The cell viability staining on day 1 (A) and 14 (B) of the culture.

<p>Bar  = 100 µm.</p

Sequences of primers and probes used for Real-Time PCR. Sequences related to gene type X collagen and MMP13 are proprietary to Applied Biosystems Inc. and not disclosed.

<p>Sequences of primers and probes used for Real-Time PCR. Sequences related to gene type X collagen and MMP13 are proprietary to Applied Biosystems Inc. and not disclosed.</p

The evaluation of the hypertrophic calcification of the hydrogels.

<p>Calcium content of the hMSC-laden hydrogels normalized to wet weight (w.t.) and dry weight (d.w) (A); and Von Kossa staining of the histological sections of the hydrogels after 28 days of culture, bar  = 500 µm, *p<0.05 vs. MMP group (n = 4).</p

The gene expression of type II collagen, aggrecan, type X collagen and MMP13 in fold changes on day 7, 14 or 28 of the culture normalized to pre-differentiated cells.

<p>*p<0.05 vs. MMP group (n = 4).</p

The analysis of the cartilaginous matrix contents of the hMSC-seeded hydrogels.

<p>GAG and collagen content (normalized to the DNA content) of hMSC-laden HA hydrogels (A) and immunohistochemistry staining of the hydrogel constructs against chondroitin sulfate, type II and I collagen after 28 days of culture (B), Bar  = 50 µm, *p<0.05 vs. MMP group (n = 4).</p

Substrate Coupling Strength of Integrin-Binding Ligands Modulates Adhesion, Spreading, and Differentiation of Human Mesenchymal Stem Cells

Substrate stiffness has been shown to regulate the differentiation fate of human mesenchymal stem cells (hMSCs). hMSCs sense and respond to substrate rigidity by exerting traction forces upon the binding between integrins and integrin-specific ligands present on the substrate surface. However, in previous studies, integrin-specific ligands such as Arg–Gly–Asp (RGD) peptides are always grafted to the substrate by a permanent covalent bond. Whether the coupling strength of integrin-specific ligands on substrate will influence cell behaviors has not been explored. In this work, we have developed a facile platform to investigate the effects of varied coupling strength between the RGD peptide and the glass substrate on stem cell behaviors. Glass coverslips are decorated with positive charges by silanization using (3-aminopropyl) triethoxysilane (APTES) to immobilize negatively charged citrate-capped gold nanoparticles (cit-AuNPs) solely via electrostatic interactions. The monolayer of electrostatically immobilized cit-AuNPs is further conjugated with the thiolated RGD peptides through the sulfur–gold bond. The substrate coupling strength of the RGD peptides, which is dependent on the electrostatic interactions between the APTES-treated glass substrate and the cit-AuNPs, is simply tuned by changing the APTES dosage and, hence, the resultant positive charge density on the surface. A total of 0.5% and 12.5% of APTES are used to fabricate low-coupling-strength surfaces (namely, LCS0.5 and LCS12.5), whereas 25% and 50% of APTES are used to fabricate high-coupling-strength surfaces (namely, HCS25 and HCS50). Fluorescence microscopy shows that hMSCs spread well and form stable actin filamentous structure on HCS surfaces but not on LCS surfaces. Remarkably, hMSCs exhibit enhanced osteogenesis on HCS surfaces as revealed by the immunostaining results of multiple early osteogenic markers. These differential behaviors may be governed by Yes-associated protein (YAP), a mechanosensitive transcriptional regulator of stem cells. Our findings highlight the importance of the substrate coupling strength of integrin-binding ligands on regulating adhesion, spreading, and differentiation of hMSCs

Data_Sheet_1_Dietary vitamin E intake and risk of Parkinson's disease: a cross-sectional study.docx

ObjectiveCurrent evidence on the association between dietary vitamin E intake and the risk of Parkinson's disease (PD) is limited. The aim of the study was to explore the association of dietary vitamin E intake with PD in the United States among adults over 40 years.MethodsWe conducted a cross-sectional study with data collected from National Health and Nutrition Examination Survey (NHANES) from 2009 to 2018. A total of the sample of 13,340 participants were included. To identify the different characteristics of the participants, we utilized propensity score matching (PSM) to reduce the effects of selection bias and confounding variables. Weighted univariate and multivariable logistic regression were used to examine the association between dietary vitamin E intake and PD before and after matching. Then, restricted cubic spline (RCS) was used to visually describe the possible non-linear relationships. Finally, we employed the subgroup analysis to further investigate the relationship between dietary vitamin E intake and PD.ResultsAccording to the weighted univariate and multivariable logistic regression analysis, vitamin E intake was inversely associated with the risk of PD before and after matching. The results of RCS analysis revealed no non-linear inverse relationship between vitamin E intake and PD before and after matching. The subgroup analysis showed that age may influence the negative association between vitamin E and PD (P ConclusionAmong participants over 40 years of age, vitamin E intake was negatively associated with the risk of PD. Our data may support the supplementation of vitamin E to be used as an intervention strategy for the occurrence of PD.</p

