In the past few decades, microbubbles were widely used as ultrasound contrast agents in the field of tumor imaging. With the development of research, ultrasound targeted microbubble destruction technology combined with drug-loaded microbubbles can achieve precise drug release and play a therapeutic role. As a micron-scale carrier, microbubbles are difficult to penetrate the endothelial cell space of tumors, and nano-scale drug delivery system—nanobubbles came into being. The structure of the two is similar, but the difference in size highlights the unique advantages of nanobubbles in drug delivery. Based on the classification principle of shell materials, this review summarized micro/nanobubbles used for ultrasound diagnosis or treatment and discussed the possible development directions, providing references for the subsequent development.

Objective To study the acupoint selection and medication rules of acupoint application as an advantageous therapy in treating stroke.Methods The clinical literature from CNKI,Wanfang,CBMdisc,and other databases were searched,and Excel 2013 was used to count the frequency of disease,acupoint selection,and medication,and to analyze the acupoint selection rules using SPSS 25.0 and SPSS Modeler.Results Finally,523 articles were included in the literature,among which,the literature on the treatment of post-stroke constipation with acupoint patch was the most,and the related literature was further screened to analyze the acupoint selection and medication rules,and it was concluded that the most frequently applied acupoints of acupoint application for the treatment of post-stroke constipation were Shenque(RN8),Tianshu(ST25),Zhongwan(RN12),Zusanli(ST36),and the acupoints were mainly taken from conception vessel,stomach meridian and gallbladder meridian,and the core prescriptions were Shenque,Tianshu,Zhongwan,Qihai(RN6),Guanyuan(RN4),Zusanli.For the treatment of post-stroke constipation,acupoint application is often used with Rhei Radix et Rhizoma,Aurantii Fructus Immaturus,Cortex Magnoliae Officinalis,Natrii Sulfas Exsiccatus and Borneolum Syntheticum,among which,warm and cold nature drugs are mainly used,and bitter drugs are most frequently used among five flavors,and most frequently enter to the spleen meridian;and the core prescription is Rhei Radix et Rhizoma,Aurantii Fructus Immaturus,Cortex Magnoliae Officinalis,Natrii Sulfas Exsiccatus and Borneolum Syntheticum.Conclusion Acupoint application is an advantageous treatment for treating post-stroke constipation.The selection of acupoints was based on the conception vessel and stomach meridian,and the medication used were mainly focusing on those with functions of unblocking the bowels and directing qi downward,supplemented by strengthening the spleen and benefiting qi,warming the meridians and nourishing the blood.

Ion concentration polarization (ICP) is an electrical transport phenomenon that occurs at the micro-nano interface under the action of an applied electric field, and the ICP phenomenon can be used to enrich charged particles with high efficiency. The microfluidic chip has the advantages of high precision, high efficiency, easy integration and miniaturization in biochemical analysis, which provides a new solution and technical way for biochemical analysis. In response to the demand for the detection of trace charged target analytes in sample solution, the advantages of high enrichment multiplicity, convenient operation and easy integration of ICP are utilized to provide an effective way for microfluidic biochemical detection. The combination of ICP phenomenon and microfluidic analysis technology has been widely used in the fields of pre-enrichment of charged particles, separation of targets, and detection of target analytes in biochemical analysis. In this paper, the principle of ICP and the microfluidic ICP chip are briefly introduced. Under the action of external electric field, the co-ions pass through the ion-selective nanochannel, the counterions are rejected at the boundary of nanochannel to form a depletion zone, and the charged samples will be enriched at the boundary of the depletion zone. Then the preparation techniques and methods of ICP chips are summarized. Among them, the design of microfluidic channel structure and the preparation and design of nanostructures are emphasized. The basic single-channel structure is analyzed, and the parallel-channel structure as well as the integrated multi-functional microfluidic ICP chip are sorted out and summarized. The preparation methods of nanostructures in ICP chips and their respective advantages and disadvantages are listed, and it is summarized that the current mainstream means are the embedding method and the self-assembly method, and attention is paid to the design of nanostructures preparation methods by both of them. In addition, this paper also discusses how to optimize the enrichment efficiency of ICP chip, through the introduction of multi-field coupling, valve control and other means to achieve the optimization of the enrichment efficiency of target substances. Meanwhile, this paper provides a classified overview of the progress of application of ICP chips in biochemical analysis and detection. ICP chips have been widely used in the research and development of biosensors, which can be used for the enrichment and separation of a variety of analytes including small molecules, nucleic acids, proteins, and cells, etc. By changing the design of microfluidic structures, integrating detection methods and modifying specific antibodies, ICP chips have shown great potential in the fields of rapid enrichment and pre-processing of targets, separation of targets and highly sensitive detection. Finally, it is pointed out that ICP chips are facing challenges in improving enrichment efficiency and selectivity, and solving the problems of fluid control, mixing and transport to match the biological properties of target assay, and that microfluidic ICP chips have been continuously promoting the development of ICP chips through the improvement of materials, chip design and integration of multifunctional units, opening up new possibilities in the field of biochemical analysis methods and applications. It can be seen that microfluidic ICP chips have the advantages of low sample flow rate, good separation and enrichment, high detection efficiency, and easy integration and miniaturization, which have shown good research significance and practical prospects in the field of biochemical detection.

