Understanding Chromium Slag Recycling with Sintering-Ironmaking Processes: Influence of Cr2O3 on the Sinter Microstructure and Mechanical Properties of the Silico-Ferrite of Calcium and Aluminum (SFCA).

Chromium slag is a solid waste of chromium salt production, which contains highly toxic Cr(VI) and significant amounts of valuable metals, such as Fe and Cr. Recycling chromium slag as a raw sintering material in sintering-ironmaking processes can simultaneously reduce toxic Cr(VI) and recover valuable metals. A micro-sintering experiment, compressive strength test, microhardness test, and first-principles calculation are performed to investigate the influence of Cr2O3 on the sintering microstructure and mechanical properties of the silico-ferrite of calcium and aluminum (SFCA) in order to understand the basis of the sintering process with chromium slag addition. The results show that the microstructure of SFCA changes from blocky to interwoven, with further increasing Cr2O3 content from 0 wt% to 3 wt%, and transforms to blocky with Cr2O3 content increasing to 5 wt%. Cr2O3 reacts with Fe2O3 to form (Fe1-xCrx)2O3 (0 ≤ x ≤ 1), which participates in forming SFCA. With the increase in Cr doping concentrations, the hardness of SFCA first decreases and then increases, and the toughness increases. When Cr2O3 content increases from 0 wt% to 3 wt%, the SFCA microhardness decreases and the compressive strength of the sintered sample increases. Further increasing Cr2O3 contents to 5 wt%, the SFCA microhardness increases, and the compressive strength of sintered sample decreases.

A new illudane sesquiterpene from the edible fungus Pholiota nameko.

Fungi have different genetic expression abilities and biosynthetic pathways under different cultivation conditions, which can produce various secondary metabolites. The "one strain many compounds" strategy is used to activate silent biosynthetic genes of fungi to produce various compounds, which is an effective method. In order to discover various new compounds in the edible fungus Pholiota nameko, a fermentation strategy involving precursor feeding and enzyme inhibitor addition has been employed. A new illudane sesquiterpene (1), along with one known indole diterpenoid alkaloid, cladosporine A (2) were isolated from the extracts of liquid culture of P. nameko. The new compound was identified by combination of 1D and 2D NMR, MS, optical rotation, and ECD calculations. We conducted experiments on the cytotoxicity of all isolated compounds on three cancer cell lines, but we did not observe any significant cytotoxicity (IC50 > 40 µM).

Does entrepreneurship education in China develop entrepreneurial intention? the role of self-efficacy and experience.

Entrepreneurship education has attracted much attention in recent years. However, the relationship between entrepreneurship education and entrepreneurial intention has not achieved an agreement yet. To reconcile these conflicting conclusions, we explore the effect of entrepreneurship education on entrepreneurial intention from the content of the entrepreneurship education programs and different types of individuals who have participated in the program. Leveraging the self-efficacy theory and event system theory, we examine the mediation of entrepreneurial self-efficacy from five dimensions and the moderation of entrepreneurial experience. The sample of this study comprised 243 individuals who participated in entrepreneurship education in China (female = 40.3%, The majority of responders with an age range from 21 to 30 years). The results reveal that entrepreneurship education has a significantly positive influence on entrepreneurial intention (ß = 0.331, p < 0.001). Entrepreneurial self-efficacies in searching (ß = 0.382, p<0.001), planning (ß = 0.249, p<0.001), and marshaling (ß = 0.134, p<0.05) play mediating roles in the relationship between entrepreneurship education and entrepreneurial intention. We also find that entrepreneurial experience negatively moderates the relationship between entrepreneurship education and entrepreneurial intention (ß = -0.212, p<0.05). The results have implications for entrepreneurship education scholars and policymakers in China.

Molecular Characterization of Two Wamide Neuropeptide Signaling Systems in Mollusk Aplysia.