Data_Sheet_1_Effect of acupuncture for non-motor symptoms in patients with Parkinson’s disease: A systematic review and meta-analysis.doc

BackgroundAlthough non-motor symptoms of Parkinson’s disease (PD) are serious, effective treatments are still lacking. Acupuncture may have clinical benefits for non-motor symptoms of PD patients, but high-quality evidence supporting this possibility is still limited. Hence, we conducted this meta-analysis to evaluate the effect of acupuncture treatment on non-motor symptoms in patients with PD.MethodsRandomized controlled trials (RCTs) of acupuncture treatment for PD were retrieved from the following electronic databases: Medline (OVID), Embase (OVID), Cochrane Library, Web of Science, China National Knowledge Infrastructure, Chinese BioMedical Literature Database, Chonqing VIP (CQVIP), and Wangfang database. Studies evaluating non-motor symptoms of PD were retrieved. Methodological quality was assessed using the Cochrane Handbook for Systematic Reviews of Interventions.ResultsA total of 27 RCTs were included, among which 8 outcomes related to non-motor symptoms were evaluated. The results showed that acupuncture combined with medication had benefits for PD-related insomnia relative to medication alone or sham acupuncture [standardized mean difference (SMD) = 0.517; 95% confidence interval (CI) = 0.242–0.793; p = 0.000], and acupuncture treatment had benefits at 8 weeks (SMD = 0.519; 95% CI = 0.181–0.857; p = 0.003). Regarding depression, acupuncture treatment was more effective (SMD = −0.353; 95% CI = −0.669 to −0.037; p = 0.029) within 2 months (SMD = −0.671; 95% CI = −1.332 to −0.011; p = 0.046). Regarding cognition, quality of life, and Unified Parkinson’s Disease Rating Scale (UPDRS) I and II scores, acupuncture treatment was effective [SMD = 0.878, 95% CI = 0.046–1.711, p = 0.039; SMD = −0.690, 95% CI = −1.226 to −0.155, p = 0.011; weighted mean difference (WMD) = −1.536, 95% CI = −2.201 to −0.871, p = 0.000; WMD = −2.071, 95% CI = −3.792 to −0.351, p = 0.018; respectively]. A significant difference was not found in terms of PD-related constipation. Only one study evaluated PD-related fatigue.ConclusionThe results of the analysis suggested that acupuncture treatment could ameliorate the symptoms of depression, quality of life, cognition, total mentation, behavior and mood, and activities of daily living in PD patients. Nevertheless, more prospective, well-designed RCTs with larger sample sizes are required to confirm our findings.</p

A Gold@Polydopamine Core–Shell Nanoprobe for Long-Term Intracellular Detection of MicroRNAs in Differentiating Stem Cells

The capability of monitoring the differentiation process in living stem cells is crucial to the understanding of stem cell biology and the practical application of stem-cell-based therapies, yet conventional methods for the analysis of biomarkers related to differentiation require a large number of cells as well as cell lysis. Such requirements lead to the unavoidable loss of cell sources and preclude real-time monitoring of cellular events. In this work, we report the detection of microRNAs (miRNAs) in living human mesenchymal stem cells (hMSCs) by using polydopamine-coated gold nanoparticles (Au@PDA NPs). The PDA shell facilitates the immobilization of fluorescently labeled hairpin DNA strands (hpDNAs) that can recognize specific miRNA targets. The gold core and PDA shell quench the fluorescence of the immobilized hpDNAs, and subsequent binding of the hpDNAs to the target miRNAs leads to their dissociation from Au@PDA NPs and the recovery of fluorescence signals. Remarkably, these Au@PDA–hpDNA nanoprobes can naturally enter stem cells, which are known for their poor transfection efficiency, without the aid of transfection agents. Upon cellular uptake of these nanoprobes, we observe intense and time-dependent fluorescence responses from two important osteogenic marker miRNAs, namely, miR-29b and miR-31, only in hMSCs undergoing osteogenic differentiation and living primary osteoblasts but not in undifferentiated hMSCs and 3T3 fibroblasts. Strikingly, our nanoprobes can afford long-term tracking of miRNAs (5 days) in the differentiating hMSCs without the need of continuously replenishing cell culture medium with fresh nanoprobes. Our results demonstrate the capability of our Au@PDA–hpDNA nanoprobes for monitoring the differentiation status of hMSCs (i.e., differentiating versus undifferentiated) via the detection of specific miRNAs in living stem cells. Our nanoprobes show great promise in the investigation of the long-term dynamics of stem cell differentiation, identification and isolation of specific cell types, and high-throughput drug screening