Fifty whitespotted bamboo sharks (Chiloscyllium playgiosum) of both sexes were used to establish a large capacity variable domain of the new antigen receptor (VNAR) library with a total capacity of over 10⁹ colony-forming units (CFU). It was applied to screen VNARs against human serum albumin (HSA) and human transcription factor EB (TFEB), respectively. Meanwhile, VNAR libraries specific to HSA and TFEB with capacities above 10⁸ CFU were obtained following conventional immunization. These two approaches were systematically studied in terms of VNAR yield and composition. By comparing the VNAR sequences obtained from naïve and antigen-immunized libraries, we found that the complementary-determining region 3 (CDR3) of the former differs in composition from that of the latter. It shares a higher degree of homology with the naïve library. Meanwhile, the binding efficiency assessed by ELISA is also different between the naïve and antigen-immunized libraries. The binding of VNARs from the TFEB-immunized library appeared to surpass that observed with the naïve libraries, whereas the performance of VNARs from the HSA-immunized library indicated that both the immunized and naïve libraries for HSA had positive binding responses in polyclonal and monoclonal ELISA. The results are useful to develop novel diagnostic and therapeutic products based on shark VNARs.

OBJECTIVES: To synthesize a temperature-responsive multimodal motion microrobot (MMMR) using temperature and magnetic field-assisted microfluidic droplet technology to achieve targeted drug delivery and controlled drug release. METHODS: Microfluidic droplet technology was utilized to synthesize the MMMR by mixing gelatin with magnetic microparticles. The microrobot possessed a magnetic anisotropy structure to allow its navigation and targeted drug release by controlling the temperature field and magnetic field. In the experiment, the MMMR was controlled to move in a wide range along a preset path by rotating a uniform magnetic field, and the local circular motion was driven by a planar rotating gradient magnetic field of different frequencies. The MMMR was loaded with simulated drugs, which were released in response to laser heating. RESULTS: Driven by a rotating magnetic field, the MMMR achieved linear motion following a predefined path. The planar gradient rotating magnetic field controlled circular motion of the MMMR with an adjustable radius, utilizing the centrifugal force generated by rotation. The drug-loaded MMMR successfully reached the target location under magnetic guidance, where the gelatin matrix was melted using laser heating for accurate drug release, after which the remaining magnetic particles were removed using magnetic field. CONCLUSIONS The MMMR possesses multimodal motion capabilities to enable precise navigation along a predefined path and dynamic regulation of drug release within the target area, thus having great potential for a wide range of biomedical applications.
Drug Delivery Systems/methods* ; Temperature ; Drug Liberation ; Magnetic Fields ; Robotics ; Gelatin/chemistry* ; Delayed-Action Preparations ; Microfluidics ; Motion

Objective:To study the inhibitory effects and mechanism of Armeniacae Semen Amarum alcohol extract in melanogenesis in zebrafish embryos.Methods:48 HPF healthy zebrafish embryos were randomly divided into blank control group, arbutin group, high-, medium- and low-dosage groups. The blank control group was added with 6 ml embryo culture medium, the arbutin group was added with 8 000 μg/ml arbutin solution 6 ml, and the high-, medium- and low-dosage groups were added with 2 925, 1 950 and 1 300 μg/ml Armeniacae Semen Amarum alcohol extract, respectively. After 24 hours of intervention, the melanin area of zebrafish embryos was observed by stereomicroscope, and the inhibition rate of ethanol extract of bitter almond on tyrosinase (TYR) activity and melanin content in zebrafish embryos were detected by spectrophotometry; the antioxidant activity was evaluated by DPPH radical scavenging capacity, ABTS radical scavenging capacity and CAT activity. RT-qPCR was used to detect the mRNA levels of MITF, Tyr and TYRP2 in zebrafish embryos.Results:Compared with the arbutin group, the low-, medium-, and high-dosage groups showed an increase in melanin area ( P<0.01 or P<0.05), a decrease in TYR activity and melanin content inhibition rate ( P<0.01), and IC50 values for DPPH and ABTS free radical scavenging rates of 23.636 and 3.195 mg/ml, respectively. The CAT activity increased in the medium- and high-dosage groups ( P<0.05 or P<0.01), while the mRNA levels of TYR and MITF decreased in the low-, medium-, and high-dosage groups ( P<0.01). Conclusion:Armeniacae Semen Amarum alcohol extract can effectively inhibit the production of melanin in zebrafish embryos, and its mechanism may be closely related to TYR activity and MITF related signaling pathway.