Objective:To construct and purify four respiratory syncytial virus (RSV) PreF proteins through gene sequence design and optimization and evaluate their immunogenicity.Methods:Coronin-1A and T4 trimer protein gene sequences were optimized with Human and CHO codons, and then added to RSV F protein sequence. The above plasmids were transfected into Expi293F cells for protein expression. After purification by nickel column, four trimer proteins were prepared. SDS-PAGE and Western blot were performed for protein identification. BALB/c mice were immunized at week 0 and week 3, and blood samples were collected to measure the activities of binding and neutralizing antibodies in serum.Results:SDS-PAGE and Western blot showed that the four proteins had stable trimer structure. Antigen-antibody affinity test showed that the four trimer proteins had strong affinity with RSV-specific monoclonal antibodies 8897, D25, Motavizumab, AM14 and Palivizumab. The titers of antibodies induced by the two T4 trimers were higher after the initial immunization, while there was a substantial increase in the titers of antibodies induced by Human codon-optimized trimer protein after the second immunization.Conclusions:PreF trimer protein can be prepared by adding any of the two different heterotrimer motifs, and induce effective binding and neutralizing antibodies in mice.

Objective:To compare the immunogenicity of the prefusion (PreF) and postfusion (PostF) conformations of the respiratory syncytial virus (RSV) F protein.Methods:The expression of PreF and PostF recombinant proteins was analyzed by SDS-PAGE and Western blot. The binding affinity between F protein and its specific antibodies was detected by Octet. The binding antibodies and neutralizing antibodies in immune serum were detected after immunizing mice with PreF or PostF recombinant protein.Results:PreF protein was stable in the form of a trimer after modification with higher binding affinity with monoclonal antibodies such as D25, 8897, AM14, Palivizumab and Motavizumab. PostF protein lacked the antigenic site ? and showed a monomer conformation. Besides, it was unable to bind to D25, 8897 and AM14 antibodies. Animal experiments showed that AS01 adjuvant was better than aluminum adjuvant in inducing binding antibodies and neutralizing antibodies against RSV Long strains. The binding antibodies induced by PreF and PostF recombinant proteins had similar binding ability to PreF protein, while the binding antibodies induced by PostF recombinant protein showed stronger binding ability to PostF than to PreF.Conclusions:PreF has more epitopes and the trimer form of PreF recombinant protein after modification is more stable and can induce stronger neutralizing antibodies. Moreover, the immunopotentiating effect of AS01 adjuvant is better than that of aluminum adjuvant. Therefore, stabilization-based trimer structure modification of PreF and the development of adjuvants are crucial for the development of RSV vaccines.

