Key Residue in the Precursor Region of M Protein Contributes to the Neurovirulence and Neuroinvasiveness of the African Lineage of Zika Virus.

The Zika virus (ZIKV) represents an important global health threat due to its unusual association with congenital Zika syndrome. ZIKV strains are phylogenetically grouped into the African and Asian lineages. However, the viral determinants underlying the phenotypic differences between the lineages remain unknown. Here, multiple sequence alignment revealed a highly conserved residue at position 21 of the premembrane (prM) protein, which is glutamic acid and lysine in the Asian and African lineages, respectively. Using reverse genetics, we generated a recombinant virus carrying an E21K mutation based on the genomic backbone of the Asian lineage strain FSS13025 (termed E21K). The E21K mutation significantly increased viral replication in multiple neural cell lines with a higher ratio of M to prM production. Animal studies showed E21K exhibited increased neurovirulence in suckling mice, leading to more severe defects in mouse brains by causing more neural cell death and destruction of hippocampus integrity. Moreover, the E21K substitution enhanced neuroinvasiveness in interferon alpha/beta (IFN-α/ß) receptor knockout mice, as indicated by the increased mortality, and enhanced replication in mouse brains. The global transcriptional analysis showed E21K infection profoundly altered neuron development networks and induced stronger antiviral immune response than wild type (WT) in both neural cells and mouse brains. More importantly, the reverse K21E mutation based on the genomic backbone of the African strain MR766 caused less mouse neurovirulence. Overall, our findings support the 21st residue of prM functions as a determinant for neurovirulence and neuroinvasiveness of the African lineage of ZIKV. IMPORTANCE The suspected link of Zika virus (ZIKV) to birth defects led the World Health Organization to declare ZIKV a Public Health Emergency of International Concern. ZIKV has been identified to have two dominant phylogenetic lineages, African and Asian. Significant differences exist between the two lineages in terms of neurovirulence and neuroinvasiveness in mice. However, the viral determinants underlying the phenotypic differences are still unknown. Here, combining reverse genetics, animal studies, and global transcriptional analysis, we provide evidence that a single E21K mutation of prM confers to the Asian lineage strain FSS130125 significantly enhanced replication in neural cell lines and more neurovirulent and neuroinvasiveness phenotypes in mice. Our findings support that the highly conserved residue at position 21 of prM functions as a determinant of neurovirulence and neuroinvasiveness of the African lineage of ZIKV in mice.

Asymmetric Transfer Hydrogenation of Ketones Improved by PNN-Manganese Complexes.

Chiral manganese(I) complexes that contain carbocyclic-fused 8-amino-5,6,7,8-tetrahydroquinolinyl groups that are appended with distinct para-R substituents have proven to be effective catalysts in the asymmetric transfer hydrogenation (ATH) of a wide range of ketones (48 examples). Notably, Mn2 proved to be the most productive catalyst, allowing an outstanding turnover number of 8300 with catalyst loadings as low as 0.01 mol %. Furthermore, this catalytic protocol shows considerable promise for applications in the synthesis of chiral drugs such as Lusutrombopag.

Construction of controlled hyper-crosslinked nanofibrous tubes for Cr(VI) removal: Response surface, kinetics, and isotherm.

Porous organic polymers (POPs) exhibit significant potential for adsorbing toxic metal ions in wastewater. Developing POPs with controlled morphologies is a pivotal direction in this field. This study synthesized a series of novel hyper-crosslinked nanofibrous tubes designated HCNT-Cn (n = 4, 8, 12, 16) via Friedel-Crafts alkylation and quaternization reactions. These reactions were fine-tuned through a post-synthetic strategy involving varying alkyl chain lengths. These materials were characterized using FT-IR, SEM, N2 adsorption-desorption isotherms, among others, and they were specifically evaluated for their ability to adsorb Cr(VI). Among the variants, HCNT-C4 exhibited the highest specific surface area (495.26 m2 g-1), superior hydrophilicity (CA = 48.7°), and optimal adsorption performance. The adsorption kinetics of HCNT-C4 conformed to a pseudo-second-order model, while its adsorption isotherm aligned with the Langmuir model. An investigation into the impact of Cr(VI) removal was conducted using three independent variables in a Central Composite Design (CCD) response surface model, revealing that under optimal conditions, the Cr(VI) removal efficiency reached 98%. Additionally, a mechanism for Cr(VI) adsorption on HCNT-C4 was proposed. It was also found that HCNT-C4 could be reused up to four times, maintaining a removal efficiency of 70%. This study suggests potential applications for removing Cr(VI) from contaminated wastewater.