Objective:To evaluate the implementation of national continuing medical education (CME) base programme about infectious disease control and prevention during 2013-2020, so as to improve the quality management of CME.Methods:According to data from national CME system, Excel and SPSS 27.0 were used to analyze project hosting days, places, teachers, students, project directors and training effect. The counting data were expressed by frequency and percentage [cases (%)], chi-square test was used to make comparison between groups, Mantel-Haenszel chi-square test was used for trend test, and the significance test level of the difference was α = 0.05. Results:A total of 116 projects were conducted from 2013 to 2020, with execution rate of 87.9%(116/132). Most hosting days were 2 to 3 days [57.8% (67/116)]. The majority [65.2% (5 785/8 871)] of trainees had junior and intermediate technical titles. As for trainers, trainers with senior technical titles accounted for 87.6% (758/865), and those with intermediate titles accounted for 12.4% (107/865). Mantel-Haenszel chi-square test showed that there was a linear relationship between the proportion of technical titles and the year ( χ2趋势 = 4.97, P趋势 = 0.026). Project directors almost had senior professional title, and nearly one third of them had the experience of undertaking three or more base projects within 8 years. The top three training modules were parasitic diseases prevention and control, AIDS prevention and control, and viral diseases prevention and control. Trainees were highly satisfied with the training contents. Conclusion:The implementation of the infectious disease prevention and control base programme went well in general from 2013 to 2020. In the future, it’s needed to be demand-oriented, rationally design training programs, enhance the evaluation of training effects, strengthen the construction of public health core capacity, and adopt a strategy of brand development in the process of the infectious disease prevention and control base programme.

The extraordinary advantages associated with mRNA vaccines, including their high efficiency, relatively low severity of side effects, and ease of manufacture, have enabled them to be a promising immunotherapy approach against various infectious diseases and cancers. Nevertheless, most mRNA delivery carriers have many disadvantages, such as high toxicity, poor biocompatibility, and low efficiency in vivo, which have hindered the widespread use of mRNA vaccines. To further characterize and solve these problems and develop a new type of safe and efficient mRNA delivery carrier, a negatively charged SA@DOTAP-mRNA nanovaccine was prepared in this study by coating DOTAP-mRNA with the natural anionic polymer sodium alginate (SA). Intriguingly, the transfection efficiency of SA@DOTAP-mRNA was significantly higher than that of DOTAP-mRNA, which was not due to the increase in cellular uptake but was associated with changes in the endocytosis pathway and the strong lysosome escape ability of SA@DOTAP-mRNA. In addition, we found that SA significantly increased the expression of LUC-mRNA in mice and achieved certain spleen targeting. Finally, we confirmed that SA@DOTAP-mRNA had a stronger antigen-presenting ability in E. G7-OVA tumor-bearing mice, dramatically inducing the proliferation of OVA-specific CLTs and ameliorating the antitumor effect. Therefore, we firmly believe that the coating strategy applied to cationic liposome/mRNA complexes is of potential research value in the field of mRNA delivery and has promising clinical application prospects.

Objective:To provide data support for acquired immunodeficiency syndrome (AIDS) clinical treatment by evaluating the antiviral therapy effect of integrase inhibitors (INIs) and analyzing the drug resistant mutations.Methods:Patients who exposed to INIs more than 1 year were analyzed retrospectively. Virus suppression failure subjects(viral load ≥1 000 copies/ml) were sequenced for drug resistance analysis.Results:Of 408 patients, 12 cases(2.9%) were virologic failure, 8 cases had drug resistance mutations. The proportion of drug resistance was 2.0%(8/408). Among them, only 1 case(0.2%, 1/408) was resistant to INIs with associated mutations: T66I, G118R and E138K; 7 cases(1.7%, 7/408) were resistant to nucleoside reverse transcriptase inhibitors(NRTIs) with major mutations: M184V/I, D67N for NRTIs; 5 cases(1.2%, 5/408) were resistant to non-nucleoside reverse transcriptase inhibitors(NNRTIs) with major mutation: K103N; 1 case(0.2%, 1/408) was resistant to protein inhibitors (PIs), and associated mutations was I50L.1 case was resistant simultaneously to INIs, NRTIs and NNRTIs, Dual-class mutations were observed in 4 cases(3 cases to NRTIs and NNRTIs, 1 case to PIs and NNRTIs).Conclusions:The study result showed that the drug resistance rate of INIs was low in Hunan province, however, the INIs drug resistance surveillance was necessary.

The accurate annotation of transcription start sites(TSSs)and their usage are critical for the mechanistic understanding of gene regulation in different biological contexts.To fulfill this,specific high-throughput experimental technologies have been developed to capture TSSs in a genome-wide manner,and various computational tools have also been developed for in silico pre-diction of TSSs solely based on genomic sequences.Most of these computational tools cast the problem as a binary classification task on a balanced dataset,thus resulting in drastic false positive predictions when applied on the genome scale.Here,we present DeeReCT-TSS,a deep learning-based method that is capable of identifying TSSs across the whole genome based on both DNA sequence and conventional RNA sequencing data.We show that by effectively incorporating these two sources of information,DeeReCT-TSS significantly outperforms other solely sequence-based methods on the precise annotation of TSSs used in different cell types.Furthermore,we develop a meta-learning-based extension for simultaneous TSS annotations on 10 cell types,which enables the identification of cell type-specific TSSs.Finally,we demonstrate the high precision of DeeReCT-TSS on two independent datasets by correlating our predicted TSSs with experimentally defined TSS chromatin states.The source code for DeeReCT-TSS is available at https://github.-com/JoshuaChou2018/DeeReCT-TSS_release and https://ngdc.cncb.ac.cn/biocode/tools/BT007316.