Objective:To explore the cause of inconsistency between the results of trisomy 7 by expanded non-invasive prenatal testing (NIPT-PLUS) and trisomy 18 by prenatal diagnosis.Methods:A pregnant woman who received genetic counseling at Jiaozuo Maternal and Child Health Care Hospital on July 5, 2020 was selected as the study subject. NIPT-PLUS, systematic ultrasound and interventional prenatal testing were carried out. The middle segment and root of umbilical cord, center and edge of the maternal and fatal surface of the placenta were sampled for the validation by copy number variation sequencing (CNV-seq).Results:The result of NIPT-PLUS indicated that the fetus has trisomy 7. Systematic ultrasound has shown multiple malformations including atrioventricular septal defect, horseshoe kidney, and rocker-bottom feet. However, QF-PCR, chromosomal karyotyping analysis, and CNV-seq of amniotic fluid samples all showed that the fetus was trisomy 18. Validation using multiple placental samples confirmed that the middle segment of the umbilical cord contains trisomy 18, the center of the placenta contained trisomy 7, and other placental sites were mosaicism for trisomy 7 and trisomy 18. Notably, the ratio of trisomy 18 became lower further away from the umbilical cord.Conclusion:The false positive results of trisomy 7 and false negative trisomy 18 by NIPT-PLUS was probably due to the existence of placental mosaicism. Strict prenatal diagnosis is required needed aneuploidy is detected by NIPT-PLUS to exclude the influence of placental mosaicisms.

Objective To explore the metastasis rate and related risk factors of T1 stage colonic neuroendocrine tumor(C-NET),and to compare the long-term survival outcomes of patients with non-metastatic(T1N0M0 stage)C-NET after local excision(LE)or radical surgery(RS).Methods Clinical information of 433 patients diagnosed with C-NET in the SEER database from January 1,2004 to December 31,2015 were analyzed.Cox regression was used to analyze the influencing factors of metastasis of C-NET.The patients without metastasis were divided into LE group and RS group,and assigned in a 1∶1 ratio using propensity score matching(PSM)according to gender,age,tumor largest diameter,and infiltration depth,with a caliper value set to 0.02.Kaplan-Meier survival curve was used to analyze 5-year cancer-specific survival(CSS)and overall survival(OS)of patients.Cox regression analysis was used to evaluate the influence of metastasis on survival.Results Among 419 C-NET patients,19(4.52%)had distant metastases.Cox regression analysis showed that 11-20 mm of tumor large diameter(HR=9.264,95%CI 3.322-25.835,P＜0.001),right colon location(HR=0.116,95%CI 0.042-0.321,P＜0.001),and submucosal invasion(HR=5.842,95%CI 1.858-18.371,P=0.003)were independent risk factors for distant metastasis of T1 stage C-NET.The 5-year OS rates of non-metastatic and metastatic patients were 94.5%and 47.4%,respectively(χ2=79.762,P＜0.001),and their 5-year CSS rates were 99.5%and 55.7%,respectively(χ2=164.604,P＜0.001).Before PSM,the 5-year OS rates of non-metastatic C-NET patients after LE and RS were 95.8%and 90.1%(χ2=2.679,P=0.063),and the 5-year CSS rates were 100.0%and 97.2%(χ2=0.579,P=0.038);after PSM,the 5-year OS rates of non-metastatic patients after LE and RS were 96.8%and 92.1%(χ2=3.606,P=0.058),and the 5-year CSS rates were 100.0%and 98.5%(χ2=1.015,P=0.314).After PSM,there was no significant difference in the 5-year OS and CSS of patients with defferent tumor location,tumor large diameter,or submucosal invasion between the LE and RS groups.Conclusions 11-20 mm of tumor diameter,right colon location,and submucosal invasion might be independent risk factors for distant metastasis of T1 stage C-NET,and LE could be an appropriate treatment option for non-metastatic C-NET.

