1.An experimental method for direct detection of lymphocyte γ-H2AX in mice peripheral blood and its application
Lei SHI ; Xing SHEN ; Ya DONG ; Qiaoyun ZHANG ; Hongling OU ; Xiujun SONG ; Yingying MA ; Xinru WANG
Chinese Journal of Radiological Medicine and Protection 2025;45(1):18-23
Objective:To develop a method of employing flow cytometry to directly detect the γ-H2AX expression levels in peripheral blood lymphocytes of mice through fixation and lysis and to evaluate the feasibility of applying this method to research on the radiation-related biological effects and the efficacy evaluation of radioprotective drugs.Methods:A total of 41 male C57BL/6J mice were used. First, 21 mice were randomly divided into 7 groups according to different radiation doses (0, 1, 2, 4, 6, 8, and 10 Gy) with 3 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation and immediately fixed with formaldehyde. Red blood cells (RBC) were lysed with Triton X-100, and γ-H2AX was labeled with specific antibodies. DRAQ5 dye was used to further exclude debris and anucleate cells. The mean fluorescence intensity of γ-H2AX in lymphocyte populations was directly analyzed by flow cytometry through forward and side scatter, and dose-effect curves after irradiation were established. Then, the other 20 mice were divided into radiation alone groups and radiation combined with WR-2721 administration groups at 4 and 6 Gy, respectively, with 5 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation to detect the average fluorescence intensity of γ-H2AX in lymphocytes, which was used to evaluate the degree of DNA damage in mice and the therapeutic effect of WR-2721.Results:The expression of γ-H2AX in peripheral blood lymphocytes of mice significantly increased with the increase of radiation doses, and reached a peak at 1-2 h and then decreased. The dose-effect relationship was significant ( R2 = 0.9914). At 24 h after 4 and 6 Gy irradiation, compared with the radiation alone groups, the average fluorescence intensity of γ-H2AX in the radiation combined with WR-2721 administration groups was lower (144.8 ± 8.0 and 109.5 ± 9.7, vs. 178.0 ± 18.5 and 136.6 ± 5.4), with statistically significant difference ( t = 3.78, 5.48, P < 0.05). The average fluorescence intensity of γ-H2AX at 24 h after irradiation was consistent with the lowest values of the three blood cell lines at 7 or 14 d after irradiation. Conclusions:The application of flow cytometry with a fixation/dissolution protocol to directly detect the mean fluorescence intensity of γ-H2AX in peripheral blood lymphocytes of mice has significant application value in radiation biology effect research, radiation protection drug screening, and efficacy evaluation.
2.The construction and application value of intelligent laboratory medicine
Miansheng WANG ; Yanle WANG ; Jianbo SUN ; Qiaoyun DAN ; Liang SHI ; Yuanbin LU
Chinese Journal of Laboratory Medicine 2025;48(2):201-206
Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.
3.Prevalence of functional gastrointestinal disorders among soldiers stationed on islands based on Rome Ⅳ diagnostic criteria
Muyun LIU ; Dan WANG ; Qiaoyun SHI ; Chang SUN
Journal of Navy Medicine 2025;46(2):122-126
Objective To investigate the prevalence of functional gastrointestinal disorders(FGID)in soldiers stationed on an island based on the Rome IV diagnostic criteria,so as to clarify the incidence and types of FGID and their correlations with the age and psychological status of soldiers.Methods The prevalence of FGID was investigated from September to November 2022 by FGID Questionnaire Survey Platform Management System based on the Rome IV diagnostic criteria.Results A total of 397 soldiers stationed on an island were enrolled in this study.The most common digestive symptoms were diarrhea,belching and constipation.A total of 147 people(37.0%)met the Rome Ⅳ diagnosis of FGID,and common diseases were functional diarrhea,epigastric pain syndrome and postprandial discomfort syndrome.Among the various diagnoses of FGID,the prevalence rates of globus hystericus,functional chest pain and belching syndrome showed statistically significant differences across different age groups(all P<0.001).Among the 324 subjects with no abnormal SDS scores,61.4%(199/324)did not meet the diagnosis of FGID.Among the 71 subjects with SDS scores ranging from 53 to 62,71.8%(51/71)did not meet the diagnosis of FGID.All subjects with SDS scores ranging from 63 to 72 met the diagnosis of FGID.Conclusion Functional diarrhea is the most common disease of FGID among soldiers stationed on the island.The overall prevalence of FGID is not related to the age of the subjects.Relevant drugs and psychological support treatment should be strengthened in the future allocation of health service resources on islands.
