1.Role of the iNOS/IRS1/AKT/GSK-3β signaling pathway in chronic intermittent hypoxia-induced insulin resistance
Meina Jin ; Xueli Zhou ; Haibo Li ; Wei Bai ; Chuxuan Jia ; Li Gao ; Lijue Ren ; Qingyu Chen ; Rui Wang ; Hua Li ; Cuiying Wei
Acta Universitatis Medicinalis Anhui 2025;60(2):210-217
Objective :
To pathological changes and inducible nitric oxide synthase(iNOS), phosphorylated insulin receptor substrate 1 serine 307(p-IRS1ser 307), phosphorylated protein kinase B serine 473(p-AKTser 473), glycogen synthase kinase-3β(GSK-3β), and gluconeogenic synthase(GS) proteins were observed in the liver of rats under the condition of chronic intermittent hypoxia-replicated oxygen in control. And to explore the role of iNOS/IRS1/AKT/GSK-3β signaling pathway in chronic intermittent hypoxia-induced insulin resistance.
Methods :
Forty SD rats were randomly divided into a control group(NC group) and an experimental group(CIH group), with 20 rats in each group. The NC group was placed in a normoxic environment for 12 weeks, while the CIH group was first subjected to intermittent hypoxia for 8 weeks, and then resumed normoxic rearing until the 12th week. Fasting blood glucose(FBG) and fasting insulin(FINS) were measured at baseline, week 8 and week 12, and liver tissues were taken for pathology and measurement of iNOS, p-IRS1ser 307, p-AKTser 473, GSK3β and GS levels, to compare the differences between groups.
Results:
t baseline, there was no significant difference in liver pathology between the two groups, and the observed indexes were not statistically significant(P>0.05); at 8 weeks, compared with the NC group, liver pathology in the CIH group showed significant disorganization of hepatic blood sinusoids and hepatocyte cords, obvious hepatocyte edema, smaller nuclei, increased lymphocyte infiltration, and a small number of fat vacuoles, significantly higher levels of FBG, FINS, insulin resistance index(HOMA-IR), iNOS mRNA, p-IRS1ser 307 protein, GSK-3β protein levels, and decreased p-AKTser 473 protein and GS protein levels, all of which were statistically significant(allP<0.05). IRS1ser 307 protein, GSK-3β protein levels were increased, p-AKTser 473 protein and GS protein levels were decreased, and the differences were statistically significant(allP<0.05); at 12 weeks, no lymphocyte infiltration was seen in the CIH group compared with that of the NC group and fat vacuoles significantly increased, and there was no improvement in the other pathological damage that had already occurred, and the levels of p-AKTser 473 protein significantly increased. AKTser 473 protein level significantly increased, p-IRS1ser 307 protein and GS protein levels were significantly reduced, all of which were statistically significant(allP<0.05), and the rest of the observational indexes were not statistically significant. Pearson′s correlation analysis showed that HOMA-IR of CIH group was significantly positively correlated with the levels of iNOS mRNA, p-IRS1ser 307 protein, and GSK-3β protein at 8 weeks(r=0.874, 0.817,0.872;allP<0.05), and significantly negatively correlated with the levels of p-AKTser 473 protein and GS protein(r=-0.886,-0.879;allP<0.05).
Conclusion
Chronic intermittent hypoxia can lead to hepatic pathological damage that cannot be reversed even by reoxygenation interventions and may mediate the development of insulin resistance by upregulating the IRS1/AKT/GSK-3β signaling pathway through the upregulation of iNOS mRNA expression.
2.Downregulation of Neuralized1 in the Hippocampal CA1 Through Reducing CPEB3 Ubiquitination Mediates Synaptic Plasticity Impairment and Cognitive Deficits in Neuropathic Pain.