Objective To explore the influence of policy support on attitude of staff toward contracted service in community health service ( CHS) centers in different areas of China. Methods A multi-stage stratified cluster sampling method was used to select participants, and questionnaire survey was conducted among 192 staff (99 from Chengdu and 93 from Xiamen) from 4 community health service centers in Chengdu and Xiamen. In addition, documents on contracted service in CHS were collected and analyzed. EpiData software was used to establish database. Double input and cross-check were implemented. SAS version 9.4 was used for data analyses. Results Differences were found in goals, financial modes and insurances coverage in contracted service in the two areas. Among the 192 participants, 80.7% considered that contracted service was worth to carry out; 90.8% supported the implementation of contracted service in local settings and 78.1% were willing to take more work load on contracted service. In contrary, only 41.6% wished their child work as a family doctor. The main reasons for the negative attitudes were that the performance-based salary system had not been well established or implemented, and there was much more work load that resulted from contracted service. Conclusions The majority of the study subjects held positive attitude toward contracted service, but there were disparities between the four study sites. Relative policy and financial support, proper human resource distribution were critical determinants of contracted service.

Objective To analyze the epidemic status of principal human parasites, so as to provide scientific evidence for making prevention countermeasures for Guangdong Province in the future. Methods In 2015, a survey was performed according to the scheme of "The 3rd National Survey of Principal Human Parasites"as well as the incidence of parasites disease in Guangdong Province,the residents at 48 investiation sites in counties were surveyed. The survey of the soil-transmitted nematodes (Ascariasis, Ancylostoma, Trichuris trichiura and Enterobius vermicularis) and intestinal protozoa were performed based on the ecological regions and stratified by economic and geographic situation.The survey of food borne Clonorchis sinensis was performed along with the soil-transmitted nematodes in rural areas, and it was performed by the sample method of random cluster in cities and towns.The residents in each investigation site as the object, the ovum of the soil-transmitted nematodes, Clonorchis sinensis and other helminths were examed by the modified Kato-Katz method, test tube filter paper was used to identify Hookwormspecies, and the intestinal protozoa was checked by direct smear method. The transparent tape anal swabs method for children aged 3 - 6 years to check Enterobius vermicularis. Results Totally 12 401 residents of 48 survey sites from 22 counties were surveyed, and the total infection rate of intestinal parasites was 8.29%(1 028/12 401). The infection rate of soil-transmitted nematodes was 3.39% (420/12 401),in which the infection rate of Ascariasis, Ancylostoma, Trichuris trichiura and Enterobius vermicularis were 0.52% (64/12 401), 1.89%(234/12 401),0.46%(57/12 401),and 0.52%(65/12 401),respectively.The infection rate of Clonorchis sinensis was 4.90%(608/12 401). Nine hundred and sixty-seven children were tested for eggs of Enterobius vermicularis with the infection rate of 12.41% (120/967). The number of hook larva culture was 153, among them, 140 were hookworm larvae of America and no duodenal hookworm larvae and other nematode species were found.Totally 9 309 residents were tested for intestinal protozoa infection,and the infection rate was 0.31%(29/9 309). Conclusion In Guangdong Province, the infection rate of soil-transmitted nematodes is decreasing while the infection rate of Clonorchis sinensis is still high, and the control work of parasites still should be strengthened especially for food borne parasitic diseases.

Objective:To investigate the potential correlation between miR-223 level in leukocytes and platelet responses to clopidogrel in patients with coronary artery disease.Methods:A cohort of 188 outpatients,who conducted percutaneous coronary intervention (PCI) and received dual antiplatelet therapy,were recruited.The patient's electronic health data were collected,and their blood samples were obtained for measurement of adenosine diphosphate (ADP)-induced whole-blood platelet aggregation.Extreme cases ofplatelet responses to clopidogrel (ultra-vs.non-responder) were measured with miR-223-3p levels in leukocytes.Results:Both groups had similar miR-223-3p levels in leukocytes.There were no significant differences in other demographic and clinical data except for metrics of ADP-induced whole-blood platelet aggregation between the 2 group.Conclusion:MiR-223-3p in peripheral leukocytes is not associated with the altered platelet responses to clopidogrel in PCI outpatients.