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    A novel model for drug discovery based on proteolysis-targeting chimeras （PROTAC）
    LI Bing-kun, YANG Ning, LI Peng-yun, ZHENG Zhi-bing
    2023, 47(1): 1-12. https://doi.org/10.7644/j.issn.1674-9960.2023.01.001
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Proteolysis-targeting chimeras （PROTAC） is a bifunctional molecule that targets protein of interest （POI） for degradation by co-opting the ubiquitin-proteasome system （UPS）. In recent years, PROTAC technology has been widely explored as a novel therapeutic strategy for cancer, autoimmune diseases, neurodegenerative diseases and viral infection, which has become a hot spot and priority for drug development. This review is intended to outline the important milestones and the key research progress in the field of PROTAC, offer recommendations on the design of PROTAC,and describe the challenges to and prospects of targeted degradation by PROTAC.
    • Original articles
  • Original articles
    Progress in and strategic analysis of research and development of vaccines in the U.S. Armed Forces
    ZU Mian, WANG Lei
    2023, 47(1): 13-19. https://doi.org/10.7644/j.issn.1674-9960.2023.01.002
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Vaccines are considered a powerful weapon against major epidemics. As one of the most successful and cost-effective public health interventions, vaccination plays an important role in controlling widespread infections and maintaining military strength. The US military medical institutions have been committed to vaccine research and development （R&D） for many years. They have developed numerous effective vaccines against bacterial and viral infections, which play a crucial role in global deployment of the U.S. Armed Forces. The main spheres, priorities and characteristics of related research in these institutions are of significant referential value. This paper outlines the main research institutions engaged in vaccine research and the representative vaccines currently in preclinical and clinical trials, and summarizes the main strategies of the U.S military to promote R&D of vaccines, which is expected to provide data for R&D of vaccines in China′s armed forces.
  • Original articles
    Protective effect and mechanism of TGF-β3 on lung epithelial cell injury induced by radiation
    YAN Cheng-ming, YANG Zhi-hua, WANG Yi-long, GENG Shuang, LIU Ben-bo, WANG Zhi-xin, LI Qian, WANG Mei-yu, GUO Hao-xin, ZHU Mao-xiang
    2023, 47(1): 20-25. https://doi.org/10.7644/j.issn.1674-9960.2023.01.003
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    Objective To explore the role and possible mechanism of transforming growth factor-β3 （TGF-β3） in radiation-induced lung epithelial cell injury, and to provide data for the prevention and treatment of radiation-caused lung injury. Methods The type Ⅱ transforming growth factor -β receptor （TGFBR2） siRNA was designed, synthesized and screened for cell TGFBR2 interference. Human bronchial epithelial cells （Beas-2B cells） and their TGFBR2 interference cells were randomly divided into the control group, 5 ng/ml TGF-β3 acting group （TGF-β3 group）, 6 Gy 60Co-γ-ray irradiation group （6 Gy group）, and 6 Gy 60Co-γ-ray irradiation+5 ng/ml TGF-β3 action group （6 Gy+TGF-β3 group）. The expressions of collagens and TGFBR2 in each group were detected by real-time fluorescence quantitative PCR（RT-qPCR）, and the expression of smooth muscle actin （α-SMA）, a marker of epithelial-mesenchymal transition（EMT）, was detected by RT-qPCR and Western blotting. Cell apoptosis and cell cycle were analyzed by flow cytometry. Results （1） TGF-β3 could significantly inhibit the expressions of collagens induced by radiation and reduce the abnormally high expressions of α-SMA gene and protein, which was a marker of EMT induced by radiation, suggesting that TGF-β3 could effectively protect against lung cell fibrosis induced by radiation. （2） TGF-β3 could inhibit abnormally high expressions of TGFBR2 induced by radiation, and had significant protective effect on cell apoptosis and G2/M phase arrest induced by radiation.（3） The protective effect of TGF-β3 on radiation-induced apoptosis and G2/M cell cycle arrest was lost after TGFBR2 interference, suggesting that TGF-β3 exerted its inhibitory effect on radiation-induced lung epithelial cell injury through TGFBR2. Conclusion TGF-β3 can protect radiation-damaged lung epithelial cells through TGFBR2.