Neuropeptides with the C-terminal Wamide (Trp-NH2) are one of the last common ancestors of peptide families of eumetazoans and play various physiological roles. In this study, we sought to characterize the ancient Wamide peptides signaling systems in the marine mollusk Aplysia californica, i.e., APGWamide (APGWa) and myoinhibitory peptide (MIP)/Allatostatin B (AST-B) signaling systems. A common feature of protostome APGWa and MIP/AST-B peptides is the presence of a conserved Wamide motif in the C-terminus. Although orthologs of the APGWa and MIP signaling systems have been studied to various extents in annelids or other protostomes, no complete signaling systems have yet been characterized in mollusks. Here, through bioinformatics, molecular and cellular biology, we identified three receptors for APGWa, namely, APGWa-R1, APGWa-R2, and APGWa-R3. The EC50 values for APGWa-R1, APGWa-R2, and APGWa-R3 are 45, 2100, and 2600 nM, respectively. For the MIP signaling system, we predicted 13 forms of peptides, i.e., MIP1-13 that could be generated from the precursor identified in our study, with MIP5 (WKQMAVWa) having the largest number of copies (4 copies). Then, a complete MIP receptor (MIPR) was identified and the MIP1-13 peptides activated the MIPR in a dose-dependent manner, with EC50 values ranging from 40 to 3000 nM. Peptide analogs with alanine substitution experiments demonstrated that the Wamide motif at the C-terminus is necessary for receptor activity in both the APGWa and MIP systems. Moreover, cross-activity between the two signaling systems showed that MIP1, 4, 7, and 8 ligands could activate APGWa-R1 with a low potency (EC50 values: 2800-22,000 nM), which further supported that the APGWa and MIP signaling systems are somewhat related. In summary, our successful characterization of Aplysia APGWa and MIP signaling systems represents the first example in mollusks and provides an important basis for further functional studies in this and other protostome species. Moreover, this study may be useful for elucidating and clarifying the evolutionary relationship between the two Wamide signaling systems (i.e., APGWa and MIP systems) and their other extended neuropeptide signaling systems.

Identification of three elevenin receptors and roles of elevenin disulfide bond and residues in receptor activation in Aplysia californica.

Neuropeptides are ubiquitous intercellular signaling molecules in the CNS and play diverse roles in modulating physiological functions by acting on specific G-protein coupled receptors (GPCRs). Among them, the elevenin signaling system is now believed to be present primarily in protostomes. Although elevenin was first identified from the L11 neuron of the abdominal ganglion in mollusc Aplysia californica, no receptors have been described in Aplysia, nor in any other molluscs. Here, using two elevenin receptors in annelid Platynereis dumerilii, we found three putative elevenin GPCRs in Aplysia. We cloned the three receptors and tentatively named them apElevR1, apElevR2, and apElevR3. Using an inositol monophosphate (IP1) accumulation assay, we demonstrated that Aplysia elevenin with the disulfide bond activated the three putative receptors with low EC50 values (ranging from 1.2 to 25 nM), supporting that they are true receptors for elevenin. In contrast, elevenin without the disulfide bond could not activate the receptors, indicating that the disulfide bond is required for receptor activity. Using alanine substitution of individual conserved residues other than the two cysteines, we showed that these residues appear to be critical to receptor activity, and the three different receptors had different sensitivities to the single residue substitution. Finally, we examined the roles of those residues outside the disulfide bond ring by removing these residues and found that they also appeared to be important to receptor activity. Thus, our study provides an important basis for further study of the functions of elevenin and its receptors in Aplysia and other molluscs.

Characterization of an Aplysia vasotocin signaling system and actions of posttranslational modifications and individual residues of the ligand on receptor activity.