4.An experimental method for direct detection of lymphocyte γ-H2AX in mice peripheral blood and its application
Lei SHI ; Xing SHEN ; Ya DONG ; Qiaoyun ZHANG ; Hongling OU ; Xiujun SONG ; Yingying MA ; Xinru WANG
Chinese Journal of Radiological Medicine and Protection 2025;45(1):18-23
Objective:To develop a method of employing flow cytometry to directly detect the γ-H2AX expression levels in peripheral blood lymphocytes of mice through fixation and lysis and to evaluate the feasibility of applying this method to research on the radiation-related biological effects and the efficacy evaluation of radioprotective drugs.Methods:A total of 41 male C57BL/6J mice were used. First, 21 mice were randomly divided into 7 groups according to different radiation doses (0, 1, 2, 4, 6, 8, and 10 Gy) with 3 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation and immediately fixed with formaldehyde. Red blood cells (RBC) were lysed with Triton X-100, and γ-H2AX was labeled with specific antibodies. DRAQ5 dye was used to further exclude debris and anucleate cells. The mean fluorescence intensity of γ-H2AX in lymphocyte populations was directly analyzed by flow cytometry through forward and side scatter, and dose-effect curves after irradiation were established. Then, the other 20 mice were divided into radiation alone groups and radiation combined with WR-2721 administration groups at 4 and 6 Gy, respectively, with 5 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation to detect the average fluorescence intensity of γ-H2AX in lymphocytes, which was used to evaluate the degree of DNA damage in mice and the therapeutic effect of WR-2721.Results:The expression of γ-H2AX in peripheral blood lymphocytes of mice significantly increased with the increase of radiation doses, and reached a peak at 1-2 h and then decreased. The dose-effect relationship was significant ( R2 = 0.9914). At 24 h after 4 and 6 Gy irradiation, compared with the radiation alone groups, the average fluorescence intensity of γ-H2AX in the radiation combined with WR-2721 administration groups was lower (144.8 ± 8.0 and 109.5 ± 9.7, vs. 178.0 ± 18.5 and 136.6 ± 5.4), with statistically significant difference ( t = 3.78, 5.48, P < 0.05). The average fluorescence intensity of γ-H2AX at 24 h after irradiation was consistent with the lowest values of the three blood cell lines at 7 or 14 d after irradiation. Conclusions:The application of flow cytometry with a fixation/dissolution protocol to directly detect the mean fluorescence intensity of γ-H2AX in peripheral blood lymphocytes of mice has significant application value in radiation biology effect research, radiation protection drug screening, and efficacy evaluation.
5.The construction and application value of intelligent laboratory medicine
Miansheng WANG ; Yanle WANG ; Jianbo SUN ; Qiaoyun DAN ; Liang SHI ; Yuanbin LU
Chinese Journal of Laboratory Medicine 2025;48(2):201-206
Objective:To investigate the application value of intelligent laboratory construction in the laboratory.Methods:All samples sent to the Department of Clinical Laboratory from all the clinical departments at the Eighth Affiliated Hospital of Sun Yat-Sen University from January 2018 to June 2024 were collected. Full-process intelligentization building was achieved by the use of logistics transmission system, automatic sampling system, automatic quality control system, automatic audit system, intelligent monitoring system, critical value monitoring system, and intelligent DxlabReport operation management system, et al. The normal distribution data were analyzed by two independent samples t-test, and the skew distribution data were analyzed by Mann-Whitney U test. Results:After the implementation of the pneumatic logistics transmission system, the turn-around time (TAT) from sample transmission to receiving decreased significantly from 51.5 (37.7, 73.0) min to 18.1 (14.4, 31.0) min ( U=0, P<0.001). After adopting the automatic sampling system, the TAT from receiving to sampling decreased from 41.0 (38.0, 45.0) min to 10.0 (5.0, 11.0) min ( U=0, P<0.001). The introduction of automatic quality control advanced the median time for the first batch of samples entering the biochemical testing pipeline from 9∶03 to 8∶27 and increased the reporting rate within 3 hours from 27.71%±1.39% to 36.90%±2.30% ( t=3.423, P<0.001). The intelligent monitoring system enabled module positioning monitoring, sample turn-around time reminder, instrument load rate monitoring, remaining reagent monitoring, and patient-based real-time quality control, resulting in improved instrument running efficiency and result accuracy. Followed by the introduction of the automatic audit function, the overall pass rate was 28.19%(10 006/35 500), including 37.17% (7 738/20 818) for biochemical reports, 31.57% (1 251/3 963) for chemiluminescence reports, and 9.49% (1 017/10 719) for biochemical immunity reports. The laboratory TAT decreased from (207.3±6.0) min to (169.8±5.9) min ( t=4.426, P<0.001). After the implementation of the critical value monitoring system, the timely reporting rate reached 99.52% (99.32%, 99.89%). After using quality digital management, outpatient biochemical immunity process was optimized to a decrease in laboratory TAT from 222 (201, 233) min to 145 (119, 195) min ( U=0, P=0.004), while the pass rate increased from 86.88% (85.91%, 87.81%) to 96.32% (95.86%, 96.96%) ( U=0, P=0.004). Conclusion:The establishment of an intelligent laboratory can optimize workflow, significantly improve the work efficiency and accuracy of sample processing, and minimize error.