Association between abnormal body weight and stroke outcome: A meta-analysis and systematic review.

BACKGROUND AND PURPOSE: To test the hypothesis that "obesity paradox" exists in stroke patients, we conducted a meta-analysis and systematic review on the association between abnormal body weight (obesity, overweight, or underweight) and the outcome of different types of stroke. METHODS: This meta-analysis and systematic review was performed in conformity to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines in Appendix S2. Studies investigating the association between abnormal body weight and the outcome of different types of stroke were searched for in the PubMed and Embase databases from their inception to 20 March 2021. RESULTS: Thirty-three articles including 84,660 patients were included in this study. Obesity and overweight were associated with longer survival in mixed-stroke patients (acute ischemic stroke [AIS] combined with one or more other stroke subtypes) than was normal weight, whereas underweight was related to shorter survival; the pooled hazard ratios (HRs) of mortality were 0.77 (95% confidence interval [CI] = 0.71-0.83) for obesity, 0.76 (95% CI = 0.72-0.80) for overweight, and 1.71 (95% CI = 1.56-1.87) for underweight. However, only obesity was associated with longer survival in AIS patients compared with normal weight, and underweight was related to shorter survival; the pooled HR of mortality was 0.75 (95% CI = 0.64-0.88) for obesity and 1.53 (95% CI = 1.27-1.85) for underweight. After merging mixed-stroke and AIS patients, we obtained similar results as in mixed-stroke patients. CONCLUSIONS: Our results suggested that in patients with mixed stroke or AIS, obesity was associated with a longer survival time than normal weight, whereas underweight was associated with a shorter survival time.

Safety and efficacy of tirofiban combined with endovascular therapy compared with endovascular therapy alone in acute ischemic stroke: a meta-analysis.

Endovascular treatment (EVT) has been widely used for treating acute ischemic stroke (AIS). However, the safety and efficacy of treating AIS with tirofiban combined with EVT remain controversial. Therefore, we conducted a meta-analysis to evaluate this treatment. Randomized controlled trials and cohort studies that compared treatment with tirofiban combined with EVT and EVT alone were included in our meta-analysis. Those published from inception to March 31, 2020, were searched using the PubMed, Web of Science, Embase, and Cochrane Library databases. Safety was assessed based on symptomatic intracranial hemorrhage (sICH) incidence and 3-month mortality. Efficacy was assessed based on modified Rankin Scale (mRS) scores at 3 months post-EVT and recanalization rates. Data were analyzed using either the random-effects or fixed-effects model based on the heterogeneity of studies. In total, one RCT, six prospective studies, and four retrospective studies (2387 AIS cases) were assessed. Our meta-analysis showed that tirofiban combined with EVT did not increase sICH risk (RR, 1.06; 95%CI, 0.79 to 1.42; P = 0.72) and 3-month mortality (RR, 0.87; 95%CI, 0.74 to 1.04; P = 0.12). Recanalization rates were not significantly different between patients treated with tirofiban combined with EVT and those treated with EVT alone (RR, 1.04; 95%CI, 1.00 to 1.08; P = 0.07), but tirofiban combined with EVT was significantly associated with favorable functional outcomes (mRS score, 0-2) in AIS patients (RR, 1.13; 95%CI, 1.02 to 1.25; P = 0.02). Tirofiban combined with EVT appears to be safe and potentially effective in treating AIS.

The Impact of Remote Ischaemic Conditioning on Beat-to-Beat Heart Rate Variability Circadian Rhythm in Healthy Adults.