The vasopressin/oxytocin signaling system is present in both protostomes and deuterostomes and plays various physiological roles. Although there were reports for both vasopressin-like peptides and receptors in mollusc Lymnaea and Octopus, no precursor or receptors have been described in mollusc Aplysia. Here, through bioinformatics, molecular and cellular biology, we identified both the precursor and two receptors for Aplysia vasopressin-like peptide, which we named Aplysia vasotocin (apVT). The precursor provides evidence for the exact sequence of apVT, which is identical to conopressin G from cone snail venom, and contains 9 amino acids, with two cysteines at position 1 and 6, similar to nearly all vasopressin-like peptides. Through inositol monophosphate (IP1) accumulation assay, we demonstrated that two of the three putative receptors we cloned from Aplysia cDNA are true receptors for apVT. We named the two receptors as apVTR1 and apVTR2. We then determined the roles of post-translational modifications (PTMs) of apVT, i.e., the disulfide bond between two cysteines and the C-terminal amidation on receptor activity. Both the disulfide bond and amidation were critical for the activation of the two receptors. Cross-activity with conopressin S, annetocin from an annelid, and vertebrate oxytocin showed that although all three ligands can activate both receptors, the potency of these peptides differed depending on their residue variations from apVT. We, therefore, tested the roles of each residue through alanine substitution and found that each substitution could reduce the potency of the peptide analog, and substitution of the residues within the disulfide bond tended to have a larger impact on receptor activity than the substitution of those outside the bond. Moreover, the two receptors had different sensitivities to the PTMs and single residue substitutions. Thus, we have characterized the Aplysia vasotocin signaling system and showed how the PTMs and individual residues in the ligand contributed to receptor activity.

Efficient extraction, excellent activity, and microencapsulation of flavonoids from Moringa oleifera leaves extracted by deep eutectic solvent.

A deep eutectic solvent (choline chloride (ChCl)-urea) was chosen to extract flavonoids from Moringa oleifera leaves (FMOL), the condition of extraction was tailor-made, under the optimal extraction conditions (material-to-liquid ratio of 1:60 g/mL, extraction time of 80 min, extraction temperature of 80 °C), the highest extraction efficiency reached 63.2 ± 0.3 mg R/g DW, and nine flavonoids were identified. Then, the biological activities including antioxidant activities, antibacterial activities, and anti-tumor activities were systematically studied. FMOL was superior to positive drugs in terms of antioxidant activity. As to DPPH investigation, the IC50 of FMOL and Vc were 64.1 ± 0.7 and 176.1 ± 2.0 µg/mL; for the ABTS, the IC50 of FMOL and Vc were 9.5 ± 0.3 and 38.2 ± 1.2 µg/mL, the FRAP value of FMOL and Vc were 15.5 ± 0.6 and 10.2 ± 0.4 mg TE/g, and ORAC value of FMOL and Vc were 4687.2 ± 102.8 and 3881.6 ± 98.6 µmol TE/g. The bacteriostatic (MICs were ≤ 1.25 mg/mL) activities of FMOL were much better than propyl p-hydroxybenzoate. Meanwhile, FMOL had comparable inhibitory activity with genistein on tumor cells, IC50 was 307.8 µg/mL, and could effectively induce apoptosis in HCT116. Microcapsules were prepared with xylose-modified soybean protein isolate and gelatin as wall materials; after that, the intestinal release of modified FMOL microcapsules was 86 times of free FMOL. Therefore, this study confirmed that FMOL extracted with ChCl/urea has rich bioactive components, and microencapsulated FMOL has potential application in food industry. Supplementary Information: The online version contains supplementary material available at 10.1007/s13399-023-03877-8.

Interface modified BTO@PS-co-mah/PS composite dielectrics with enhanced breakdown strength and ultralow dielectric loss.

Dielectrics of the polymer-matrix composite are considered to present combined advantages from both the polymer matrix and inorganic fillers. However, the breakdown strength, as well as energy density, is not effectively enhanced due to the poor compatibility between the organic and inorganic components. Herein, polymer composites derived from polystyrene (PS) and barium titanate (BTO) are proposed and beneficial interface modification by poly(styrene-co-maleic anhydride) (PS-co-mah) is conducted to improve compatibility between the inorganic filler and polymer matrix. The results show that the BTO@PS-co-mah/PS composites, in which the interfacial layer of PS-co-mah would undergo chemical reactions with the aminated BTO and blend PS matrix with excellent physical compatibility, exhibit enhanced breakdown strength and declined dielectric loss compared with both pure PS and BTO/PS without interfacial modulation. Particularly, the BTO@PS-co-mah/PS composite with 5 wt% filler content indicates optimized performance with an E b of 507 MV m-1 and tan δ of 0.085%. It is deduced that the deep energy traps introduced by the PS-co-mah layer would weaken the local electric field and suppress the space charge transporting so as to optimize the performance of composites. Consequently, the interfacial-modified BTO@PS-co-mah/PS would present great potential for applications, such as film capacitors.

