myocytes. Furthermore, iNA neurons survive and integrate into neural circuits after transplantation. Last, human fibroblasts can be converted into functional iNA neurons as well. Together, iNA neurons are generated by direct reprogramming, and they could be potentially useful for disease modeling and cell-based therapies.

APPBP2 enhances non-small cell lung cancer proliferation and invasiveness through regulating PPM1D and SPOP.

BACKGROUND: The influence of amyloid protein-binding protein 2 (APPBP2) on lung cancer is unknown. METHODS: The function and mechanisms of APPBP2 were investigated in the NSCLC cell lines A549 and H1299. The ectopic expression of APPBP2, PPM1D and SPOP in NSCLS were examined in samples collected from ten pairs of human lung adenocarcinoma cancer tissues and adjacent normal lung tissues. shRNA vector was used for APPBP2 knockdown. Quantitative PCR and western blot assays quantified the mRNA and protein level of APPBP2, PPM1D, and SPOP. Cell proliferation was measured with BrdU, MTT, colony formation assays, and xenograft tumour growth experiments. Cell migration and invasion were analysed with transwell and wound healing assays. Co-Immunoprecipitation assay detected protein-protein interactions. FINDINGS: APPBP2 was upregulated in NSCLC tissues. Silencing APPBP2 in A549 and H1299 cells resulted in the inhibition of cell proliferation, migration, and invasion, enhancement of apoptosis, and a significant decrease in the expression of PPM1D and SPOP. Overexpression of PPM1D and SPOP attenuated the APPBP2-knockdown inhibition of NSCLC cells. Co-IP assay showed that PPM1D interacted with APPBP2. INTERPRETATION: The expression level of APPBP2 positively correlates with NSCLC cell proliferation, migration, and invasiveness. APPBP2 contributes to NSCLC progression through regulating the PPM1D and SPOP signalling pathway. This novel molecular mechanism, underlying NSCLC oncogenesis, suggests APPBP2 is a potential target for diagnosis and therapeutic intervention in NSCLC. FUND: Key Program of Natural Science Research of Higher Education of Anhui Province (No. KJ2017A241), the National Natural Science Foundation of China (No. 81772493).

Ascl1 Converts Dorsal Midbrain Astrocytes into Functional Neurons In Vivo.

In vivo induction of non-neuronal cells into neurons by transcription factors offers potential therapeutic approaches for neural regeneration. Although generation of induced neuronal (iN) cells in vitro and in vivo has been reported, whether iN cells can be fully integrated into existing circuits remains unclear. Here we show that expression of achaete-scute complex homolog-like 1 (Ascl1) alone is sufficient to convert dorsal midbrain astrocytes of mice into functional iN cells in vitro and in vivo. Specific expression of Ascl1 in astrocytes by infection with GFAP-adeno-associated virus (AAV) vector converts astrocytes in dorsal midbrain, striatum, and somatosensory cortex of postnatal and adult mice into functional neurons in vivo. These iN cells mature progressively, exhibiting neuronal morphology and markers, action potentials, and synaptic inputs from and output to existing neurons. Thus, a single transcription factor, Ascl1, is sufficient to convert brain astrocytes into functional neurons, and GFAP-AAV is an efficient vector for generating iN cells from astrocytes in vivo.

[Effect of electroacupuncture intervention on N-methyl-D-aspartic acid receptor expression in spinal cord in rats with chronic constrictive injury of the sciatic nerve].

UNLABELLED: OBJECTIVE; To observe the changes of N-methyl-D-aspartic acid (NMDA) receptor expression of spinal cord after electroacupuncture (EA) intervention in rats with chronic constrictive injury (CCI) of the sciatic nerve so as to reveal the mechanism underlying improvement of neuropathic pain. METHODS: Sixty male SD rats were randomly divided into sham operation (sham) group, CCI model group and EA group (n = 20). CCI model was established by ligature of the right sciatic nerve with a piece of surgical chromic suture. For rats of the sham group, the right sciatic nerve was just isolated without ligature. The rats of the EA group were given with EA stimulation of "Weizhong" (BL 40) and "Huantiao" (GB 30) on the injured side at a frequency of 2 Hz, electric current of 1-3 mA for 30 min (increasing 1 mA every 10 min). The treatment was conducted once a day from the 11th day to the 20th day after modeling. NMDA receptor 2 B subunit (NR 2 B) protein and mRNA expression levels in the spinal cord were determined by immunohistochemistry, Western blot and reverse transcription (RT)-polymerase chain reaction (PCR), respectively, and spinal NMDA receptor subunit 1 (NR 1) protein and mRNA expression levels were measured by Western blot and RT-PCR, respectively. RESULTS: In comparison with the sham group, NR 1 protein and mRNA expression levels of the model group in the spinal cord were considerably upregulated after CCI (P < 0.01, P < 0.05). In comparison with the model group NR 1 protein and mRNA expression levels of the EA group in the spinal cord were evidently down-regulated (P < 0.05). No significant changes of NR 2 B protein and mRNA expression after CCI and EA stimulation were found after immunohistochemistry, Western blot and RT-PCR measurements (P > 0.05). CONCLUSION: EA intervention is effective in alleviating neuropathic pain in CCI rats, which may be closely related to its effects in lowering functional activity of NR 1 protein and mRNA in the spinal cord.