6.Advances in research and application of ionizing radiation biomarkers
Hongling OU ; Wenchao AI ; Yan WANG ; Yingying MA ; Lei SHI ; Qiaoyun ZHANG ; Xiujun SONG ; Xinru WANG
Chinese Journal of Pharmacology and Toxicology 2024;38(1):70-78
Exposure to ionizing radiation intervenes in genomic stability and gene expression,resulting in the disruption of normal metabolic processes in cells and organs by causing complex biolog-ical responses.Altered genomic variations,gene expression and metabolite concentrations in blood or tissue samples reflect systemic radiation damage.With the application of new techniques and exten-sive study on the mechanisms for ionizing radiation damage,related indicators such as chromosomal variation,gene expression,lipid and metabolism are being recognized and promise to be the markers for early diagnosis and prognosis of radiation exposure.Therefore,this article reviews recent progress in and potential applications of biomarkers related to ionizing radiation injury.
7.The mediating role of psychological capital between occupational stress and suicidal ideation
Qianqian GAO ; Yuxiao WANG ; Jian SHI ; Xiaojun LIANG ; Yanyan SUN ; Qiaoyun ZHANG ; Pei LIU
Chinese Journal of Preventive Medicine 2020;54(11):1207-1212
Objective:To analyze the mediating effect of psychological capital (PsyCap) between occupational stress and suicidal ideation.Methods:In September 2017, stratified-cluster sampling method was adopted to select employees of an electronic manufacturing service industry as respondents for a questionnaire survey in Jiangsu province. The demographic characteristics of employees were collected. A total of 2 465 employees were investigated, and 2 216 valid questionnaires were collected. The level of occupational stress and PsyCap was assessed by the Effort-Reward Imbalance Scale(ERI)and PsyCap Questionnaire.Results:The age of 2 216 employees was (28.70±5.26) years old, including 1 230 males (55.5%). The scores of occupational stress on effort, reward and over-commitment were (3.08±0.63), (3.73±0.60) and (2.51±0.68), respectively. The average ERI index (ratio of effort to return) was (0.88±0.39). The total score of PsyCap was (86.62±15.23). The detection rate of suicidal ideation within two weeks was 13.9% (309/2 216). The average score of effort and over-commitment and the ERI index were positively correlated with the number of suicidal ideation detected within two weeks, with correlation coefficients about 0.081, 0.141 and 0.164, respectively. The average score of reward and total score of PsyCap were negatively correlated with the number of suicidal ideation detected within two weeks, with correlation coefficients about -0.206 and -0.228, respectively. The mediating effect results showed that PsyCap had mediating effect on the effort, reward, over-commitment and suicidal ideation of occupational stress respectively, with mediating effect values of -0.087 (95% CI: -0.018,-0.019), -0.299 (95% CI: -0.428,-0.200) and 0.096 (95% CI: 0.0394, -0.170). The mediating effect percentages were 9.43%, 17.92%, and 9.54%, respectively. Conclusion:PsyCap has a mediating effect between effort, reward, over-commitment of occupational stress and suicidal ideation.
8.The mediating role of psychological capital between occupational stress and suicidal ideation
Qianqian GAO ; Yuxiao WANG ; Jian SHI ; Xiaojun LIANG ; Yanyan SUN ; Qiaoyun ZHANG ; Pei LIU
Chinese Journal of Preventive Medicine 2020;54(11):1207-1212
Objective:To analyze the mediating effect of psychological capital (PsyCap) between occupational stress and suicidal ideation.Methods:In September 2017, stratified-cluster sampling method was adopted to select employees of an electronic manufacturing service industry as respondents for a questionnaire survey in Jiangsu province. The demographic characteristics of employees were collected. A total of 2 465 employees were investigated, and 2 216 valid questionnaires were collected. The level of occupational stress and PsyCap was assessed by the Effort-Reward Imbalance Scale(ERI)and PsyCap Questionnaire.Results:The age of 2 216 employees was (28.70±5.26) years old, including 1 230 males (55.5%). The scores of occupational stress on effort, reward and over-commitment were (3.08±0.63), (3.73±0.60) and (2.51±0.68), respectively. The average ERI index (ratio of effort to return) was (0.88±0.39). The total score of PsyCap was (86.62±15.23). The detection rate of suicidal ideation within two weeks was 13.9% (309/2 216). The average score of effort and over-commitment and the ERI index were positively correlated with the number of suicidal ideation detected within two weeks, with correlation coefficients about 0.081, 0.141 and 0.164, respectively. The average score of reward and total score of PsyCap were negatively correlated with the number of suicidal ideation detected within two weeks, with correlation coefficients about -0.206 and -0.228, respectively. The mediating effect results showed that PsyCap had mediating effect on the effort, reward, over-commitment and suicidal ideation of occupational stress respectively, with mediating effect values of -0.087 (95% CI: -0.018,-0.019), -0.299 (95% CI: -0.428,-0.200) and 0.096 (95% CI: 0.0394, -0.170). The mediating effect percentages were 9.43%, 17.92%, and 9.54%, respectively. Conclusion:PsyCap has a mediating effect between effort, reward, over-commitment of occupational stress and suicidal ideation.