Yan GAO ; Yiming QIAO ; Xueli WANG ; Manyi ZHU ; Lili YU ; Haozhuang YUAN ; Liren LI ; Nengwei HU ; Ji-Tian XU
Neuroscience Bulletin 2025;41(12):2233-2253
Neuropathic pain is frequently comorbidity with cognitive deficits. Neuralized1 (Neurl1)-mediated ubiquitination of CPEB3 in the hippocampus is critical in learning and memory. However, the role of Neurl1 in the cognitive impairment in neuropathic pain remains elusive. Herein, we found that lumbar 5 spinal nerve ligation (SNL) in male rat-induced neuropathic pain was followed by learning and memory deficits and LTP impairment in the hippocampus. The Neurl1 expression in the hippocampal CA1 was decreased after SNL. And this decrease paralleled the reduction of ubiquitinated-CPEB3 level and reduced production of GluA1 and GluA2. Overexpression of Neurl1 in the CA1 rescued cognitive deficits and LTP impairment, and reversed the reduction of ubiquitinated-CPEB3 level and the decrease of GluA1 and GluA2 production following SNL. Specific knockdown of Neurl1 or CPEB3 in bilateral hippocampal CA1 in naïve rats resulted in cognitive deficits and impairment of synaptic plasticity. The rescued cognitive function and synaptic plasticity by the treatment of overexpression of Neurl1 before SNL were counteracted by the knockdown of CPEB3 in the CA1. Collectively, the above results suggest that the downregulation of Neurl1 through reducing CPEB3 ubiquitination and, in turn, repressing GluA1 and GluA2 production and mediating synaptic plasticity impairment in hippocampal CA1 leads to the genesis of cognitive deficits in neuropathic pain.
Animals
;
Male
;
Neuralgia/metabolism*
;
Rats
;
Down-Regulation/physiology*
;
Ubiquitination/physiology*
;
Neuronal Plasticity/physiology*
;
Rats, Sprague-Dawley
;
CA1 Region, Hippocampal/metabolism*
;
Cognitive Dysfunction/metabolism*
;
RNA-Binding Proteins/metabolism*
;
Receptors, AMPA/metabolism*
3.Relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus
Hua XU ; Haijing YU ; Xueli GAO ; Qian WU ; Xiaoli WANG
Chinese Journal of Endocrine Surgery 2025;19(3):353-356
Objective:To study on the relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus.Methods:120 elderly patients with GDM were treated in Obstetrics Department of Yantai Yuhuangding Hospital from Jun. 2019 to Jun. 2021. The healthy women (58 cases) and normal pregnant women (58 cases) were selected as the control group at the same time, and the clinical data and serum samples of subjects were collected. The levels of serum SC, SV, FPG and HbAlc were measured, the HOMA-IR of the aged GDM was calculated, and the difference of serum index expression among the subjects in each group was analyzed. The relationship between serum SC, SV and insulin resistance was analyzed by Person correlation, and the risk factors of GDM were analyzed by Logistic regression.Results:: There was significant difference in HbAlc, HOMA-IR, FPG, SC, and SV among the three groups. The levels of each index in the elderly GDM group were significantly higher than those in the normal pregnancy group and the healthygroup ( P<0.05). Pearson analysis showed that SC ( P=0.558, 0.626, 0.672), SV ( P=0.576, 0.663, 0.696) was positively correlated with HbAlc, HOMA-IR, FPG in elderly patients with GDM ( P<0.001). Logistic regression analysis showed that SC, SV was risk factors for GDM in elderly pregnant women. Conclusion:There is a positive correlation between serum SC, SV level and insulin resistance, which are risk factors for GDM in elderly pregnant women.