BACKGROUND: Remote ischaemic conditioning (RIC) is an intervention that may exert a protective effect over multiple tissues or organs by regulating neuronal signal transduction. Heart rate variability (HRV) can assess the state of the autonomic nervous system. However, whether RIC can also regulate HRV in humans remains unknown. METHOD: This was a self-controlled interventional study in which serial beat-to-beat monitoring was performed at the same seven time points (7, 9, and 11 AM; 2, 5, and 8 PM; and 8 AM on the next day) with or without RIC in 50 healthy adults. The seven time points on the RIC day were defined as baseline, 1 hour, 3 hours, 6 hours, 9 hours, 12 hours, and 24 hours after RIC. The RIC protocol consisted of 4×5-minute inflation/deflation in one arm and one thigh cuff at 200 mmHg pressure from 7:20 to 8 AM. This study is registered on ClinicalTrials.gov (NCT02965547). RESULTS: We included 50 healthy adult volunteers (aged 34.54±12.01 years, 22 men [44%], all Asian). The variables analysed in frequency-domain measures performed as power of low-frequency in normalised units (0.04-0.15 Hz), high-frequency in normalised units (0.15-0.40 Hz), and ratio of low frequency to high frequency. The time-domain parameters standard deviation (SD) of all normal to normal (NN) intervals (SDNN), mean of the 5-minute SD of the NN intervals, SD of the consecutive 5-minute averages of NN intervals, and the root mean square of successive differences of NN intervals, and time-domain parameters calculated from Poincaré plots, SD of the short diagonal axis in Poincaré plot (SD1), SD of the long diagonal axis in Poincaré plot (SD2), and SD1/SD2 were also obtained. The SDNN and SD2 significantly increased 1 hour after RIC (p=0.029 and p=0.045, respectively). Additionally, the SD2 increased a second time 12 hours after RIC (p=0.041), which represented inhibited sympathetic activity. CONCLUSIONS: Heart rate variability increase and sympathetic inhibition induced by RIC appeared both on the early and delayed protective window of RIC, which may indicate some of the underlying mechanisms by which RIC may offer protection.

Mapping a double flower phenotype-associated gene DcAP2L in Dianthus chinensis.

The double flower is a highly important breeding trait that affects the ornamental value in many flowering plants. To get a better understanding of the genetic mechanism of double flower formation in Dianthus chinensis, we have constructed a high-density genetic map using 140 F2 progenies derived from a cross between a single flower genotype and a double flower genotype. The linkage map was constructed using double-digest restriction site-associated DNA sequencing (ddRAD-seq) with 2353 single nucleotide polymorphisms (SNPs). Quantitative trait locus (QTL) mapping analysis was conducted for 12 horticultural traits, and major QTLs were identified for nine of the 12 traits. Among them, two major QTLs accounted for 20.7% and 78.1% of the total petal number variation, respectively. Bulked segregant RNA-seq (BSR-seq) was performed to search accurately for candidate genes associated with the double flower trait. Integrative analysis of QTL mapping and BSR-seq analysis using the reference genome of Dianthus caryophyllus suggested that an SNP mutation in the miR172 cleavage site of the A-class flower organ identity gene APETALA2 (DcAP2L) is responsible for double flower formation in Dianthus through regulating the expression of DcAG genes.

Venous sinus stenting improves cerebral autoregulation in a patient with venous sinus stenosis: a case report.

BACKGROUND: Venous sinus stenosis (VSS) is a type of cerebral venous vascular disease. Cerebral autoregulation is an indicator of cerebral arterial function. The cerebral circulatory system is composed of the venous system and arterial system. Impaired venous function may affect arterial function. Thus, cerebral venous stenosis may influence cerebral autoregulation. CASE PRESENTATION: In this case, a 50-year-old woman with transient blindness and headache was admitted to the hospital. The patient was diagnosed with VSS. A stent was placed at the stenosis. The stent released the intravenous pressure and remitted the patient's symptoms. Measurements of dynamic cerebral autoregulation (dCA) were performed at 3 time points: before stenting, after stenting, and 3 months later. The dCA gradually improved after stenting. CONCLUSION: VSS may have an influence on cerebral autoregulation, and effective treatment improves cerebral autoregulation in patients with VSS.

Hydrogenation of CO2 to Methanol Catalyzed by Cp*Co Complexes: Mechanistic Insights and Ligand Design.