Acute liver injury (ALI) is one of the common severe diseases in clinic, which is characterized by redox imbalance and inflammatory storm. Untimely treatment can easily lead to liver failure and even death. Rosmarinic acid (RA) has been proved to have anti-inflammatory and antioxidant activity, but it is not clear how to protect ALI through antioxidation and inhibition of inflammation. Therefore, this study explored the therapeutic effect and molecular mechanism of RA on ALI through in vitro and in vivo experiments. In the mouse ALI model, the effects of RA on liver function and inflammatory indexes were studied, the pathological changes of liver were observed by HE, the effect of RA on reactive oxygen species in liver was detected by fluorescence method, and the level of F4/80 in liver tissue was detected by immunohistochemical method. The levels of thioredoxin interacting protein (TXNIP), NOD-like receptor protein 3 (NLRP3) and cysteinyl aspartate specific proteinase-1 (CASPASE-1) in liver tissue were measured by Western blot. All animal welfare and experimental procedures follow the rules of the Animal Ethics Committee of Southwest Minzu University. In vitro, human hepatoma cell line HepG2 was used to establish the model of oxidative damage induced by H₂O₂. The cell viability was detected by CCK-8 method. The level of interleukin-1β (IL-1β) in the supernatant was detected by enzyme linked immunosorbent assay (ELISA), the activity of lactatede hydrogenase (LDH) was detected by LDH kit, and the level of ROS was detected by fluorescence probe DCFH-DA labeling. The mRNA expression of Txnip, Nlrp3, Caspase 1, Il1β was detected by real-time fluorescence quantitative PCR (qPCR), and the protein levels of nuclear factor erythroid-2 related factor 2 (NRF2), TXNIP, NLRP3 and CASPASE-1 were measured by Western blot. The results showed that compared with the model group, the degree of liver swelling, tissue injury, liver function index in RA group were significantly lower than those in model group. And RA significantly attenuated the increases of ROS in liver tissue. The expression levels of TXNIP, NLRP3 and CASPASE-1 in liver tissue were significantly lower than those in model group. Additionally, RA inhibited the expressions of F4/80 and IL-1β. In vitro experiment, compared with model group, RA effectively inhibited the secretion of IL-1β and LDH. The level of ROS also decreased significantly. RA inhibited the mRNA expressions of Txnip, Caspase 1, Il1β. Furthermore, RA significantly increased the expression level of NRF2 protein in nucleus, and decreased the expression level of TXNIP and NLRP3 protein. Specifically, with the addition of ML385, the effect of RA on NRF2, TXNIP, NLRP3, CASPASE-1 protein expression was reversed. Collectively, these findings suggested that RA may inhibit the production of ROS by promoting NRF2 nuclear transfer, and then reduce the activation of NLRP3 inflammatory bodies by TXNIP, reduce cell death and inflammatory response to prevent the liver injury.

Cervical spondylotic radiculopathy(CSR)is a condition caused by the degeneration of cervical intervertebral discs and facet joints,primarily manifesting as the pain,sensory abnormalities,and motor dysfunction in the cervical nerve innervation area of neck,shoulder,and upper limb.For the treatment of CSR,tendon-bone syndrome differentiation in traditional Chinese medicine often faces the issues of conceptual confusion and non-standard syndrome differentiation.Based on the traditional tendon-bone syndrome differentiation and by integrating modern anatomical insights,Professor LIN Ding-Kun,an esteemed scholar of Traditional Chinese Medicine,proposed a classification system for the cervical spine that includes the categories of tendons,joints,bones and marrow.This paper explored the thoughts of Professor LIN for the tendon-bone syndrome differentiation of CSR,summarized the targets of manual therapy,and proposed the four kinds of pathological changes such as tendon overstrain,joint dislocation,bone lesion,and marrow injury,as well as the four techniques of traditional Chinese medicine manipulations,i.e.relaxation of tendons,reduction of joints,protection of marrow,and treatment of bones.The aim is to improve the syndrome-differentiation and treatment for CSR with orthopedic and traumatologic manipulations,and to provide reference for clinical practice.

Acute myocardial infraction(AMI)has now become a threat to human health worldwide.Therefore,it is urgent to construct the standardized platform of integrated traditional Chinese and western medicine diagnosis and treatment for AMI.The Intensive Care Unit(ICU)team of the Second Clinical Medical School of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Chinese Medicine)has taken up the mission of exploring the prevention and treatment of AMI with traditional Chinese medicine and promoting and the construction of the diagnosis and treatment system of integrated traditional Chinese and western medicine for AMI.The team pioneered the trinity mode of'saving heart,treating heart and nourishing heart'for AMI,focusing on the construction of standardized platform of integrated traditional Chinese and western medicine diagnosis and treatment for AMI,creating the key technology system of Yiqi Huoxue Huatan method(the therapy mainly for replenishing qi,activating blood and dissolving phlegm)for AMI and putting it into the practice,which highlighted the achievements in the construction of the standardized platform.