  • Original articles
    Screening of discrepant genes in lung tissues of pulmonary fibrosis susceptible/resistant mice by telomere and telomerase PCR microarray
    JIA Zhao-qian, ZHANG Xiao, XING Yuan, YAN Hua, ZHAO Xin-yue, LI Xiao-yu, WANG Yong-yi, YE Yu-meng, HAO Yan-hui, ZUO Hong-yan, LI Yang
    2023, 47(1): 26-32. https://doi.org/10.7644/j.issn.1674-9960.2023.01.004
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    Objective To establish a model of pulmonary fibrosis-prone and fibrosis-resistant mice and screen the differential genes in lung tissues before and after irradiation by telomere and telomerase real-time fluorescence quantitative PCR（RT-qPCR） microarray. Methods The chests of C57BL/6J and C3H/HeN mice were irradiated with 20 Gy 60Co gamma rays. The differences in radiation-induced pulmonary fibrosis（RIPF） progression between them were studied via HE staining, sirius red staining, hydroxyproline content, TGF- β and CTGF detection. Telomere and telomerase RT-qPCR microarray was used to compare the expressions of telomere and telomerase related genes in their lung tissues before irradiation and 3 months after irradiation. The differential genes were verified by RT-qPCR. Results The fibrotic lesions of C57BL/6J mice were more obvious than those of C3H/HeN mice 3 months after irradiation. The content of hydroxyproline in lung tissues of C57BL/6J mice was significantly higher than before irradiation and in C3H/HeN mice（P<0.05）, and much more collagen typeⅠwas deposited in the alveolar wall of C57BL/6J mice than in that of C3H/HeN mice. The contents of TGF-β and CTGF in lung tissues of C57BL/6J mice were significantly higher than in those of C3H/HeN mice（P<0.05）. There were 48 differential genes in telomere and telomerase related pathways in lung tissues of control C57BL/6J and C3H/HeN mice and 32 differential genes were found in the lung tissues of the two types of mice 3 months after irradiation. Moreover, 8 differential genes were discovered in the lung tissues of C57BL/6J mice 3 months after irradiation and 4 discrepant genes were detected in the lung tissues of C3H/HeN mice after irradiation. The differential gene Tnks2 was verified. The expression of Tnks2 in lung tissues of C3H/HeN mice 3 months after irradiation was higher than that of C57BL/6J mice and before irradiation（P<0.05）. Conclusion Differential genes in telomere and telomerase related pathways in lung tissues of fibrosis-susceptible and fibrosis-resistant mice are found before and after 60Co gamma rays irradiation.
  • Original articles
    Effect of purification process on oxygen carrying/releasing capacity of bovine hemoglobin
    QIN Dong, LI Wei-dan, SHENG Ming, GAO Dao-yuan, CHU Zong-tang, ZHOU Hong, CHENG Huan, WANG Ying
    2023, 47(1): 33-40. https://doi.org/10.7644/j.issn.1674-9960.2023.01.005
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Objective To study the effects of different experimental conditions on the oxygen-carrying/oxygen-releasing capacity of bovine hemoglobin in the purification process, and to screen a method for obtaining stable hemoglobin with oxygen-carrying/oxygen-releasing capacity in order to recommend a process for the preparation of raw materials for hemoglobin-based oxygen carriers（HBOCs）. Methods The low-osmotic hemolysis method, microfiltration, anion exchange chromatography and ultrafiltration were used to isolate and purify bovine hemoglobin from fresh bovine blood. In the process of purification, bovine hemoglobin was analyzed with the red blood cell oxygen carrying/releasing function evaluation device, and the concentration, purity and recovery of methemoglobin（MetHb） in the purified samples were detected so as to screen the purification conditions. Results The optimal purification conditions were determined as follows： PB solution was used for hemolysis, PVDF membrane was used for microfiltration, chromatography conditions were Buffer A （20 mmol/L Tris-HCl, pH 8.0） and Buffer B （20 mmol/L Tris-HCl+0.15 mol/L NaCl, pH 8.0）, and PBS buffer was used for ultrafiltration. This method had little effect on the oxygen carrying/ releasing capacity of bovine hemoglobin. The P50 （CV） of purified bovine hemoglobin was （28.69±0.11） mmHg（0.40%）, Hill coefficient was （2.49±0.04）（1.52%）, the hemoglobin protein purity exceeded 99%, and the MetHb concentration was lower than 1%. Conclusion Different experimental conditions in the purification process may have different effects on the oxygen carrying/releasing capacity of bovine hemoglobin. This study can contribute to screening the optimal process parameters.