A direct hydride transfer mechanism with three cascade cycles for the conversion of carbon dioxide and dihydrogen to methanol (CO2 + 3H2 → CH3OH + H2O) catalyzed by a half-sandwich cobalt complex [Cp*Co(bpy-Me)OH2]2+ (1) is proposed based on density functional theory calculations. The formation of methanediol via hydride transfer from Co to formic acid (4 → TS8,11) is the rate-determining step with a total barrier of 26.0 kcal/mol in free energy. Furthermore, 15 analogues of 1 are constructed by replacing the hydrogen atoms at the two meta and para positions of the bipyridine ligand with different functional groups (1b-1l), the carbon atoms in the bipyridine ligand with nitrogen atoms (1m-1o), and one pyridine ligand with N-heterocyclic carbene (1p). Among all newly proposed complexes, [Cp*Co(2,2'-bipyrazine)OH2]2+ (1n) is the most active one with a total barrier of 19.6 kcal/mol in free energy. Such a low barrier indicates 1n is a promising catalyst for efficient conversion of CO2 and H2 to methanol at room temperature.

Characteristics of cardio-Cerebrovascular modulation in patients with generalized anxiety disorder: an observational study.

BACKGROUND: Generalized anxiety disorder (GAD) has been shown in previous studies to display abnormal cerebral blood flow velocity (CBFV); however, the characteristics of cardio-cerebrovascular modulation are unknown. We aimed to analyze cardio-cerebrovascular modulation using parameters from a supine-to-standing test. METHODS: There are 2 parts to this study; in Part 1, 125 participants with Hamilton Anxiety scale scores ≥14 were enrolled, and 33 age- and sex-matched medically and psychiatrically healthy volunteers were recruited as control participants. Patients were divided by score into mild, moderate, and severe anxiety groups. The cardio-cerebrovascular modulation using the parameters of dynamic changes of CBFV and heart rate in response to an orthostatic challenge were investigated. In Part 2, we followed up the severe GAD patients for 6 months and repeated the supine-to-standing test, and severe GAD patients were divided into recovery and non-recovery groups. RESULTS: In part 1, the GAD group displayed more marked CBFV and heart rate changes than the healthy group, but there was no difference in the CBFV and heart rate changes from the supine to upright position in mild, moderate, and severe anxiety groups. In part 2, The recovery group demonstrated significant improvement in changes in the CBFV and heart rate values from the supine to the upright position after treatment compared with before treatment. In the non-recovery group, the CBFV and heart rate changes were significantly higher than the healthy group regardless of treatment. CONCLUSIONS: Cardio-cerebrovascular modulation is compromised in patients with GAD, however, this impairment can be restored to normal after the disappearance of anxiety.

[Genetic analysis of two pediatric patients with Beckwith-Wiedemann syndrome].

OBJECTIVE: To explore the genetic cause for two children with omphalocele. METHODS: The patients were examined, and the medical history of their families was collected. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was performed to detect potential mutation in the patients. RESULTS: Loss of methylation of imprinting center 2 (IC2) at the 11p15.5 region of the maternal chromosome was detected in both children. CONCLUSION: The two patients were diagnosed with Beckwith-Wiedemann syndrome by MS-MLPA. The loss of methylation of IC2 probably underlies the disease in both patients.

Directivity pattern of the sound radiated from axisymmetric stepped plates.

For the purpose of optimal design and efficient utilization of the kind of stepped plate radiator in air, in this contribution, an approach for calculation of the directivity pattern of the sound radiated from a stepped plate in flexural vibration with a free edge is developed based on Kirchhoff-Love hypothesis and Rayleigh integral principle. Experimental tests of directivity pattern for a fabricated flat plate and two fabricated plates with one and two step radiators were carried out. It shows that the configuration of the measured directivity patterns by the proposed analytic approach is similar to those of the calculated approach. Comparison of the agreement between the calculated directivity pattern of a stepped plate and its corresponding theoretical piston show that the former radiator is equivalent to the latter, and the diffraction field generated by the unbaffled upper surface may be small. It also shows that the directivity pattern of a stepped radiator is independent of the metallic material but dependent on the thickness of base plate and resonant frequency. The thicker the thickness of base plate, the more directive the radiation is. The proposed analytic approach in this work may be adopted for any other plates with multi-steps.

Morphological manifestation of tuberculous pleurisy in children under medical thoracoscope and diagnostic value.

