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    Development and application of intestine-on-a-chip technology
    HU Liang, PEI Xuetao, LI Yanhua
    2023, 47(8): 561-565. https://doi.org/10.7644/j.issn.1674-9960.2023.08.001
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    The intestine-on-a-chip is a cutting-edge and in vitro research model of human intestines that has gained much attention in recent years. In comparison to traditional static culture models of intestinal epithelial cells, the intestine-on-a-chip offers numerous advantages. It accurately mimics the morphological structure of the intestine in vivo, allowing for the simulation of the microphysiological environment in intestinal tissues. Furthermore, it can be customized on a large scale and enables high-throughput drug screening. The intestine-on-a-chip has widespread applications in various areas, including the development of intestinal disease models, as well as drug screening and testing. The technology behind the intestine-on-a-chip is based on a co-culture system of intestinal epithelial cells and other cell types. Such chips can not only construct three-dimensional structures such as intestinal microvilli and folds in combination with microengineering techniques, but also replicate the intricacies of the intestine. Microfluidic devices are employed to simulate biomechanical parameters like blood flow and physiological peristalsis within the intestinal tissues, thus giving such chips microphysiological characteristics akin to those of natural intestinal tissues and subjecting it to biomechanical regulation. This review provided an overview of the advances in intestine-on-a-chip technology, focusing on cell co-culture techniques, microfluidic technology, and microengineering techniques. Furthermore, the application of this technology in intestinal disease modeling, and drug screening and testing were summarized.
  • Hot Focus
    Research progress and prospects of pituitary organoids
    WANG Shengjie, JIANG Xiaoxia, GUO Qinghua
    2023, 47(8): 566-570. https://doi.org/10.7644/j.issn.1674-9960.2023.08.002
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    The pituitary gland is one of the most important endocrine organs in the human body and plays a key role in maintaining the homeostasis of the internal environment. Organoids, as a new type of in vitro research model, have been widely used in developmental biology, regenerative medicine, disease modeling, personalized precision medicine and other fields. In recent years, pituitary organoids derived from stem cells have provided a good experimental model for the research on pituitary diseases, and led to novel ideas for the research on treatment of pituitary diseases with regenerative medicine. This review focus on the advances in construction and applications of pituitary organoids.
    • Original articles
  • Original articles
    Regulation of ciliogenesis by ALS2
    HE Chunyu, ZHANG Biyu, JIAN Xiaoxiao, WANG Wen, ZHANG Yucheng, LI Huiyan, HU Huaibin
    2023, 47(8): 571-575. https://doi.org/10.7644/j.issn.1674-9960.2023.08.003
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    Objective To investigate the regulatory effect of amyotrophic lateral sclerosis protein 2（ALS2） on primary cilia and the possible mechanism. Methods The subcellular localization of ALS2 was detected with immunofluorescence. Subsequently, siRNA interference was used to knock down the content of ALS2 mRNA in RPE-1 cells specifically and the effect of reduced ALS2 protein content on primary cilia was studied. The protein level of ALS2 during ciliary assembly and disassembly was detected by Western blotting. A primary experimental system for ciliary disassembly was established to determine the effect of ALS2 protein deletion on the disassembly process of primary cilia. Finally, time-laps live-cell real time imaging was used to detect the effect of ALS2 on ciliary decapitation during disassembly of primary cilia. Results Immunofluorescence results showed that ALS2 was localized around the centrosome, which was the matrix of primary ciliogenesis. Knockdown of ALS2 in RPE-1 cells significantly increased the proportion of aberrant ciliogenesis. The results of dynamic detection of protein levels showed that the protein expression level of ALS2 gradually increased during the disassembly of primary cilia. Knocking down ALS significantly inhibited the cilia disassembly process. Live cell real time imaging showed that deletion of ALS2 protein inhibited ciliary decapitation during disassembly of primary cilia. Conclusion As a centrosome localization protein, ALS2 regulates the dynamic balance between primary ciliary assembly and disassembly by promoting the ciliary decapitation, and prevents the occurrence of ciliary diseases caused by abnormal growth of cilia.