MicroRNAs modulate the Wnt signaling pathway through targeting its inhibitors.

Increasing evidence indicates that microRNAs (miRNAs) play important roles in mouse brain development. We and several other reports recently have demonstrated that Wnt1-cre-mediated loss of Dicer, the key enzyme for miRNA biosynthesis, results in malformation of the midbrain and cerebellum and failure of neural crest and dopaminergic differentiation. The underlying mechanisms, however, remain poorly understood. The resemblance of some of the phenotypes in the Wnt1-cre Dicer conditional knockout embryos and Wnt1(-/-), Wnt1(-/-);Wnt3(-/-) and Wnt1-cre;ß-catenin(flox/flox) knockout embryos reminds us that loss of miRNA may disrupt the Wnt-ß-catenin signaling. Here we provide evidence that miRNAs modulate the Wnt signaling pathway through targeting its inhibitors. First, we predicted miRNA binding sites in the 3' UTRs of candidate inhibitors of the Wnt signaling pathway and luciferase assays revealed that several inhibitors of Wnt signaling pathway were targeted by miRNAs. Second, we demonstrated that several miRNAs could modulate the expression of Gsk3b, an inhibitor of Wnt signaling, post-transcriptional in 293T cells. Third, we found that several miRNAs were able to regulate the Wnt-ß-catenin signaling activity in 293T cells. More interestingly, the expression of ß-catenin protein was dramatically reduced in the Wnt1-cre-meidiated Dicer knockout brain tissue compared with control. Our studies therefore suggest that miRNAs might exert their functions, at least in part, by modulating the Wnt signaling pathway through targeting its inhibitors.

FoxO1 regulates muscle fiber-type specification and inhibits calcineurin signaling during C2C12 myoblast differentiation.

Adult skeletal muscle fibers can be categorized into slow-oxidative and fast-glycolytic subtypes based on specialized metabolic and contractile properties. The Forkhead box O1 (FoxO1) transcription factor governs muscle growth, metabolism, and cell differentiation, and has been shown to be involved in regulating muscle fiber type specification. However, to date, the mechanism behind FoxO1-mediated fiber type diversity is still unclear. In this article, FoxO1 being expressed preferentially in fast twitch fiber enriched muscles is reported. Moreover, the autors also detected that FoxO1 expression decreased in both fast and slow muscles from mice undergoing endurance exercise which induced a fast-to-slow fiber type transition. Using C2C12 myoblast, constitutively active FoxO1 mutant altered the proportion of muscle fiber type composition toward a fast-glycolytic phenotype and attenuated calcineurin phosphatase activity. In addition, a transcriptionally inactive FoxO1 by resveratrol triggered the expression of genes related to slow-oxidative muscle but not sufficient to induce a complete slow fiber transformation. Taken together, these results suggest that FoxO1 up-regulates fast fiber-type formation and down-regulates muscle oxidative capacity at least in part through inhibition of the calcineurin pathway.

Wnt1-cre-mediated conditional loss of Dicer results in malformation of the midbrain and cerebellum and failure of neural crest and dopaminergic differentiation in mice.