OBJECTIVE: Our study analyzed the main manifestations of tuberculous pleurisy (TBP) in children under medical thoracoscopy (MT). This article aimed to explore the clinical application value of MT in the diagnosis and treatment of TBP in children. METHODS: In our study, we selected 23 TBP patients diagnosed in our hospital. We analyzed the clinical data and thoracoscopic morphology of these patients. At the same time, we also observed the pathological manifestations, acid-fast staining, and treatment effects of the patient's diseased tissue under MT. RESULTS: The MT clinical findings of TBP patients include pleural hyperemia and edema, miliary nodules, scattered or more white nodules, simple pleural adhesion, wrapped pleural effusion, massive cellulose exudation, yellow-white caseous necrosis, pleural hyperplasia and hyperplasia, and mixed pleural necrosis. The positive rate of pleural biopsy was 73.91% and that of acid-fast staining was 34.78%. The main pathologic types of these patients were tuberculous granulomatous lesions (16 cases), caseous necrosis (5 cases), and fibrinous exudative, multinucleated giant cell and other inflammatory cell infiltration lesions (13 cases). The average time of diagnosis of the 23 patients was 8.32 days (5.0-16.0 days), and they were transferred to specialized hospitals for treatment after diagnosis. The mean time of chest drainage was 3.0-5.0 days after treatment. The average time for their body temperature to return to normal was 3.31 days (2.0-5.0 days). CONCLUSION: Thoracoscopic lesions of TBP in children are varied. The use of MT is not only helpful for the early diagnosis and treatment of TBP. It also protects and improves lung function. Therefore, the use of MT has high clinical value.

Optimal cut-off values of visceral fat area for predicting metabolic syndrome among patients with ischemic stroke: a cross-sectional study.

Objectives: The prevalence of metabolic syndrome (MetS) among patients with ischemic stroke is relatively high. The visceral fat area (VFA) is a predictor of MetS. This study aimed to estimate sex-specific optimal cut-off values of VFA and MetS risk factors among patients with ischemic stroke. Methods: A cross-sectional study including 851 patients with ischemic stroke was conducted between March 2019 and January 2020 in a tertiary hospital in the northeast of China. VFA was measured using the dual bioelectrical impedance method. Binary logistic regression analysis was used to investigate MetS risk factors, and the VFA cut-off value was assessed using receiver operating characteristic curve analysis. Results: The overall prevalence of MetS was 43.4%. After adjusting for potential confounders, female sex (odds ratio [OR] = 2.86, p < 0.001), the presence of visceral obesity according to VFA (OR = 7.45, p < 0.001), being overweight (OR = 2.75, p < 0.001) or obesity (OR = 6.00, p < 0.001) were associated with an increased risk of MetS. The correlation between VFA and MetS in patients with ischemic stroke was strongest with cut-off values of 104.3 cm2 (sensitivity 73.0%, specificity 83.1%) for men, and 94.1 cm2 (sensitivity 70.9%, specificity 72.9%) for women. Conclusion: MetS affected approximately a half of patients with ischemic stroke. Female sex, visceral obesity, and body mass index were independent risk factors for the development of MetS. Sex-specific reference values for VFA are proposed for the prediction of incident MetS in patients with ischemic stroke.

Effects of a Planned Web-Based Educational Intervention Based on the Health Belief Model for Patients With Ischemic Stroke in Promoting Secondary Prevention During the COVID-19 Lockdown in China: Quasi-Experimental Study.