  • Original articles
    Effect of BMP4 on expansion of mouse hematopoietic stem/progenitor cells in vitro
    LI Yunxing, ZHAO Jiahui, CHEN Keyi, ZHANG Bowen, LI Yunqiao, FAN Tao, HE Lijuan, PEI Xuetao, LI Yanhua
    2023, 47(8): 576-583. https://doi.org/10.7644/j.issn.1674-9960.2023.08.004
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    Objective To explore the effect of bone morphogenetic protein 4（BMP4） on the expansion of mouse hematopoietic stem/progenitor cells （HSPCs） in vitro. Methods c-Kit+ hematopoietic cells in mouse bone marrow were enriched with immune microbeads before being divided into the control group （without BMP4 addition） and the experimental group （supplemented with 40 ng/ml BMP4 every day）. After 4 days of culture,the viability of the cells was analyzed. The proportions and numbers of HSPCs in the cultured cells were calculated by flow cytometry. The ability of these cells to form hematopoietic colonies was investigated via hematopoietic colony-forming unit assay. Along with treatment with BMP4 signaling pathway inhibitor LDN193189 （500 nmol/L）,flow cytometry was used to detect the effect of blocking this signaling pathway on the proportions of HSPCs,while quantitative reverse transcription PCR（qRT-PCR）was used to analyze the expression levels of proteotoxic stress-related genes and apoptosis-related genes. Results After 4-day culture of c-Kit+ hematopoietic cells,the total cell numbers in the control group and the experimental group increased （3.20±0.33）fold and （5.00±0.61）fold,respectively. The results of flow cytometry analysis of the proportions of HSPCs in each group suggested that Lineage-Sca-1+c-Kit+（LSK） cells in the control group and the experimental group increased （2.62±0.09）fold and （5.67±0.11）fold,respectively. Hematopoietic colony-forming unit assay showed that（49.25±6.99） colonies were formed in the control group,compared with （69.95±3.40） colonies in the experimental group. The BMP4 signal pathway blocking experiment showed that the number of LSK cells was （3.16±0.43）×104 per well after 4-day culture supplemented with LDN193189 （500 nmol/L）,which was a（38.27±7.44）% decrease compared with the group without LDN193189. qRT-PCR detection results showed that compared with the control group,the expression levels of proteotoxic stress and apoptosis-related genes were decreased in the BMP4 treatment group. Conclusion BMP4 treatment can promote the expansion of mouse bone marrow hematopoietic cells in vitro,and improve the proliferation and colony formation ability of murine HSPC.
  • Original articles
    Compatibility of influenza virus RNP assessed using polymerase inducible reporter system
    YANG Wenguang, LI Wei, ZHAO Zhiyan, ZHANG Sen, CHEN Yuehong, HU Yi, JIANG Tao
    2023, 47(8): 584-589. https://doi.org/10.7644/j.issn.1674-9960.2023.08.005
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    Objective To establish a rapid monitoring and assessment model for mutated influenza A virus （IAV）, to evaluate the polymerase activity and subunit compatibility of the recombinant ribonucleoprotein （RNP） complex gene which is closely related to the replication level of IAV, and to monitor the new IAV. Methods Using the luciferase reporting system dependent on RNA polymerase activity of influenza virus, the polymerase activity of corresponding RNP heavy ligands among different subtypes of IAV and different strains of the same subtype was detected. Results The polymerase activity of virus RNP was not affected by the recombination of NP alone, but its activity was closely related to the source of the combination of PB1-PB2 and PB1-PA. The RNP polymerase activity of influenza virus PB1-PB2 from different host sources decreased significantly. Conclusion The luciferase reporting system, which depends on the activity of influenza virus RNA polymerase, can be used to assess the compatibility of different recombinant influenza virus RNPs.
