Diagnostic value and cognitive regulatory roles of long non-coding RNA UCA1 in Alzheimer's disease.

BACKGROUND: To explore the diagnostic role and potential mechanism of serum lncRNA UCA1 in Alzheimer's disease (AD). METHODS: UCA1 concentration was determined using quantitative RT-PCR. The receiver operating characteristic curve was plotted to assess the diagnostic value. Cell viability and apoptotic capacity were assessed by cell counting kit-8 (CCK-8) and flow cytometry. Water maze experiments were used to test cognitive function in mice. The target genes of UCA1 were identified with a dual luciferase reporter assay. Functional and pathway analysis of miR-342-3p target genes was determined using enrichment analysis. RESULTS: The concentration of UCA1 was elevated in the AD group and represented a diagnostic possibility of AD. The silenced UCA1 reduced the roles of Aß on viability and apoptosis of SH⁃SY5Y cells by sponging miR-342-3p. The impaired cognitive impairment was partly recovered by the knockdown of the UCA1/miR-342-3p axis. Potential targets of miR-342-3p were enriched in function and pathways related to AD progression. CONCLUSION: The UCA1/miR-342-3p axis contributed to the occurrence of AD by regulating cognitive ability.

Silencing of TRAF5 enhances necroptosis in hepatocellular carcinoma by inhibiting LTBR-mediated NF-κB signaling.

Background: Hepatocellular carcinoma (HCC) is a common malignancy with poor prognosis and high mortality. This study aimed to explore the oncogenic mechanisms of TRAF5 in HCC and provide a novel therapeutic strategy for HCC. Methods: Human HCC cell lines (HepG2, HuH7, SMMC-LM3, and Hep3B), normal adult liver epithelial cells (THLE-2), and human embryonic kidney cells (HEK293T) were utilized. Cell transfection was performed for functional investigation. qRT-PCR and western blotting were used to detect mRNA expression of TRAF5, LTBR, and NF-κB and protein expression of TRAF5, p-RIP1(S166)/RIP1, p-MLKL(S345)/MLKL, LTBR, and p-NF-κB/NF-κB. Cell viability, proliferation, migration, and invasion were evaluated using CCK-8, colony formation, wound healing, and Transwell assays. Cell survival, necrosis, and apoptosis were assessed using flow cytometry and Hoechst 33342/PI double staining. Co-immunoprecipitation and immunofluorescence were performed to determine the interaction between TRAF5 and LTBR. A xenograft model was established to validate the role of TRAF5 in HCC. Results: TRAF5 knockdown inhibited HCC cell viability, colony formation, migration, invasion, and survival but enhanced necroptosis. Additionally, TRAF5 is correlated with LTBR and TRAF5 silencing down-regulated LTBR in HCC cells. LTBR knockdown inhibited HCC cell viability, while LTBR overexpression eliminated the effects of TRAF5 deficiency on inhibiting HCC cell proliferation, migration, invasion, and survival. LTBR overexpression abolished the promotive function of TRAF5 knockdown on cell necroptosis. LTBR overexpression undid the suppressive effect of TRAF5 knockdown on NF-κB signaling in HCC cells. Moreover, TRAF5 knockdown suppressed xenograft tumor growth, inhibited cell proliferation, and promoted tumor cell apoptosis. Conclusions: TRAF5 deficiency facilitates necroptosis in HCC by suppressing LTBR-mediated NF-κB signaling.

Exploring the Target and Mechanism of Radix Paeoniae Alba on Sjogren's Syndrome.

BACKGROUND: Radix Paeoniae Alba is a traditional Chinese herbal medicine. It can accelerate salivary secretion and alleviate the dry mouth of patients with Sjogren's syndrome (SS). Although it is widely used in clinical treatment, its target and mechanism remain unclear. OBJECTIVE: This study aims to analyze the main components of Radix Paeoniae Alba, explore the target genes, and propose the possible mechanism for Radix Paeoniae Alba's acceleration of salivary secretion. METHODS: The main active components and potential targets of Radix Paeoniae Alba were searched through the TCMSP database. Efforts were made to search for the related genes of Sjogren's syndrome in OMIM and GeneCards databases. Cytoscape v3.8.0 software was used to link target genes of active components and key genes of the disease. The software Autodock vina1.1.2. was adopted to simulate the interaction between active components and target genes. Human submandibular gland (HSG) cells were used in vitro experiments to verify the results of our analysis. RESULTS: ß-Sitosterol, the main component of Radix Paeoniae Alba, may intervene in the disease through CHRM3. Molecular docking shows ß-Sitosterol has a high affinity with CHRM3, and the interaction between CHRM3 and ß-Sitosterol is the basis of biological activity. The in vitro experiments showed that ß-Sitosterol could significantly up-regulate the mRNA and protein expression levels of both CHRM3 and secretion-related genes in HSG cells. CONCLUSION: Our study shows that the chemical components of Radix Paeoniae Alba have a positive effect on the related mechanism of salivary secretion. We found that ß-Sitosterol can promote the expression of CHRM3, stimulate salivary secretion, treat Sjogren's syndrome and potentially improve its prognosis.

