Table_1_The effect of basic medical insurance on the changes of primary care seeking behavior: An application of hierarchical age-period-cohort analysis.docx

In order to encourage residents to go to primary care facilities, China has set up differentiated basic medical insurance reimbursement ratios. The study aims to use the dynamic point of view of longitudinal data to examine the changes in the impact of basic medical insurance on primary care. The data for this study comes from the Chinese Family Panel Study (CFPS) in 2010, 2012, 2014, 2016, and 2018. We adopted Hierarchal Age-period-cohort-Cross-Classified Random Effects Models (HAPC-CCREM) to examine the changes in the impact of basic medical insurance on primary care. Compared with non-insured groups, participants of the New Rural Cooperative Medical System (coefficient = 0.730) have a relatively high incidence of primary care seeks, while Urban Residents' Basic Medical Insurance (coefficient = −0.482) and Urban Employees' Basic Medical Insurance (coefficient = −0.663) are lower, respectively. Age, period over time and cohort have a more obvious moderating effect on primary care seeks. The study of primary care behavior is an important direction for the construction of a hierarchical medical system. As basic medical insurance is the source of power for the hierarchical medical system, we can provide certain direction for policy formulation on the changes of basic medical insurance in primary care behavior.</p

Data_Sheet_1_Age and cohort trends of the impact of socioeconomic status on dietary diversity among Chinese older adults from the perspective of urban–rural differences: A prospective cohort study based on CLHLS 2002–2018.docx

The association between socioeconomic status (SES) and dietary diversity score (DDS) has been widely discussed, but little is known about the age and cohort effects on DDS and how the SES effect on DDS varies with age and across successive cohorts among urban and rural older adults in China. Thus, this study aimed to examine the temporal change in DDS among Chinese older adults and SES heterogeneities in such change from the perspective of urban–rural differentiation. Data from the Chinese Longitudinal Healthy Longevity Survey (CLHLS) between 2002 and 2018 were used, and a total of 13,728 participants aged between 65 and 105 years were included in this study. A total of eight food groups were used to assess DDS, while education, family income, and perceived income status were used to assess SES. A linear mixed model was used to estimate the age and cohort effects on DDS and their urban–rural and SES disparities. The results show that higher SES, including more education, family income, and perceived income status, was associated with higher DDS (for urban older adults, β = 0.1645, p = 0.0003, β = 0.2638, p < 0.0001, β = 0.2917, p < 0.0001, respectively; for rural older adults, β = 0.0932, p = 0.0080, β = 0.4063, p < 0.0001, β = 0.2921, p < 0.0001, respectively). The DDS of older adults increased with age and across successive cohorts in both urban and rural China. Moreover, we found the three-way interaction effect of SES, age, and cohort was statistically significant in both urban and rural China. Thus, living in an urban area and having higher SES are associated with higher DDS, but these associations change with age and across successive cohorts. The dietary health of earlier cohorts and rural oldest-old in China deserves more attention.</p

MicroRNA-181 Regulates CARM1 and Histone Aginine Methylation to Promote Differentiation of Human Embryonic Stem Cells

<div><p>As a novel epigenetic mechanism, histone H3 methylation at R17 and R26, which is mainly catalyzed by coactivator-associated protein arginine methyltransferase 1 (CARM1), has been reported to modulate the transcription of key pluripotency factors and to regulate pluripotency in mouse embryos and mouse embryonic stem cells (mESCs) in previous studies. However, the role of CARM1 in human embryonic stem cells (hESCs) and the regulatory mechanism that controls <em>CARM1</em> expression during ESCs differentiation are presently unknown. Here, we demonstrate that CARM1 plays an active role in the resistance to differentiation in hESCs by regulating pluripotency genes in response to BMP4. In a functional screen, we identified the miR-181 family as a regulator of <em>CARM1</em> that is induced during ESC differentiation and show that endogenous miR-181c represses the expression of <em>CARM1</em>. Depletion of CARM1 or enforced expression of miR-181c inhibits the expression of pluripotency genes and induces differentiation independent of BMP4, whereas overexpression of <em>CARM1</em> or miR-181c inhibitor elevates Nanog and impedes differentiation. Furthermore, expression of <em>CARM1</em> rescue constructs inhibits the effect of miR-181c overexpression in promoting differentiation. Taken together, our findings demonstrate the importance of a miR-181c-CARM1 pathway in regulating the differentiation of hESCs.</p> </div

DataSheet_1_Longitudinal evaluation of innate immune responses to three doses of CoronaVac vaccine.docx

The adaptive immune responses induced by inactivated COVID-19 vaccine has been extensively studied. However, few studies have analyzed the impact of COVID-19 vaccination on innate immune cells. Here in this study, we recruited 62 healthcare workers who received three doses of CoronaVac vaccine and longitudinally profiled the alterations of peripheral monocytes and NK cells during vaccination. The results showed that both the monocyte and NK cell subsets distribution were altered, although the frequencies of the total monocyte and NK cells remained stable during the vaccination. Additionally, we found that both the 2nd and 3rd dose of CoronaVac vaccination elicited robust IFN-γ-producing NK cell response. Our data provided necessary insights on innate immune responses in the context of three homologous CoronaVac dose vaccination, and supplied immunological basis for the future design of inactivated vaccines against SARS-CoV-2 or other viruses.</p

Supplementary document for Highly sensitive and label-free detection of biotin using liquid crystal-based optofluidic biosensor - 6416490.pdf

Supporting Informatio

DataSheet_2_Longitudinal evaluation of innate immune responses to three doses of CoronaVac vaccine.docx