  • Original articles
    Identification and immunological study of recombinant adenovirus with Gn of severe fever with thrombocytopenia syndrome virus
    LIU Wen-kai, TIAN Li, LIU Le-le, SHI Chen-juan, YANG Pan, HAN Yue, ZHAO Zhong-xin, XIA Xian-zhu, ZHENG Xue-xing, ZHENG Wen-wen
    2023, 47(1): 41-46. https://doi.org/10.7644/j.issn.1674-9960.2023.01.006
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    Objective To obtain a recombinant human adenovirus type 5 vector （Ad5） vaccine expressing the glycoprotein N （Gn） of severe fever with thrombocytopenia syndrome virus （SFTSV）, and to identify its biological characteristics and production of neutralizing antibodies. Methods The AdEasy adenovirus packaging system was used to construct a pAD-Amp-Gn expression vector and transfect it into HEK-293 cells to obtain recombinant adenovirus Ad5-Gn. After infecting the cells with Ad5-Gn, the expression of SFTSV Gn was identified by immunofluorescence and Western blotting. BALB/c mice were injected with Ad5-Gn to observe the behavior and detect the level of specific neutralizing antibodies, and to evaluate the safety and immunogenicity of Ad5-Gn. Results Immunofluorescence and Western blotting showed that SFTSV Gn expression could be detected in Ad5-Gn infected cells, and the size was as big as expected. Ad5-Gn did not affect the growth of mice, and the average titer of neutralizing antibodies produced after 4 weeks of infection was 1∶341. Conclusion An Ad5-Gn vaccine candidate expressing SFTSV Gn has been obtained, which has low virulence and can induce high levels of neutralizing antibodies in mice.
  • Original articles
    EDⅢ K307 is an important site for West Nile virus infection
    LU Xing, CHEN Nan, WANG Xun, CHEN Guo-jiang, WANG Jing, QIAO Chun-xia, LI Xin-ying, LIU Cheng-hua, SHEN Bei-fen, FENG Jian-nan, XIAO He
    2023, 47(1): 47-50. https://doi.org/10.7644/j.issn.1674-9960.2023.01.007
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    Objective To investigate the effect of the E protein DⅢ K307 site on the packaging and invasion of West Nile virus. Methods 293T cells were cotransfected with pCDNA3.1-CME K307A and prWNV-RLuc. The supernatant of cell culture was collected 72 h later. To find out whether the pseudovirus was packaged properly, the morphology of pseudotype virions in the supernatant was observed by electron microscopy, and the NS1 fragment sequence of viral genome was amplified by PCR. Then the infectivity of the mutated pseudovirus to target cells was determined by infecting BHK21 cells with the supernatant. Results Viral particles were uniform in the cell culture supernatant, round and enveloped, with a diameter of about 40-50 mm. Genomic NS1 specific bands were observed by PCR amplification. All the above results confirmed the successful packaging of mutated pseudoviral particles. However, infection experiments showed that the mutated pseudovirions at the EDⅢ K307 site lost the ability to infect target cells compared with the wild type pseudovirions. Conclusion West Nile virus EDⅢ K307 site is probably involved in infection, possible the key site for West Nile virus to bind to receptors, with may be an important target for anti- West Nile virus drugs.