9.Clinical research on the modified nasopharyngeal airway tube combined with intubation soft lens in endotracheal intubation
Fusheng XU ; Meihong ZHUANG ; Sha DENG ; Zhibo SHI ; Renfang YOU ; Qiaoyun LI
Journal of Navy Medicine 2017;38(2):145-148
Objective To observe clinical effects of the modified nasopharyngeal airway tube combined with intubation soft lens in endotracheal intubation.Methods Two hundred patients who were to receive surgery were chosen as research subjects.With the knowledge and consent of the patients and following the signing of the letter of agreement,the patients were divided into 4 groups:group A,B,C and D,each consisting of 50 patients.For the patients of group A and B,endotracheal intubation was performed by high-qualified anesthesiologists (with the certificate of medical practitioner for over 5 years),while for the patients of group C and D,intubation was done by low-qualified ones (with the certificate of medical practitioner for less than 5 years).For the patients of group A and C,nasopharyngeal airway tube was placed through the approach of side nasal cavity following anesthesia induction,and then soft lens was used to complete endotracheal intubation via nasopharyngeal airway.For the patients of group B and D,soft lens intubation was performed directly through nasal cavity following anesthesia induction.Medical data,such as intubation time,number of intubation,first time of glottis exposure,cases of intubation failure,and adverse reactions or complications induced by intubation were recorded for further analysis.Results The intubation time of group A was shorter than that of group 13,and the time of group C was shorter than that of group D,and statistical significance could be seen,when comparisons were made between them(P < 0.05).The success rate at the first try of intubation for group A and C were respectively 84% and 82%,and the failure rate was all 2%,while for group B and D the success rate of intubation were respectively 50% and 30%,and failure rates were respectively 10% and 20%.Statistical significance could be noticed,when comparisons were made between the groups,i.e.between group A and B,between group C and D and also between group B and D (P < 0.05).Conclusion Modified nasopharyngeal airway tube combined with intubation soft lens could obviously shorten endotracheal intubation time and increase the success rate of intubation.No significant differences could be noted in intubation time and failure rate,when the use of modified nasopharyngeal airway tube was combined with intubation soft lens by high-qualified and low-qualified anesthesiologists.
10.In vitro expression and analysis of the 826 human G protein-coupled receptors.
Xuechen LV ; Junlin LIU ; Qiaoyun SHI ; Qiwen TAN ; Dong WU ; John J SKINNER ; Angela L WALKER ; Lixia ZHAO ; Xiangxiang GU ; Na CHEN ; Lu XUE ; Pei SI ; Lu ZHANG ; Zeshi WANG ; Vsevolod KATRITCH ; Zhi-Jie LIU ; Raymond C STEVENS
Protein & Cell 2016;7(5):325-337
G protein-coupled receptors (GPCRs) are involved in all human physiological systems where they are responsible for transducing extracellular signals into cells. GPCRs signal in response to a diverse array of stimuli including light, hormones, and lipids, where these signals affect downstream cascades to impact both health and disease states. Yet, despite their importance as therapeutic targets, detailed molecular structures of only 30 GPCRs have been determined to date. A key challenge to their structure determination is adequate protein expression. Here we report the quantification of protein expression in an insect cell expression system for all 826 human GPCRs using two different fusion constructs. Expression characteristics are analyzed in aggregate and among each of the five distinct subfamilies. These data can be used to identify trends related to GPCR expression between different fusion constructs and between different GPCR families, and to prioritize lead candidates for future structure determination feasibility.
Animals
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Computational Biology
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Crystallography, X-Ray
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Gene Expression
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Humans
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Plasmids
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genetics
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metabolism
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Protein Domains
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Receptors, Adrenergic, beta-1
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Receptors, G-Protein-Coupled
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classification
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genetics
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metabolism
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Receptors, Odorant
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metabolism
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Receptors, Purinergic P1
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genetics
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metabolism
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Sf9 Cells
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Spodoptera

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