  • Original articles
    Multiplex PCR and nanopore sequencing combined to detect gastrointestinal pathogens and its evaluation
    LIU Ran, YIN Xiaoyao, SHEN Yezhuang, ZHANG Yawei, HU Mandong, LI Weihua, ZHANG Xuemin, HE Kun, XIE Xianxing
    2023, 47(8): 590-595. https://doi.org/10.7644/j.issn.1674-9960.2023.08.006
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    Objective To establish a detection method that combines multiplex PCR and nanopore sequencing for high sensitivity and rapid detection of gastrointestinal pathogens on the scene. Methods Specific primers for gastrointestinal pathogens were designed before the specificity of the primers was verified and multiplex PCR amplification conditions were optimized in order to build a rapid library sequencing system and evaluate the sensitivity of detection. Results The established multiplex PCR detection system for twelve gastrointestinal pathogens could effectively amplify all the target sequences. The sensitivity of detection of the templates with mixed DNA from the twelve pathogens and the mixed RNA samples of three rotavirus A, B and C could reach 103 copies/ml when the concentration of each primer in the primer pool was 0.1 μmol/L. Conclusion With this method,gastrointestinal pathogens can be identified in a short time so that the efficiency of on-site detection of pathogens in public health emergencies can be improved. This method is expected to play an important role in disease control and prevention.
  • Original articles
    Effects of simulated microgravity on phenotype and genotype of Klebsiella pneumoniae
    WANG Xia, LIU Wenting, CHAI Zili, KANG Fuqiang, REN Hongguang, YUE Juejie, YE Xianwei, JIN Yuan
    2023, 47(8): 596-602. https://doi.org/10.7644/j.issn.1674-9960.2023.08.007
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    Objective To investigate the phenotypic and genotypic changes of Klebsiella pneumoniae under simulated microgravity（SMG）. Methods SMG on the ground was established by using a rotary cell culture system （RCCS）and K.pneumoniae was cultured under SMG and normal gravity（NG）. The growth rate, biofilm formation ability and drug sensitivity of K.pneumoniae were determined using the growth curve, crystal violet staining and paper diffusion method. The samples were sequenced by Illumina Miseq and analyzed for mutated genes. Results Compared with the NG group, the growth rate of K. pneumoniae under SMG slowed down, the degree of crystal violet staining deepened, and the sensitivity to various drugs changed significantly. Re-sequencing results showed that 19 genes were mutated. Two genes related to UDP-glucose and UDP-galactose were mutated under SMG, and the mutant genes under NG were more involved in signal transduction systems and biofilm formation pathways. Conclusion Under SMG, the growth rate of K.pneumoniae decreases, the ability of biofilm formation increases, the sensitivity of K.pneumoniae to various antimicrobial drugs changes, and two genes related to metabolism have been mutated.
  • Original articles
    Effects of different durations of tail-suspension on cardiac functions of mice under simulated weightlessness
    ZHOU Xuemei, FU Yesheng, ZHANG Lingqiang
    2023, 47(8): 603-608. https://doi.org/10.7644/j.issn.1674-9960.2023.08.008
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    Objective To investigate the changes of cardiac functions in mice under simulated weightlessness with different durations of tail-suspension. Methods Thirty mice were randomly divided into three groups： the control （CTRL） group, 40-day tail-suspension （TS-40 d） group and 70-day tail-suspension （TS-70 d） group. After tail suspension, the cardiac functions of mice under simulated weightlessness were evaluated by weighing, echocardiography, hematoxylin-eosin （HE） staining, Masson trichrome staining and real-time quantitative PCR （qPCR）. Results Compared with the CTRL group, body weight of the TS group mice was gained more slowly （P<0.01）. Echocardiography showed that the values of ejection fraction and fraction shortening of mice in the TS-40 d group were significantly decreased compared with the CTRL group （P<0.05）, and that these values were significantly lower than those of TS-70 d mice. The results of cardiac morphology and mass showed that the myocardial atrophy was caused by tail-suspension in the TS mice compared with the CTRL mice （P<0.0001）. HE and Masson trichrome staining showed that the collagen fibers in the heart of the TS group were blue, which was more obvious in the TS-70d group. The results of qPCR showed that the expressions of cardiac remodeling factor brain natriuretic peptide （BNP） and fibrosis marker matrix metalloproteinases-2（MMP2） were up-regulated in TS-40 d and TS-70 d groups, especially in the TS-70 d group. Conclusion TS-40 d mice show decreased cardiac function, cardiac atrophy and fibrosis while TS-70 d mice show adaptive recovery of cardiac function, but more severe cardiac atrophy and fibrosis.
