Background Vaccination against coronavirus disease 2019 (COVID-19) has become an important public health solution. To date, there has been a lack of data on COVID-19 vaccination willingness, vaccine hesitancy, and vaccination coverage in China since the vaccine has become available. Methods We designed and implemented a cross-sectional, population-based online survey to evaluate the willingness, hesitancy, and coverage of the COVID-19 vaccine among the Chinese population. 8742 valid samples were recruited and classified as the vaccine-priority group (n = 3902; 44.6%) and the non-priority group (n = 4840; 55.4%). Results The proportion of people’s trust in the vaccine, delivery system, and government were 69.0%, 78.0% and 81.3%, respectively. 67.1% of the participants were reportedly willing to accept the COVID-19 vaccination, while 9.0% refused it. 834 (35.5%) reported vaccine hesitancy, including acceptors with doubts (48.8%), refusers (39.4%), and delayers (11.8%). The current coverage was 34.4%, far from reaching the requirements of herd immunity. The predicted rate of COVID-19 vaccination was 64.9%, 68.9% and 81.1% based on the rates of vaccine hesitancy, willingness, and refusal, respectively. Conclusions The COVID-19 vaccine rate is far from reaching the requirements of herd immunity, which will require more flexible and comprehensive efforts to improve the population’s confidence and willingness to vaccinate. It should be highlighted that vaccination alone is insufficient to stop the pandemic; further efforts are needed not only to increase vaccination coverage but also to maintain non-specific prevention strategies.
Cell culture is an essential tool to study cell function. In C. elegans the ability to isolate and culture cells has been limited to embryonically derived cells. However, cells or blastomeres isolated from mixed stage embryos terminally differentiate within 24 hours of culture, thus precluding post-embryonic stage cell culture. We have developed an efficient and technically simple method for large-scale isolation and primary culture of larval-stage cells. We have optimized the treatment to maximize cell number and minimize cell death for each of the four larval stages. We obtained up to 7.8×104 cells per microliter of packed larvae, and up to 97% of adherent cells isolated by this method were viable for at least 16 hours. Cultured larval cells showed stage-specific increases in both cell size and multinuclearity and expressed lineage- and cell type-specific reporters. The majority (81%) of larval cells isolated by our method were muscle cells that exhibited stage-specific phenotypes. L1 muscle cells developed 1 to 2 wide cytoplasmic processes, while L4 muscle cells developed 4 to 14 processes of various thicknesses. L4 muscle cells developed bands of myosin heavy chain A thick filaments at the cell center and spontaneously contracted ex vivo. Neurons constituted less than 10% of the isolated cells and the majority of neurons developed one or more long, microtubule-rich protrusions that terminated in actin-rich growth cones. In addition to cells such as muscle and neuron that are high abundance in vivo, we were also able to isolate M-lineage cells that constitute less than 0.2% of cells in vivo. Our novel method of cell isolation extends C. elegans cell culture to larval developmental stages, and allows use of the wealth of cell culture tools, such as cell sorting, electrophysiology, co-culture, and high-resolution imaging of subcellular dynamics, in investigation of post-embryonic development and physiology.
We aim to evaluate the evolution differences in the incidence and case fatality rate (CFR) of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) Delta and Omicron variants. The average incidence and CFRs were described between different countries. A gamma generalized linear mixed model (GLMM) was used to compare the CFRs of Delta and Omicron variants based on vaccination coverage. Totally, 50 countries were included for analyses. The incidence of coronavirus disease 2019 (COVID‐19) ranged from 0.16/100,000 to 82.95/100,000 during the Delta period and 0.03/100,000 to 440.88/100,000 during the Omicron period. The median CFRs were 8.56 (interquartile range [IQR]: 4.76–18.39) during the Delta period and 3.04 (IQR: 1.87–7.48) during the Omicron period, respectively. A total of 47 out of 50 countries showed decreased CFRs of the Omicron variant with the rate ratio ranging from 0.02 (95% confidence interval [CI]: 0.01–0.03) (in Cambodia) to 0.97 (95% CI: 0.87–1.08) (in Ireland). Gamma GLMM analysis showed that the decreased CFR was largely a result of the decreased pathogenicity of Omicron besides the increased vaccination coverage. The Omicron variant shows a higher incidence but a lower CFR around the world as a whole, which is mainly a result of the decreased pathogenicity by SARS‐CoV‐2's mutation, while the vaccination against SARS‐CoV‐2 still acts as a valuable measure in preventing people from death.
Cell isolation and culture are essential tools for the study of cell function. Isolated cells grown under controlled conditions can be manipulated and imaged at a level of resolution that is not possible in whole animals or even tissue explants. Recent advances have allowed for large-scale isolation and culture of primary C. elegans cells from both embryos and all four larval stages. Isolated cells can be used for single-cell profiling, electrophysiology, and high-resolution microscopy to assay cell autonomous development and behavior. This chapter describes protocols for the isolation and culture of C. elegans embryonic and larval stage cells. Our protocols describe isolation of embryonic and L1 stage cells from nematodes grown on high-density NA22 bacterial plates and isolation of L2 through L4 stage cells from nematodes grown in axenic liquid culture. Both embryonic and larval cells can be isolated from nematode populations within 3 hours and can be cultured for several days. A primer on sterile cell culture techniques is given in the appendices.
The highly expressed Sam68 promotes NF-κB signaling activation, catabolic gene expression and cellular apoptosis in TNF-α-treated chondrocytes, which may provide better insights into the pathophysiology of OA and a potential target for its treatment.
Two strains of Gram-stain-positive, aerobic, non-spore-forming, non-motile, rod-shaped bacteria (designated dk512T and dk508) were isolated from the faeces of Tibetan gazelle (Procapra picticaudata) collected from the Qinghai-Tibet Plateau, PR China. The 16S rRNA gene sequences of the strains showed the highest identity to Microbacterium saccharophilum K-1T (98.0 and 97.9 % similarity, respectively). The phylogenetic analysis based on 16S rRNA gene sequences revealed that dk512T and dk508 were members of the genus Microbacterium , and most closely related to strains Microbacterium mitrae M4-8T and Microbacterium hatanonis FCC-01T. The strains grew optimally on brain-heart infusion (BHI) agar with 5.0 % (v/v) sheep blood at 30 °C, pH 7.0 and with 1.0 % (w/v) NaCl. The genome of type strain dk512T was 3.8 Mb with a G+C content of 70.6 mol%. The average nucleotide identity and digital DNA–DNA hybridization values between strain dk512T and previously characterized Microbacterium species were <95 and <70 %, respectively. In strain dk512T, the detected primary cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0, the main respiratory quinones were MK-9 (37.9 %) and MK-10 (35.7 %), and the polar lipids included diphosphatidylglycerol, phosphatidylglycerol and three unidentified glycolipids. The major cell-wall sugars were rhamnose, ribose and galactose. Alanine, glutamic acid, glycine and ornithine were in the cell-wall peptidoglycan. Based on phenotypic data and phylogenetic inference, these two strains represent a novel species of the genus Microbacterium , named here as Microbacterium wangchenii sp. nov, where dk512T is designated the type strain (=CGMCC 1.16590T=JCM 33494T=KCTC 49313T).
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