The adaptive immune responses induced by inactivated COVID-19 vaccine has been extensively studied. However, few studies have analyzed the impact of COVID-19 vaccination on innate immune cells. Here in this study, we recruited 62 healthcare workers who received three doses of CoronaVac vaccine and longitudinally profiled the alterations of peripheral monocytes and NK cells during vaccination. The results showed that both the monocyte and NK cell subsets distribution were altered, although the frequencies of the total monocyte and NK cells remained stable during the vaccination. Additionally, we found that both the 2nd and 3rd dose of CoronaVac vaccination elicited robust IFN-γ-producing NK cell response. Our data provided necessary insights on innate immune responses in the context of three homologous CoronaVac dose vaccination, and supplied immunological basis for the future design of inactivated vaccines against SARS-CoV-2 or other viruses.</p

The miR-181 family directly regulates <i>CARM1</i> expression in hESC.

<p>(A) The expression levels of mature miRNAs predicted to target the <i>CARM1</i> 3′UTR were monitored in differentiated ESCs by qRT-PCR and normalized to endogenous <i>U6</i> expression. *, p<0.05; **, p<0.01. (B) The chromosome positions of the primary transcripts of miR-181 family members are shown, their expression levels in differentiated ESCs were determined by qRT-PCR and normalized to endogenous <i>β-actin</i> expression. **, p<0.01. (C) The predicted consequential pairing of miRNAs and their target regions in the wild-type <i>CARM1</i> 3′UTR or the mutant (mut) <i>CARM1</i> 3′UTR are shown. (D) The HEK293 cells were co-transfected with the firefly-luciferase-expressing vector pMIR-REPORT containing the wild-type <i>CARM1</i> 3′UTR, the mut <i>CARM1</i> 3′UTR or the control insert as well as the internal control renilla-luciferase-expressing vector pRL-TK and the indicated RNAs. After 48 h, the luciferase activities were measured. The data were normalized by dividing firefly luciferase activity with that of Renilla luciferase, **, p<0.01. (D) Luciferase activities were measured in human ESCs co-transfected with the pMIR-REPORT plasmid containing wild-type <i>CARM1</i> 3′UTR, mut <i>CARM1</i> 3′UTR or control insert and the internal control, pRL-TK. All the samples were assayed in duplicate (n = 3). **, p<0.01.</p

The miR-181c inhibitor suppresses hESC differentiation.

<p>(A) Enforced expression of the miR-181c inhibitor down-regulated mature miR-181c levels relative to negative control (NC) RNA-transfected ESCs, as shown by qRT-PCR. **, p<0.01. Western blotting detected that CARM1 and H3R17me2 protein levels were clearly decreased. (B, C, D) The effect of miR-181c inhibition on differentiated hESCs. Human ESCs were transfected with miR-181c inhibitor or NC RNA and then induced to differentiate by the addition of BMP4 in the absence of bFGF. (B) The expression of <i>CARM1</i>, <i>Nanog</i>, <i>Sox2</i>, and <i>Oct4</i> at the mRNA level was quantified by qRT-PCR, CARM1 and Nanog protein expression levels were also quantified by Western blotting. (C) Pluripotency was examined by AP staining 3 days after transfection. The counts of AP-positive clones and the images of the representative plates are shown. Samples were assayed in duplicate (n = 3). **, p<0.01. (D) Expression of a subset of differentiation-associated genes in ESCs transfected with miR-181c inhibitor or NC RNA were monitored by qRT-PCR and normalized to <i>β-actin</i> expression levels. Mean levels (after 8 days) expressed relative to undifferentiated hESCs (shown as one fold) are shown. (E) Model for the miR-181/CARM1/core-pluripotency-factors regulatory loop in the modulation of hESC pluripotency. Pluripotency is maintained in ESCs in part by histone H3 arginine methylation by CARM1 at the <i>Oct4</i>, <i>Nanog</i> and <i>Sox2</i> promoters. The core pluripotency factors also recruit H3K27 methylases to the miR-181c promoter to inhibit its expression. In differentiated hESCs, H3K27 methylation is inhibited due to the reduction of core pluripotency factors, and miR-181 family members are subsequently induced and down-regulate CARM1 activity. H3R17me2 production is eventually stopped, which aggravates the decrease in expression of core pluripotency factors as well as the loss of pluripotency.</p

Supplementary document for Highly sensitive and label-free detection of biotin using liquid crystal-based optofluidic biosensor - 6481542.pdf

Supplementary documen

Neutralizing activity of a third dose of CoronaVac against Omicron subvariants within a 20-month follow-up study

The durability of antibody responses induced by the three-dose of CoronaVac vaccination, especially against SARS-CoV-2 Omicron subvariants, remains unclear. Here in our study, 160 plasma samples from 32 healthy individuals who received three doses of CoronaVac were longitudinally tracked for a period of 20 months. The results showed that a third homologous dose of CoronaVac efficiently increased the SARS-CoV-2 IgG and neutralizing antibody titers and enhanced neutralization activity against Omicron subvariants. The levels of IgG and neutralizing antibody declined from peak levels but remained detectable in most subjects over the course of the next 10–12 months. However, most of the individuals kept neutralizing titers against ancestral Wuhan-Hu-1, while they lost their neutralizing activities against Omicron B.1.1.529, BA.2, BA.4/BA.5, and BA.2.75.2 subvariants at 10–12 months post the third vaccination. Our results suggest that a fourth dose of vaccine may be necessary for uninfected individuals to confer higher neutralization against emerging Omicron subvariants.</p