Background: Some common modified vascular risk factors remain poorly controlled among stroke survivors, and educational programs may help improve these conditions. Objective: This study aimed to evaluate the effect of a planned web-based educational intervention based on the health belief model (HBM) in promoting secondary prevention among patients with ischemic stroke. Methods: An evaluation-blinded quasi-experimental trial with a historical control group was conducted. Patients admitted from March to June 2020 were assigned to the historical control group, and patients admitted from July to October 2020 were assigned to the intervention group. The control group received routine health management. The intervention group received 6 additional sessions based on the HBM via Tencent Meeting, an audio and video conferencing application, within 3 months after discharge. Sessions were held every 2 weeks, with each session lasting approximately 40 minutes. These sessions were conducted in small groups, with about 8 to 10 people in each group. The primary outcomes were changes in blood pressure (BP), low-density lipoprotein cholesterol (LDL-C), hemoglobin A1c (HbA1c), and the proportion of patients achieving the treatment target. The secondary outcomes were medication adherence, assessed with the Morisky Medicine Adherence Scale (MMAS), and disability, assessed with the modified Rankin scale. Results: In total, 315 patients experiencing their first-ever stroke were analyzed. More patients in the intervention group had controlled BP (41.9% vs 28.4%; adjusted odds ratio [aOR] 1.93; P=.01), LDL-C (83.1% vs 67.7%; aOR 2.66; P=.001), and HbA1c (91.9% vs 83.9%; aOR: 3.37; P=.04) levels as well as a significant postintervention decrease in the systolic BP (adjusted ß -3.94; P=.02), LDL-C (adjusted ß -0.21; P=.008), and HbA1c (adjusted ß -0.27; P<.001), compared with control groups. Significant between-group differences were observed in medication adherence (79.4% vs 63.2%; aOR 2.31; P=.002) but not in favorable functional outcomes. Conclusions: A web-based education program based on the HBM may be more effective than current methods used to educate patients having strokes on optimal vascular risk factors and medication adherence.

Identification of mutations in viral proteins involved in cell adaptation using a reverse genetic system of the live attenuated hepatitis A virus vaccine H2 strain.

The live attenuated hepatitis A virus vaccine H2 strain was developed by passaging a wild-type H2w isolate in cell cultures. Currently, the mechanism underlying its attenuation phenotype remain largely unknown. In this study, we generated a full-length infectious cDNA clone of the H2 strain using in-fusion techniques. The recovered H2 strain (H2ic) from the cDNA clone exhibited an efficient replication in both the hepatoma cell line Huh7.5.1 and the 2BS cell line used for vaccine production, similar to the parental H2 strain. Additionally, H2ic did not cause disease in Ifnar1-/- C57 mice, consistent with the H2 strain. To explore the cell-adaptive mutations of the H2 strain, chimeric viruses were generated by replacing its non-structural proteins with corresponding regions from H2w using the infectious cDNA clone as a genetic backbone. The chimeric viruses carrying the 3C or 3D proteins from H2w showed decreased replication in Huh7.5.1 and 2BS cell lines compared to H2ic. Other chimeric viruses containing the 2B, 2C, or 3A proteins from H2w failed to be recovered. Furthermore, there were no significant differences in disease manifestation in mice between H2ic and the recovered chimeric viruses. These results demonstrate that adaptive mutations in the 2B, 2C, and 3A proteins are essential for efficient replication of the H2 strain in cell cultures. Mutations in the 3C and 3D proteins contribute to enhanced replication in cell cultures but did not influence the attenuated phenotypes in mice. Together, this study presents the first reverse genetic system of the H2 strain and identifies viral proteins essential for adaptation to cell cultures.

Labile organic matter favors a low N2O yield during nitrogen removal in estuarine sediments.

Estuary harbors the active sediment denitrification and nitrous oxide (N2O) emission, while the knowledge of environmental controls on the denitrification-derived N2O yield remains underexplored. Here, we quantitatively assess the potential and in situ rates of N2O production during sediment denitrification in the Pearl River Estuary (PRE), China. Organic matter determines the product stoichiometry and capacity of nitrogen removal. In particular, labile organic matter (LOM) reduces N2O yield via enhancing the complete coupled nitrification-denitrification. Our results reveal that the chain processes, primary production-LOM settling-sedimentary respiration-coupled nitrification-denitrification, control the sediment denitrification and N2O production, linking the carbon and nitrogen biogeochemical cycles in the atmosphere-water column-sediment continuum. The PRE sediments serve as nitrogen removal hotspots but with low efficiency (~25 % of riverine input) and strong N2O release (~66 % of daily sea-air N2O efflux). These findings contribute to policy makers to develop knowledge-based management actions for achieving sustainable coastal environments and mitigating N2O emission.

Effect of acupuncture for patients with knee osteoarthritis: study protocol for a double-dummy randomized controlled trial.