  • Original articles
    Deviation factors for determination of therapeutic drug concentrations via chemiluminescent particle immunoassay
    CHEN Yan, YANG Fan, JIANG Nan, WANG Tian-lin
    2023, 47(1): 51-55. https://doi.org/10.7644/j.issn.1674-9960.2023.01.008
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    Objective To analyze the influencing factors of deviation during determination of therapeutic drug concentrations using chemiluminescent particle immunoassay （CMIA）. Methods The concentrations of cyclosporine A, tacrolimus, valproic acid, vancomycin and digoxin were measured in clinical samples and quality controls using CMIA respectively. Results After reagent calibration, the quality controls of cyclosporine A, tacrolimus, valproic acid, vancomycin and digoxin "hit the targets" accurately. The concentrations of cyclosporine A and tacrolimus decreased by 10%-16% when the reagents were used for 2 months. The concentrations of valproic acid, vancomycin and digoxin decreased by 15%-20% when the reagents were used for 3 months. Blood concentration samples of cyclosporine A had to be discarded after being stored for more than 7 days at 2-8℃, and digoxin samples should not be stored for more than 3 days at room temperature or 2-8℃. Pre-treated cyclosporine A and tacrolimus samples should be tested as soon as possible within 2 h. Conclusion Accurate calibration and immediate use of reagents plus proper storage, quick preprocessing and testing of samples can effectively reduce deviation of drug concentrations during determination with CMIA.
  • Original articles
    Ultrasonic diagnostic criteria for nephroptosis in Air Force medical recruitment
    ZHAO Guo-zheng, LIU Shu-ping, LI Li, AN Ping, SUN An, ZHU Liang, CHEN Xiao, GONG Feng-ling, SUN Bin
    2023, 47(1): 56-59. https://doi.org/10.7644/j.issn.1674-9960.2023.01.009
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    Objective To explore the ultrasound diagnostic criteria for nephroptosis in medical selection of military pilot cadets. Methods The proportion and mobility of kidneys under the costal arch were measured in 338 candidates recruited for Air Force medical selection in 2021. Results ①The proportion of the kidney under the costal arch of the left kidney in the standing position was smaller than that of the right kidney. ②The left kidney had greater mobility than the right one. The greater the proportion of the kidney under the costal arch in the upright position, the greater the mobility of the kidney. ③The upper limits of 95% and 99% reference ranges of bilateral kidney mobility were 5.5 cm and 7.3 cm for the right kidney and 5.7 cm and 7.0 cm for the left kidney, respectively. ④The ROC curve was drawn based on the upper limit of 95% reference range. The critical value, sensitivity, specificity and area under the curve were 0.66, 100.0%, 82.7% and 0.937 for the right kidney and 0.46, 93.8%, 69.3% and 0.872 for the left kidney. Conclusion It is recommended that according to the ultrasonic diagnostic criteria for nephroptosis, candidates who have any of the following conditions be considered unqualified： ①The whole kidney, right or left, is located under the costal arch. ②The mobility of the right kidney is greater than or equal to 7.3 cm or the mobility of the left kidney is greater than or equal to 7.0 cm. ③The right kidney exceeds 2/3 of the renal length under the costal arch and the mobility is greater than or equal to 5.5 cm, or the left kidney exceeds 1/2 of the renal length under the costal arch and the mobility is greater than or equal to 5.7 cm.