Integrated exploration of experimentation and molecular simulation in ester-containing polyimide dielectrics.

With the growing development of film capacitors in various applications, the requirements for polymer dielectrics have increased accordingly. In this work, a series of ester-cotaining polyimide (EPI) dielectrics were designed and fabricated. Futhermore, integrated exploration of experimentation and molecular simulation is proposed to achieve polymer dielectrics with advanced comprehensive performance, as well as to analyze the dielectric mechanism in-depth. The EPIs show superior thermal resistance and dielectric properties. A Weibull breakdown strength of 440-540 MV m-1, permittivity of 3.52-3.85, dissipation factor of 0.627-0.880% and theoretical energy density of 3.13-4.90 J cm-3 were obtained for the EPIs. The relationship between microscopic parameters and dielectric behavior was investigated in detail. According to the experimental and calculated results, there is close correlation between dipolar moment density (µ/V vdw) and dielectric permittivity (ε r). It is deduced that the integrated research of experiments and molecular simulation would be an effective strategy to reveal the dielectric mechanism as well as assist in the molecular design of polymer dielectrics.

Significantly Improved Dielectric Performance of All-Organic Parylene/Polyimide/Parylene Composite Films with Sandwich Structure.

The development of novel polymer dielectrics with enhanced dielectric performance is a great challenge for application of film capacitors in modern electronics and electrical systems. Herein, an innovative approach of chemical vapor deposition polymerization technology is proposed to prepare the all-organic sandwich structured parylene/polyimide/parylene (Py/PI/Py) composite films by employing poly(chloro-para-xylylene) (parylene C) as the outer layers and polyimide (PI) as the inner layer. The Py/PI/Py composites exhibit superior thermal resistance and outstanding mechanical properties. Moreover, thanks to the interfacial effect which contributes to reinforcing the dielectric response and the thickness effect which facilitates improving the breakdown strength, the dielectric performance of Py/PI/Py composites has been enhanced significantly. Accordingly, dielectric constant of 4.52-5.09, dissipation factor of 0.21-1.01%, and breakdown strength of 307-460 MV m-1 are achieved. Besides, notable energy storage performance is also obtained in Py/PI/Py composite dielectrics. Consequently, this novel application of chemical vapor deposition polymerization method in preparing all-organic multilayered polymer composite films with sandwich structure shows promising potential in film capacitor applications in harsh conditions.

Hepatic vagotomy blunts liver regeneration after hepatectomy by downregulating the expression of interleukin-22.

BACKGROUND: Rapid regeneration of the residual liver is one of the key determinants of successful partial hepatectomy (PHx). At present, there is a lack of recognized safe, effective, and stable drugs to promote liver regeneration. It has been reported that vagus nerve signaling is beneficial to liver regeneration, but the potential mechanism at play here is not fully understood. AIM: To explore the effect and mechanism of hepatic vagus nerve in liver regeneration after PHx. METHODS: A PHx plus hepatic vagotomy (Hv) mouse model was established. The effect of Hv on liver regeneration after PHx was determined by comparing the liver regeneration levels of the PHx-Hv group and the PHx-sham group mice. In order to further investigate the role of interleukin (IL)-22 in liver regeneration inhibition mediated by Hv, the levels of IL-22 in the PHx-Hv group and the PHx-sham group was measured. The degree of liver injury in the PHx-Hv group and the PHx-sham group mice was detected to determine the role of the hepatic vagus nerve in liver injury after PHx. RESULTS: Compared to control-group mice, Hv mice showed severe liver injury and weakened liver regeneration after PHx. Further research found that Hv downregulates the production of IL-22 induced by PHx and blocks activation of the signal transducer and activator of transcription 3 (STAT3) pathway then reduces the expression of various mitogenic and anti-apoptotic proteins after PHx. Exogenous IL-22 reverses the inhibition of liver regeneration induced by Hv and alleviates liver injury, while treatment with IL-22 binding protein (an inhibitor of IL-22 signaling) reduce the concentration of IL-22 induced by PHx, inhibits the activation of the STAT3 signaling pathway in the liver after PHx, thereby hindering liver regeneration and aggravating liver injury in PHx-sham mice. CONCLUSION: Hv attenuates liver regeneration after hepatectomy, and the mechanism may be related to the fact that Hv downregulates the production of IL-22, then blocks activation of the STAT3 pathway.