  • Original articles
    EEG time-frequency analysis of impacts of mental fatigue induced by continuous operation on response inhibition
    ZHANG Chaoyue, DENG Haowei, XIE Fang, ZHAO Yongqi, FAN Ning, WANG Lubin
    2023, 47(8): 609-614. https://doi.org/10.7644/j.issn.1674-9960.2023.08.009
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    Objective To explore the effect of mental fatigue induced by continuous operation on response inhibition function based on the time domain and frequency domain characteristics of electroencephalogram（EEG） signals. Methods This study involved nineteen health male subjects. The 70-minute continuous cognitive operation was performed to induce mental fatigue. Go stimulus （response task） and Nogo stimulus （inhibition task） in the Go/Nogo task paradigm were used to evaluate behavioral changes before and after mental fatigue. Metrics of EEG analysis included the amplitude and latency of event-related potential N2 and P3,and brain oscillation rhythm of θ and β frequency band. Repeated measures ANOVA were used to analyze the EEG metrics before and after mental fatigue. Results ①Reaction time and correct rates of Go stimulus after mental fatigue didn't change significantly,but the correct rates of Nogo stimulus decreased significantly （P<0.01）. ②After mental fatigue,the N2-latency of Nogo stimulus on Cz electrode （P<0.001） and P3-latency of Go stimulus on Fz electrode （P<0.05） both prolonged significantly,while the P3-amplitude of both Go and Nogo stimuli on Fz and Cz electrodes decreased significantly（P<0.01）. ③On Fz electrode,mental fatigue caused β band power with 200-500 ms time window and 600-750 ms time window of Nogo stimulus to decrease significantly （P<0.05）. Conclusion The prolonged latency of N2 and decreased β band power are important neurophysiological features of impaired response inhibition function after mental fatigue.
  • Original articles
    Preparation methods and physicochemical properties of composite materials based on acellular bone matrix
    SUN Jiapei, DU jian, HUANG Xunwu, PENG Wei
    2023, 47(8): 615-620. https://doi.org/10.7644/j.issn.1674-9960.2023.08.010
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    Objective To improve the plasticity of artificial bone materials and observe their physical and chemical properties by physically mixing acellular bone matrix（ACBM） with calcium phosphate/calcium sulfate（CPC/CS）. Methods The cancellous bone of a healthy bovine femoral head was selected to prepare ACBM, which was used as the main body to fill calcium phosphate/calcium sulfate hemihydrate （CPC/CSH） to prepare a composite artificial bone scaffold. ACBM was set to account for 10%, 20%, and 30% of the total mass of the composite material respectively were measured before the mechanical properties （elastic modulus and compressive strength）. The physical and chemical properties of the experimental group with the best mechanical properties were analyzed with an X-ray diffractometer （XRD）, scanning electron microscope （SEM）, transmission electron microscope （TEM）, micro Kjeldahl nitrogen determination method, and liquid replacement method. The mechanical properties of the material with the best mechanical properties were compared with those of the allograft bone and composite CPC/CSH. Results ①The 20%ACBM composite bone material had the best mechanical properties among the composite materials with different proportions of ACBM. It could withstand a maximum of 923.81 N, a maximum deformation of 4.92 mm, an elastic modulus of （85.91±3.67） MPa, and a compressive strength of （3.34±0.15） MPa （P<0.001）. The XRD showed that this 20% ACBM composite artificial bone material could produce absorbable substances such as hydroxyapatite （HA）, CS, and calcium sulfate dihydrate （CSD） after solidification. The pore size of the material observed by scanning and transmission electron microscopy was （496.56±153.37） μm, the internal pores were interconnected, and the connectivity and defect filling degree were high. This material had an extremely low protein content. The porosity was（80.7±5.1）% as was measured with the liquid displacement method. ②The mechanical property test showed that the mechanical property of the 20% ACBM bone material was significantly better than that of the allograft bone group and CPC/CSH group （P<0.001）. Conclusion The 20% ACBM composite bone material has high mechanical properties and porosity,and is highly malleable.