Dysosma versipellis Extract Inhibits Esophageal Cancer Progression through the Wnt Signaling Pathway.

OBJECTIVE: In this study, we aim to investigate the effect of Dysosma versipellis extract on biological behavior of esophageal cancer cells and its underlying mechanisms. METHODS: A total of 30 BALB/C nude mice (class SPF) were equally and randomly divided into the control group, model group, and Dysosma versipellis group. CP-C cell of esophageal cancer was subcutaneously injected into the model group as well as the Dysosma versipellis group, and the same amount of normal saline into the control group, in order to compare the tumorigenesis of nude mice of three groups. Wnt, ß-catenin, and p-GSK3ß/GSK3ß expression in tumor tissues was detected using Western blot. CP-C cells in logarithmic growth were selected and divided into 4 groups, including the control group, podophyllotoxin group, Wnt activator group, and combined group (mixture of podophyllotoxin and Wnt activator). The cell viability, apoptosis, and invasion ability, Wnt, ß-catenin, and p-GSK3ß/GSK3ß expression level of CP-C cells in each group were detected via MTT assay, flow cytometry, transwell, and Western blot, respectively. RESULTS: The tumorigenesis rates of the control group, model group, and Dysosma versipellis group were 0%, 90% (1 tumor-free mouse), and 80% (2 tumor-free mice), respectively. The tumor mass in the Dysosma versipellis group was significant less than that in the model group. Based on the results of Western blot, Wnt, ß-catenin, and p-GSK3ß/GSK3ß expression of the Dysosma versipellis group was lower than that of the control group. The in vitro viability test indicated that there was a significant difference in cell viability exhibited among four groups. Cell viability level in the 3 groups, including the combined group, blank group, and Wnt activator group, was higher than the podophyllotoxin group at each time point. In vitro apoptosis assay revealed that significant differences in cell apoptosis exhibited among four groups. Cell apoptosis rate was higher in the podophyllotoxin group compared to the remaining three groups. The Wnt activator group showed the lowest cell apoptosis rate. The in vitro invasion assay demonstrated that numbers of transmembrane cell in the 3 groups, involving the combined group, blank group, and Wnt activator group, showed a higher level than the podophyllotoxin group. The results of Western blot manifested that the podophyllotoxin group showed lower level of Wnt, ß-catenin, and p-GSK3ß/GSK3ß expression compared to the other 3 groups. CONCLUSION: Podophyllotoxin in Dysosma versipellis has an excellent antiesophageal cancer effect and is able to inhibit cell viability as well as invasion ability and promote apoptosis of esophageal cancer cells by inhibiting the Wnt signaling pathway, which could be potentially used in future clinical treatment of esophageal cancer.

Circulating Irisin Levels in Patients with Nonalcoholic Fatty Liver Disease: A Systematic Review and Meta-Analysis.