The involvement of microRNAs (miRNAs) in the development of the neural crest (NC) cells and other neuronal differentiation is still poorly understood. Here, we investigated the global function of miRNAs in embryonic development by examining the Wnt1-cre-mediated Dicer knockout mice. Dicer ablation resulted in malformation of the midbrain and cerebellum and failure of NC and dopaminergic differentiation. First, the Dicer mutant fetuses exhibited dramatic malformation of the tectum and cerebellum and the eyelids were open. Second, the skeletal structures that are derived from the cranial NC were lost or mostly ablated in Dicer mutant mice. Third, deletion of Dicer in the NC cells resulted in the malformation of the dorsal root ganglia, enteric nervous system and sympathetic ganglia. Interestingly, the expression of neuropeptide Y and its potential regulators TrkA, AP-2alpha and AP-2beta was largely abolished in sympathetic neurons of Dicer mutant mice. Fourth, in situ hybridization data revealed that the expression of miR-9, miR-124 and miR-218 in the midbrain and rostral hindbrain area was mostly eliminated in the Dicer mutant mice. We then demonstrated that the development of dopaminergic neurons was impaired in Dicer-deleted mice. Our studies therefore suggest that miRNAs contribute to the embryonic development in multiple locations.

[Over-expression of FoxO1 inhibits the differentiation of porcine skeletal muscle myoblast].

The Forkhead box O1 (FoxO1) transcription factor governs muscle growth, metabolism and cell differentiation. However, its role in myoblast differentiation is unclear. To study the biological function of FoxO1 during differentiation in porcine primary myoblast, we constructed stably FoxO1 over-expressed porcine myoblast mediated by liposome and adopted morphological observation, quantitative real-time RT-PCR and Western blotting methods to analyze FoxO1 and early and late myogenic regulation factors MyoD and myogenin expression. During differentiation the mRNA level of FoxO1 was significantly increased. However, the total protein did not change but the phosphorylation of FoxO1 was upregulated. Furthermore, overexpression of FoxO1 in porcine myoblast decreased MyoD and myogenin mRNA, whereas MyoD protein changed little and myogenin was significantly suppressed (P < 0.05). These results indicated that FoxO1 delays and negatively regulates the porcine myoblast differentiation. Moreover, FoxO1 may play a critical role in muscle fiber-type specification through the inhibition of myogenic regulation factors.

[Effects of heavenly stem-prescription of point selection of needling methods of Ziwu Liuzhu on ischemic cerebrovascular diseases].

OBJECTIVE: To observe effects of heavenly stem-prescription of point selection of the needling methods of midnight-noon ebb-flow on motorial and neurological functional deficit of ischemic cerebrovascular diseases. METHODS: The patients were randomly divided into a Ziwu Liuzhu group and a channel acupoint selection group. In the channel acupoint selection group, Jianyu (LI 15), Quchi (LI 11), Waiguan (TE 5), Zusanli (ST 36) were selected, combined with selected acupoints by syndrome differentiation. In the Ziwu Liuzhu group, on the basis of the channel acupoint selection group, according to the principle "the acupoints of the yang-channel were opened at yang-day-yang-hour and the acupoints of the yin-channel were opened at yin-day-yin-hour", and acupoints were selected by the day-hour acupoint-opening method of the Ziwu Liuzhu Najia method in Zhenjiu Daquan. The patients in the both groups began to be treated at the Chen hour (7:00-9:00) or Si hour (9:00-11:00) in the morning, 10 times constituting one course. They were treated for 3 courses. Improvement of the motor function symptom cumulative score, Fugl-Meyer (FMA) and improvement of neurological functional deficit were investigated. RESULTS: The FMA score (36.13 +/- 21.80) after treatment was significantly lower than (73.50 +/- 21.53) before treatment (P<0.01) in the Ziwu Liuzhu group, and (54.43 +/- 20.89) after treatment was significantly lower than (62.27 +/- 22.91) before treatment (P<0.05) in the channel acupoint selection group; the neurological functional deficit score (15.40 +/- 9.34) after treatment was significantly lower than (27.17 +/- 10.81) before treatment in the Ziwu Liuzhu group, and (23.97 +/- 1.30) was lower than (27.97 +/- 7.72) before treatment in the channel acupoint selection group. After treatment, FMA and the neurological functional scores significantly improved in the Ziwu Liuzhu group as compared with those in the channel acupoint selection group (P<0.05). CONCLUSION: Ziwu Liuzhu Najia point-selection method can effectively improve both nervous function and motor function in the patient of ischemic cerebrovascular disease.

Effect of acupuncture on the auditory evoked brain stem potential in Parkinson's disease.

Under the auditory evoked brain stem potential (ABP) examination, the latent period of V wave and the intermittent periods of III-V peak and I-V peak were significantly shortened in Parkinson's disease patients of the treatment group (N = 29) after acupuncture treatment. The difference of cumulative scores in Webster's scale was also decreased in correlation analysis. The increase of dopamine in the brain and the excitability of the dopamine neurons may contribute to the therapeutic effects, in TCM terms, of subduing the pathogenic wind and tranquilizing the mind.