BACKGROUND: Acupuncture has been used to relieve chronic pain in patients with knee osteoarthritis (KOA), but the evidence is contradictory. Therefore, we carefully designed a double-dummy randomized controlled trial (RCT) to explore the therapeutic effect of acupuncture for KOA. METHODS/DESIGN: A total of 138 eligible participants with KOA who consent to participate will be randomly divided into Groups A, B, and C in a ratio of 1:1:1. Participants in Group A will receive verum acupuncture and placebo gel, while those in Groups B and C will be treated with diclofenac diethylammon gel and sham acupuncture, sham acupuncture and placebo gel, respectively. The patients will receive 4 weeks of treatment, five times a week, including acupuncture treatment once a day for 30 min and gel treatment three times a day. The primary outcome will be the change of Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) at week 4. The secondary outcomes will include visual analog scale (VAS), Arthritis Quality of Life Measurement Scale Simplified Scale (AIMS2-SF), Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI) and Credibility/Expectancy Questionnaire. The evaluation will be performed at baseline, week 4, 8, and 12 after randomization. DISCUSSION: This double-dummy RCT used diclofenac diethylammon gel as a positive control, and the completion of this trial will provide detailed and accurate evidence of the efficacy and safety of acupuncture for KOA. TRIAL REGISTRATION: China Clinical Trials Registry No.ChiCTR2100043947. Registered on September 24, 2020. https://www.chictr.org.cn/showproj.html?proj=122536 .

Huangqi Decoction, a compound Chinese herbal medicine, inhibits the proliferation and activation of hepatic stellate cells by regulating the long noncoding RNA-C18orf26-1/microRNA-663a/transforming growth factor-ß axis.

OBJECTIVE: Huangqi Decoction (HQD), a classical traditional Chinese medicine formula, has been used as a valid treatment for alleviating liver fibrosis; however, the underlying molecular mechanism is still unknown. Although our previous studies showed that microRNA-663a (miR-663a) suppresses the proliferation and activation of hepatic stellate cells (HSCs) and the transforming growth factor-ß/small mothers against decapentaplegic (TGF-ß/Smad) pathway, whether long noncoding RNAs (lncRNAs) are involved in HSC activation via the miR-663a/TGF-ß/Smad signaling pathway has not yet reported. The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis. METHODS: The expression levels of lnc-C18orf26-1, miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction. HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs. The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs. Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs. RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs. Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA. The expression levels of collagen α-2(I) chain (COL1A2), α-smooth muscle actin (α-SMA) and TGF-ß/Smad signaling pathway-related proteins were determined using Western blotting. RESULTS: Lnc-C18orf26-1 was upregulated in TGF-ß1-activated HSCs and competitively bound to miR-663a. Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation, downregulated TGF-ß1-stimulated α-SMA and COL1A2 expression, and inhibited the TGF-ß1/Smad signaling pathway. HQD suppressed the proliferation and activation of HSCs. HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs. Further studies showed that HQD inhibited the expression of COL1A2, α-SMA, TGF-ß1, TGF-ß type I receptor (TGF-ßRI) and phosphorylated Smad2 (p-Smad2) in HSCs, and these effects were reversed by miR-663a inhibitor treatment. CONCLUSION: Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis. HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-ß1/TGF-ßRI/p-Smad2 axis.

Development and Optimization of Network Music Course Resources Based on Data Mining Technology under the Personalized Online Education Environment.

The ever-evolving Internet technology has brought more information resources for music teaching than ever before. For music educators, figuring out how to create and use these tools more effectively has become a pressing concern. China will usher in a new age of innovation and integration between the Internet and different traditional businesses with the idea of "Internet Plus." MOOCs present significant challenges to traditional teachers and teaching methods as a typical practice of the transformation of education informatization to innovation, and they also drive global education toward a new process of informatization development. This paper adopts Internet thinking as the guiding ideology for the development of future music education concepts and, concurrently, conducts specific transformation research based on Internet technology from four aspects: music teaching content, music teaching methods, music education concepts, and the passing down of traditional music culture. The improved association rule algorithm is used to organise the students' access data, decide which high-frequency websites are accessible to the students, and realize the intuitive positioning of high-frequency access and learning issues. Assist the school's decision-making and educational levels in identifying the issues that students have with their learning, and then, improve the network teaching platform's material to complete the task of personalized network education.