  • Original articles
    Inpatients of military personnel in rehabilitation department of a military hospital in recent two years：a retrospective analysis
    ZHAO Chenyu, WANG Lanxiang, QI Changjie, QIAO Yijie, YANG Jihua, PU Rong, ZHANG Zishi
    2023, 47(8): 621-625. https://doi.org/10.7644/j.issn.1674-9960.2023.08.011
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    Objective To summarize the clinical data on inpatients of military personnel undergoing rehabilitation treatment in a rehabilitation department between 2020 and 2021,and explore the characteristics of training injuries among army personnel in order to providing reference for the prevention and treatment of training injuries and future research on rehabilitation. Methods By retrospective study, the medical records of military personnel hospitalized for treatment in the Department of Rehabilitation Medicine of our hospital over the past two years were collected before the information about gender, age, types of personnel, departments that made the first diagnosis, types of disease, injury sites, and surgery was analyzed. Results A total of 187 military personnel were enrolled into this study, 116（62%） of whom were hospitalized due to military training injuries and 71（38%）were hospitalized for other reasons. Most of these patients were males is aged 21 to 30. Bone and joint injuries were more common in both groups, and the lower limbs proved to be the most common injury sites in the military training group, accounting for 63.79%, while in the non-military training injury group, the trunk proved to be the most common injury site, accounting for 59.15%. In both groups, the orthopedic department was the first choice for diagnosis and treatment. Recipients of surgery accounted for 73.28% in the military training injury group, compared with 28.17% in the non-military training injury group. Conclusion In the past two years, most of the military personnel hospitalized in our department were injured during military training, and were young soldiers. The types of diseases were mostly bone and joint injuries, and the lower limbs proved to be the most common injury sites. Most of the patients underwent surgical treatment. Military training injuries, especially postoperative rehabilitation,deserve more attention.
    • Reviews
  • Reviews
    Research progress in mechanism and treatment of cytokine storm in sepsis
    SHI Yinghui, NIU Wenkai, YUAN Xin
    2023, 47(8): 626-630. https://doi.org/10.7644/j.issn.1674-9960.2023.08.012
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    Sepsis is a global health issue that poses a serious threat to human health. The high incidence and complex pathogenic mechanism of sepsis are the main causes of death in critically ill patients. Cytokine storm is a phenomenon characterized by a tremendous amount of cytokine production,which is manifested as the uncontrolled systemic inflammatory response caused by the overactive immune system and plays an important role in sepsis. This article reviews the research progress in the mechanism and treatment of cytokine storm in sepsis in order to provide new ideas and strategies for clinical practice.
  • Reviews
    Research progress in smallpox vaccines
    ZHANG Aihong
    2023, 47(8): 631-635. https://doi.org/10.7644/j.issn.1674-9960.2023.08.013
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    Smallpox has taken a heavy toll on human health in history. The World Health Organization （WHO） announced in 1980 that smallpox had been eradicated. Because of the likelihood of accidental leak of variola virus from leftover viral stocks and of the outbreak of a pandemic caused by monkeypox in recent years, there have been concerns about bioterrorism and public health risks caused by orthopox viruses. Vaccine is one of the efficient means to combat bioterrorism, contain epidemics and reduce public health risks. This paper reviews the progress in research on variola virus vaccines from the first generation （vaccinia virus strains） to the fourth generation （nucleic acid based vaccines） in terms of preparation technologies, incubation safety and efficacy.
  • Reviews
    Research progress in cognitive enhancement techniques for military personnel
    QIN Yilong, WEN Zhihong, CAO Xinsheng, WANG Chunchen
    2023, 47(8): 636-640. https://doi.org/10.7644/j.issn.1674-9960.2023.08.014
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    Given the profound influence of cognitive ability of military pilots on the skills of information acquisition, situational awareness and decision making in battlefield environments, cognitive enhancement targeting military pilots has become a hot topic in cognitive neuroscience. This paper elaborates the principles and effects of three intervention methods for cognitive enhancement of military pilots： cognitive training intervention, non-invasive brain stimulation technology, and cognitive enhancement drugs and supplements. Furthermore, prospective interventions will involve by big data modeling, artificial intelligence, and biomedical engineering so that cognitive enhancement research can become multi-modal, adaptive, and personalized. In the increasingly complicated and competitive battlefield environments, the progress in cross-disciplinary research on cognitive enhancement should be accelerated, which will help military pilots gain a competitive/edge on the battlefield in the future.
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