Fabrication of Cu@Sn TLPS joint for high temperature power electronics application.

In this study, a novel Cu@Sn TLPS joint was fabricated for high-temperature power electronics application. Cu@Sn core-shell composite powder was firstly prepared by a methylate electroplating method, and then pressed into a preformed sheet. The Cu@Sn preform was reflowed at 250 ∼ 280 °C for 40 min under the pressure of 0.1 × 10-3 MPa, and the resulting bondline can withstand high temperatures up to 600 °C. During the process, the Sn layer was transformed to Cu3Sn, and the Cu3Sn surrounded the outside of the residual Cu particles. The joint characteristics were controlled by size gradation of Cu particles, the ratio of Cu/Sn, preform forming pressure and TLPS process. The joint shear strength was no less than 48 MPa after aging at 400 °C for 1000 h. Young's modulus and hardness were 98.35 GPa and 2.62 GPa, respectively, which are much lower than the pure Cu3Sn joint. The electrical resistivity and thermal conductivity of the joint were 5.1 µΩ cm and 148 W m-1 K-1, respectively. It is superior to pure Cu3Sn joints and the other Cu/Sn system TLPS joints. The high shear strength, high thermal conductivity and high melting temperature demonstrate that Cu@Sn TLPS joint is a promising interconnect technology for high power density modules.

AI protein structure prediction-based modeling and mutagenesis of a protostome receptor and peptide ligands reveal key residues for their interaction.

The protostome leucokinin (LK) signaling system, including LK peptides and their G protein-coupled receptors, has been characterized in several species. Despite the progress, molecular mechanisms governing LK peptide-receptor interactions remain to be elucidated. Previously, we identified a precursor protein for Aplysia leucokinin-like peptides (ALKs) that contains the greatest number of amidated peptides among LK precursors in all species identified so far. Here, we identified the first ALK receptor from Aplysia, ALKR. We used cell-based IP1 activation assays to demonstrate that two ALK peptides with the most copies, ALK1 and ALK2, activated ALKR with high potencies. Other endogenous ALK-derived peptides bearing the FXXWX-amide motif also activated ALKR to various degrees. Our examination of cross-species activity of ALKs with the Anopheles LK receptor was consistent with a critical role for the FXXWX-amide motif in receptor activity. Furthermore, we showed, through alanine substitution of ALK1, the highly conserved phenylalanine (F), tryptophan (W), and C-terminal amidation were each essential for receptor activation. Finally, we used an artificial intelligence-based protein structure prediction server (Robetta) and Autodock Vina to predict the ligand-bound conformation of ALKR. Our model predicted several interactions (i.e., hydrophobic interactions, hydrogen bonds, and amide-pi stacking) between ALK peptides and ALKR, and several of our substitution and mutagenesis experiments were consistent with the predicted model. In conclusion, our results provide important information defining possible interactions between ALK peptides and their receptors. The workflow utilized here may be useful for studying other ligand-receptor interactions for a neuropeptide signaling system, particularly in protostomes.

Comparison of mitochondrial genetic variation of Taenia hydatigena cysticerci from China and Mongolia.