BACKGROUND AND AIMS: Previous studies have revealed the close relation of irisin with the occurrence and development of nonalcoholic fatty liver disease (NAFLD). A systematic review and meta-analysis were conducted to evaluate the association of circulating irisin levels and NAFLD. METHODS: A systematic literature search of PubMed, Embase, Cochrane Library, Clinicaltrials.gov, WANFANG, CNKI, and CBM databases was performed for relevant articles till August 2020. The weighted mean difference (WMD) values and 95% confidence intervals (CIs) were estimated to compare the case-control studies and pooled results using meta-analysis. RESULTS: The meta-analysis included 5 case-control studies with a total of 1087 people. The results revealed that the circulating irisin levels showed no significant difference between NAFLD and healthy groups (WMD = 7.51 (-12.53, 27.56) ng/ml, P > 0.05). Subgroup analysis based on races showed that the average irisin levels were higher in the NAFLD group than in the healthy group (WMD = 13.53 (0.71, 26.34) ng/ml, P < 0.05) in 4 Asian studies. Subgroup analysis based on disease severity from 3 Asian studies revealed that the average irisin levels were higher in the NAFLD group than in the healthy group (WMD = 25.1 (22.85, 27.51) ng/ml, P < 0.05 and WMD = 13.52 (22.85, 27.51) ng/ml, P < 0.05, respectively). Subgroup analysis including 3 studies from Asia suggested that the irisin levels were higher in mild NAFLD than in moderate-severe NAFLD (WMD = 11.68 (9.03, 14.32) ng/ml, P < 0.05). CONCLUSION: The average irisin levels might be higher in the NAFLD group than in the healthy group in Asians. The irisin levels in the mild NAFLD group might be higher than those in the moderate-severe group in Asians. It is important to monitor the changing trend of irisin levels in predicting the course of NAFLD disease and its changes.

Synthesis and bioevaluation of 1-phenyl-pyrazole-4-carboxylic acid derivatives as potent xanthine oxidoreductase inhibitors.

A diverse library of 1-phenyl-pyrazole-4-carboxylic acid derivatives were synthesized and evaluated for their inhibitory potency against xanthine oxidoreductase (XOR) in vitro and vivo, and the structure-activity relationship (SAR) analyses were also presented. Approximately half of the target compounds exhibited the inhibitory potency for XOR at the nanomolar level. Compounds 16c, 16d, and 16f emerged as the most potent xanthine oxidoreductase inhibitors with IC50 values of 5.7, 5.7 and 4.2 nM, respectively, in comparison to febuxostat (IC50 of 5.4 nM). Steady-state kinetics measurements indicated that 16c is a mixed-type inhibitor. A computer molecular docking study of 16c bound to XOR was performed to gain an insight into its bind mode and SAR for the series. A potassium oxonate-hypoxanthine-induced hyperuricemia model in mice was chosen to further confirm the hypouricemic effects of 16c and 16f, and the results demonstrated that 16c exhibits similar hypouricemic potency to febuxostat.

The E3 ubiquitin ligase seven in absentia homolog 1 may be a potential new therapeutic target for Parkinson's disease.

In this study, we investigated the effect of an antibody against E3 ubiquitin ligase seven in absentia homolog 1 (SIAH-1) in PC12 cells. 1-Methyl-4-phenylpyridinium (MPP(+)) treatment increased α-synuclein, E1 and SIAH-1 protein levels in PC12 cells, and it reduced cell viability; however, there was no significant change in light chain 3 expression. Treatment with an SIAH-1 antibody decreased mRNA expression levels of α-synuclein, light chain 3 and SIAH-1, but increased E1 mRNA expression. It also increased cell viability. Combined treatment with MPP(+) and rapamycin reduced SIAH-1 and α-synuclein levels. Treatment with SIAH-1 antibody alone diminished α-synuclein immunoreactivity in PC12 cells, and reduced the colocalization of α-synuclein and light chain 3. These findings suggest that the SIAH-1 antibody reduces the monoubiquitination and aggregation of α-synuclein, promoting its degradation by the ubiquitin-proteasome pathway. Consequently, SIAH-1 may be a potential new therapeutic target for Parkinson's disease.

Proxies of cognitive reserve and their effects on neuropsychological performance in patients with mild cognitive impairment.

Cognitive reserve (CR) modulates the relationship between clinical and pathological phenotypes by restricting the negative effect of cerebral lesions on cognition, according to the CR hypothesis. In this study, we evaluated 18 healthy subjects and 21 patients with mild cognitive impairment (MCI) using conventional MRI, magnetic resonance spectroscopy (MRS), the Mini-Mental State Examination (MMSE), the Wechsler Memory Scale (WMS), and the Montreal Cognitive Assessment (MoCA). The MMSE, WMS and MoCA assessments were repeated 1year later. The MRS analysis results showed that the N-acetyl-aspartate peak area (NAA; t=5.122, P<0.001) and the NAA/creatine (Cr), NAA/myo-inositol (mI) and NAA/choline (Cho) ratios were significantly changed in patients with MCI compared with the control subjects (all P<0.01). The MoCA was significantly related to the NAA/Cho ratio (R=0.443), the NAA/Cr ratio (R=0.533), and the NAA peak area (R=0.814; all P<0.05), but was unrelated to the NAA/mI ratio (R=0.400, P=0.072). We found that the hippocampal volume was significantly correlated with the MoCA, WMS and MMSE (R=0.704, 0.677, 0.542 respectively; all P<0.05). Education level had a positive effect on changes in the MoCA, MMSE, and WMS 1year later. We believe that MoCA and WMS results, MRI hippocampal volume and other related indicators (NAA peak area, NAA/Cr ratio, NAA/mI ratio, and NAA/Cho ratio) may reflect the degree of CR capacity, and that the number of years of education can significantly affect the changes in cognitive function in patients with MCI. Therefore, increasing levels of education and life skills training can help increase CR, reduce the risk of MCI, and slow down the appearance of clinical manifestations and clinical progress in patients with MCI.