  • Original articles
    Ophthalmic strategies against COVID-19 epidemic at an African peacekeeping hospital
    WEI Jie, ZHAO Yong-Qiang, ZHAO Hui-jing, JIANG Hua, FU Qiang
    2023, 47(1): 60-64. https://doi.org/10.7644/j.issn.1674-9960.2023.01.010
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Objective To summarize the prevention and control measures taken against the coronavirus diesease 2019（COVID-19） pandemic in an ophthalmology department of a peacekeeping hospital in Africa, and to provide reference for subsequent peacekeeping medical teams in Africa to deal with such public health events. Methods Based on the actual conditions of the peacekeeping hospital, ophthalmological strategies were summarized in terms of clinic adjustment, E-mail appointment, graded protection, medical procedures and patient management. Results A total of 86 ophthalmic patients （101 times） were admitted to the peacekeeping hospital during one-year mission, including 21 emergency cases. Seventy-nine slit-lamp examinations,8 fundus examinations using anterior mirrors, 10 ophthalmic operations, 4 conjunctival sac irrigations and 3 ocular B ultrasound examinations were performed respectively. Most procedures were carried out under secondary protection, while conjunctival sac irrigation and debridement and suturing were performed under tertiary protection. All patients were well treated, and no novel coronavirus infections were detected in any of medical staff at the end of the mission. Conclusion The ophthalmic prevention and control strategies at this peacekeeping hospital in Africa against COVID-19 epidemic are indeed effective, and can provide reference for better ophthalmic health support in overseas peacekeeping operations.
    • Reviews
  • Reviews
    Adverse effects of military operational stress on muscle function and strategies for protection
    JIN Hong, HE Jun, ZHANG Li, JIA Li, GUO Jia-bin
    2023, 47(1): 65-70. https://doi.org/10.7644/j.issn.1674-9960.2023.01.011
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Military personnel may be exposed to external stressors, such as large energy deficits, sleep deprivation, cognitive demands, and environmental extremes during training and military operations that combine to degrade muscle protein. Exposure to these factors results in a hypogonadal state that may contribute to observed decrements in muscle mass. In this review, lessons learned from studying severe clinical stressed states and the interventions designed to mitigate the loss of muscle protein are discussed in the context of training and military operational stress. Restoration of the anabolic hormonal status in stressed states may be necessary to restore the anabolic in?uence derived from dietary protein on muscle. Based on clinical experience, restoration of the normal testosterone status during sustained periods of operational stress may be advantageous. Furthermore, an optimal protein intake, essential amino acid delivery in particular, and resistance training may be essential to the restoration of anabolic response during the stress state. Interventions that are intended to improve the protein balance of muscle networks may have a positive impact on the effectiveness of military personnel under difficult combat conditions.
  • Reviews
    Oxidative activation of ATM and multiple roles of ATM in oxidative stress
    LIU Xiao-lan, YAN Xin-long, ZHENG Xiao-fei
    2023, 47(1): 71-74. https://doi.org/10.7644/j.issn.1674-9960.2023.01.012
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    Ataxia-telangiectasia mutated （ATM） is a protein encoded by ATM gene. ATM as an important factor regulating oxidative stress response can maintain redox homeostasis in vivo. This review discusses the mechanism of ATM activation by oxidative stress,the role of ATM activation in alteration of cell redox homeostasis,cell metabolism and autophagy,and its relationship with diseases. It provides a new idea for future study on the biological function of ATM and its application to the treatment of related diseases.
  • Reviews
    Advances in non-invasive drug delivery across the blood-brain barrier
    YOU Li-li, FU Wen-liang, ZHANG Chao, XU Dong-gang
    2023, 47(1): 75-80. https://doi.org/10.7644/j.issn.1674-9960.2023.01.013
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    The blood-brain barrier （BBB） is the main bottleneck for drug treatment of the central nervous system（CNS） diseases. The BBB protects the brain from such harmful substances as bacteria and viruses, but it also constrains the entry of many drugs into brain tissue to exert its efficacy. Non-invasive drug delivery technologies targeting the brain have been developing rapidly, with a great potential for various applications. This article reviews the characteristics and applications of various non-invasive drug delivery technologies targeting the brain based on cell-penetrating peptides, cell carriers, specific receptors and nanomaterials in order to provide data for drug therapies of central nervous diseases.
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