Parasitic infection is one of the many challenges facing livestock production globally. Cysticercosis tenuicollis is a common parasitic disease in domestic and wild ruminants (intermediate host) caused by the larval stage of Taenia hydatigena that primarily infects dogs (definitive host). Although genetic studies on this parasite exist, only a few describe the genetic variation of this parasite in Mongolia. Our aim was thus, to identify the mitochondrial differences in ovine isolates of Cysticercus tenuicollis entering China from Mongolia and comparison with existing Chinese isolates from sheep and goats based on the recently described PCR-RFLP method and mitochondrial genes of NADH dehydrogenase subunit 4 (nad4) and the NADH dehydrogenase subunit 5 (nad5). Sixty-nine isolates were collected during routine veterinary meat inspections from sheep that originated from Mongolia, at the modern slaughterhouses in Erenhot City, Inner Mongolia. Additional 114 cysticerci were also retrieved from sheep and goats from northern (Inner Mongolia Autonomous Region, Ningxia Hui Autonomous Region, and Gansu Province), western (Tibet Autonomous Region), and southern (Jiangxi Province and Guangxi Province) China. The PCR-RFLP approach of the nad5 showed nine mitochondrial subclusters A1, A2, A3, A5, A8, A9, A10, A11, and B of T. hydatigena isolates from sheep and goats from Mongolia and China. Meanwhile, haplogroup A1 RFLP profile was more widespread than other variants. These data supplements existing information on the molecular epidemiology of T. hydatigena in China and Mongolia and demonstrate the occurrence of similar genetic population structures in both countries.

Toward high-power-density laser-driven lighting: enhancing heat dissipation in phosphor-in-glass film by introducing h-BN.

In this work,  hexagonal boron nitride (h-BN) nanocrystals as functional additives in a phosphor-in-glass film are shown to substantially increase the luminous performance driven by a blue laser. Microstructural and spectroscopic studies reveal that h-BN particles distributed over the whole glass matrix build in situ a local heat conductive path which effectively accelerates heat dissipation and so greatly relieves the "thermal run-away effect". The developed composite material with fine thermal manipulation may be a promising phosphor color converter for high-power-density laser-driven lighting.

High-affinity anti-Arc nanobodies provide tools for structural and functional studies.

Activity-regulated cytoskeleton-associated protein (Arc) is a multidomain protein of retroviral origin with a vital role in the regulation of synaptic plasticity and memory formation in mammals. However, the mechanistic and structural basis of Arc function is poorly understood. Arc has an N-terminal domain (NTD) involved in membrane binding and a C-terminal domain (CTD) that binds postsynaptic protein ligands. In addition, the NTD and CTD both function in Arc oligomerisation, including assembly of retrovirus-like capsids involved in intercellular signalling. To obtain new tools for studies on Arc structure and function, we produced and characterised six high-affinity anti-Arc nanobodies (Nb). The CTD of rat and human Arc were both crystallised in ternary complexes with two Nbs. One Nb bound deep into the stargazin-binding pocket of Arc CTD and suggested competitive binding with Arc ligand peptides. The crystallisation of the human Arc CTD in two different conformations, accompanied by SAXS data and molecular dynamics simulations, paints a dynamic picture of the mammalian Arc CTD. The collapsed conformation closely resembles Drosophila Arc in capsids, suggesting that we have trapped a capsid-like conformation of the human Arc CTD. Our data obtained with the help of anti-Arc Nbs suggest that structural dynamics of the CTD and dimerisation of the NTD may promote the formation of capsids. Taken together, the recombinant high-affinity anti-Arc Nbs are versatile tools that can be further developed for studying mammalian Arc structure and function, as well as mechanisms of Arc capsid formation, both in vitro and in vivo. For example, the Nbs could serve as a genetically encoded tools for inhibition of endogenous Arc interactions in the study of neuronal function and plasticity.

Case Report: Chimeric Antigen Receptor T Cells Induced Late Severe Cytokine Release Syndrome.

Background: Severe cytokine release syndrome (sCRS) has emerged as an adverse complication in the early period of chimeric antigen receptor T cell (CART) therapy, while whether sCRS occurs in the late period remains unknown. Here, we reported two patients with late sCRS. Case Presentation: Case 1 was a 34-year-old female with refractory Philadelphia chromosome-positive B cell acute lymphoblastic leukemia. She achieved complete remission (CR) but experienced grade III CRS and hemophagocytic lymphohistiocytosis (HLH) 41 days after CD19-targeted CART (CART19) cells and CD22-targeted CART (CART22) cells infusion. Ineffective to tocilizumab and HLH-94 protocol (dexamethasone and etoposide), she died of a cerebral hemorrhage on day 55 after CART therapy. Case 2 was a 38-year-old male with IgG kappa multiple myeloma. He received autologous BCMA-targeted CART (BCMA-CART) therapy 4 months after HLA-matched sibling (sister) donor transplantation and developed grade III CRS 163 days after CART administration, characterized by fever, hypotension, and skin lesions. Effective to methylprednisolone and tocilizumab, his clinical response persisted for over 6.0 months. Conclusion: Severe CRS could occur in the late period after CART therapy as re-expansion of CART cells possessed the potential risk for late sCRS.