Screening and determinations of tissue polypeptide antigen by label-free optical immunosensing method.

Based on the specificity of the immunoreaction of anti-body and antigen, and the resulting localized surface plasmon resonance extinction response of functionalized nano-Au monolayer on glass chip, a novel label-free optical immunosensor with amplified sensitivity has been developed for the detection of tissue polypeptide antigen (TPA). The nano-Au monolayer on glass chip was controllably prepared using 3-mercaptopropyltrimethoxysilane (MPTMOS) as linker by a solution self-assembly method. To analyze its quality, the nano-Au monolayer was characterized by UV-visible spectrum and atomic force microscopy (AFM). The resulting chip was modified by tissue polypeptide antigen antibody (anti-TPA), and then bovine serum albumin (BSA) was employed to block the possible remaining active sites of the nano-Au monolayer. An immunosensor was constructed with biocompatible monolayer nano-Au membrane and the desirable TPA antibody/BSA composite membrane, and it showed good selectivity, high sensitivity and a wide linear response to TPA in the range of 1-1000 ng L(-1) with a detection limit of 1.2 x 10(-3) ng L(-1), as well as good stability and long-term life. Owing to its ease of operation, low detection limit and low cost, it is expected that the proposed procedure may hold great promise in both research-based and clinical assays.

The changes of brainstem auditory evoked potentials (BAEP) after vertebrobasilar artery ischemia in rabbits.

Brainstem auditory evoked potentials (BAEPs) have been used as a valuable neurophysiologic index of neuronal dysfunction in the level of the brainstem. BAEPs are also useful in subdividing evoked potentials into normal, slight, or pronounced in patients with vertebrobasilar insufficiency. We investigated the changes of BAEP after vertebrobasilar artery ischemia in rabbits and its significance in clinical work. A brainstem ischemic model was made by unilateral extracranial occlusion of vertebral artery to monitor BAEPs at 0, 10, 20, 30, 40, 50, and 60 min after occlusion. We found that peak latencies (PL) of I, III, and most notably V were gradually extended. In addition, we observed a significant (P < 0.05) delay of interpeak latencies (IPL) of waves I­III, III­V, and I­V after occlusion. This delay became more significant in IPL I­V 60 min after occlusion. Our results also demonstrate that the amplitude of I and V had no obvious change (P < 0.05). In the rabbit with bilateral extracranial occlusion of vertebral artery, BAEP waveforms disappeared 10 min after occlusion. Our results showed that vertebrobasilar insufficiency caused brainstem ischemia, which induced BAEP abnormity. Taken together, our findings suggest that BAEP has important significance for the clinical diagnosis of vertebrobasilar insufficiency. Therefore, early detection of neuronal change after transient cerebral ischemia is important in initiating treatment within the therapeutic window.

[Study on the infrared fingerprint of Evodia rutaecarpa based on the methods of sequential analysis of dual-indexes].

OBJECTIVE: To establish a method to identify the different origins of herba Evodia rutaecarpa by IR and provide a new technique for their identification and quality evaluation. METHODS: The herba materials were extracted by chloroform and absolute alcohol, the powder and the extracts of Evodia rutaecarpa were mixed and pelleted with KBr. The slides were detected within 4000 - 400 cm(-1) by FIR spectrophotometry. The Difference of samples was studied. RESULTS: The infrared spectrums of Evodia rutaecarpa extracted by chloroform were obviously different. CONCLUSION: The method can be used to identify and appraise the different origins of herba Evodia rutaecarpa.