The Persistence and Change in China's Healthcare Insurance Reform: Clues from Fiscal Subsidy Policies Made for Settling COVID-19 Patients' Medical Costs.

Background: The system studied in this article is set in the development and reform of China's healthcare insurance, the time mentioned in this article is during the COVID-19 period and the issues are about healthcare insurance fiscal subsidy policies for COVID-19 patients' medical costs. Methods: Method of comparison and method of case study are used. This article compares the healthcare insurance systems, fiscal subsidy policies during the COVID-19 period and takes Yulin's healthcare insurance reform and Nanjing healthcare insurance administration's measures to fight against COVID-19 pandemic as specific cases to analyze. Aim: This article presents the advantages of the coverage of China's healthcare insurance as well as the insufficiency of the commercial healthcare insurance and market-oriented medical institutions to show the persistence and change in China's healthcare insurance reform. Conclusion: Soon after the outbreak of COVID-19 pandemic, the National Healthcare Security Administration and the Ministry of Finance of PRC jointly issued an announcement on how to guarantee the medical security of the novel coronavirus pneumonia pandemic. More specific policies were made by each province to secure that the total medical costs of the COVID-19 confirmed and suspected patients were almost borne by healthcare insurance and government finance subsidy. These policies reveal two basic points ahead China's healthcare reform: one is that China shall persist in positively promoting the universal healthcare insurance coverage program, which will break down barriers between urban and rural areas; the other is that the commercial medical insurance shall be a supplementary to social healthcare insurance and the market-oriented medical institutions shall be encouraged. The two basic points stand for the persistence and change in China's healthcare reform. China's healthcare insurance reform should accord with the requirement in specific age and stage.

Development and Validation of Arc Nanobodies: New Tools for Probing Arc Dynamics and Function.

Activity-regulated cytoskeleton-associated (Arc) protein plays key roles in long-term synaptic plasticity, memory, and cognitive flexibility. However, an integral understanding of Arc mechanisms is lacking. Arc is proposed to function as an interaction hub in neuronal dendrites and the nucleus, yet Arc can also form retrovirus-like capsids with proposed roles in intercellular communication. Here, we sought to develop anti-Arc nanobodies (ArcNbs) as new tools for probing Arc dynamics and function. Six ArcNbs representing different clonal lines were selected from immunized alpaca. Immunoblotting with recombinant ArcNbs fused to a small ALFA-epitope tag demonstrated binding to recombinant Arc as well as endogenous Arc from rat cortical tissue. ALFA-tagged ArcNb also provided efficient immunoprecipitation of stimulus-induced Arc after carbachol-treatment of SH-SY5Y neuroblastoma cells and induction of long-term potentiation in the rat dentate gyrus in vivo. Epitope mapping showed that all Nbs recognize the Arc C-terminal region containing the retroviral Gag capsid homology domain, comprised of tandem N- and C-lobes. ArcNbs E5 and H11 selectively bound the N-lobe, which harbors a peptide ligand binding pocket specific to mammals. Four additional ArcNbs bound the region containing the C-lobe and C-terminal tail. For use as genetically encoded fluorescent intrabodies, we show that ArcNbs fused to mScarlet-I are uniformly expressed, without aggregation, in the cytoplasm and nucleus of HEK293FT cells. Finally, mScarlet-I-ArcNb H11 expressed as intrabody selectively bound the N-lobe and enabled co-immunoprecipitation of full-length intracellular Arc. ArcNbs are versatile tools for live-cell labeling and purification of Arc, and interrogation of Arc capsid domain specific functions.