4.Application of wearable devices in sarcopenia: a scoping review
Shuping LI ; Rui GAO ; Qiuchen YUAN ; Xiaolei WEI ; Xueli SHEN ; Tao LIANG
Chinese Journal of Modern Nursing 2025;31(19):2652-2657
Objective:To describe the application methods, types, and effects of wearable devices in sarcopenia.Methods:Following the guidelines for scoping reviews, a systematic search was conducted in PubMed, Cochrane Library, Web of Science, Embase, China National Knowledge Infrastructure, Wanfang Data, and VIP for literature related to wearable device applications in sarcopenia. The search period covered publications from database inception to August 1, 2024.Results:A total of 12 studies were included. Wearable devices in sarcopenia were primarily used for monitoring gait speed, surface electromyography, sleep duration, activity time, and supporting exercise interventions. Common types of wearable devices included activity trackers, smart belts, and smart wristbands. Their effectiveness, feasibility, and role in ensuring activity safety have been preliminarily demonstrated.Conclusions:The application of wearable devices in sarcopenia is still in its early stages but shows great potential. Currently, wearable devices are mainly used to monitor one or a few individual indicators. Future research should focus on developing multi-parameter monitoring devices and conducting large-scale, high-quality trials to validate their effectiveness in both monitoring and supporting exercise interventions. This will help promote their broader application and assist healthcare professionals in the early identification, diagnosis, prevention, and treatment of sarcopenia.
5.Relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus
Hua XU ; Haijing YU ; Xueli GAO ; Qian WU ; Xiaoli WANG
Chinese Journal of Endocrine Surgery 2025;19(3):353-356
Objective:To study on the relationship between serum levels of SC and SV and insulin resistance in elderly gestational diabetes mellitus.Methods:120 elderly patients with GDM were treated in Obstetrics Department of Yantai Yuhuangding Hospital from Jun. 2019 to Jun. 2021. The healthy women (58 cases) and normal pregnant women (58 cases) were selected as the control group at the same time, and the clinical data and serum samples of subjects were collected. The levels of serum SC, SV, FPG and HbAlc were measured, the HOMA-IR of the aged GDM was calculated, and the difference of serum index expression among the subjects in each group was analyzed. The relationship between serum SC, SV and insulin resistance was analyzed by Person correlation, and the risk factors of GDM were analyzed by Logistic regression.Results:: There was significant difference in HbAlc, HOMA-IR, FPG, SC, and SV among the three groups. The levels of each index in the elderly GDM group were significantly higher than those in the normal pregnancy group and the healthygroup ( P<0.05). Pearson analysis showed that SC ( P=0.558, 0.626, 0.672), SV ( P=0.576, 0.663, 0.696) was positively correlated with HbAlc, HOMA-IR, FPG in elderly patients with GDM ( P<0.001). Logistic regression analysis showed that SC, SV was risk factors for GDM in elderly pregnant women. Conclusion:There is a positive correlation between serum SC, SV level and insulin resistance, which are risk factors for GDM in elderly pregnant women.
6.Application of wearable devices in sarcopenia: a scoping review
Shuping LI ; Rui GAO ; Qiuchen YUAN ; Xiaolei WEI ; Xueli SHEN ; Tao LIANG
Chinese Journal of Modern Nursing 2025;31(19):2652-2657
Objective:To describe the application methods, types, and effects of wearable devices in sarcopenia.Methods:Following the guidelines for scoping reviews, a systematic search was conducted in PubMed, Cochrane Library, Web of Science, Embase, China National Knowledge Infrastructure, Wanfang Data, and VIP for literature related to wearable device applications in sarcopenia. The search period covered publications from database inception to August 1, 2024.Results:A total of 12 studies were included. Wearable devices in sarcopenia were primarily used for monitoring gait speed, surface electromyography, sleep duration, activity time, and supporting exercise interventions. Common types of wearable devices included activity trackers, smart belts, and smart wristbands. Their effectiveness, feasibility, and role in ensuring activity safety have been preliminarily demonstrated.Conclusions:The application of wearable devices in sarcopenia is still in its early stages but shows great potential. Currently, wearable devices are mainly used to monitor one or a few individual indicators. Future research should focus on developing multi-parameter monitoring devices and conducting large-scale, high-quality trials to validate their effectiveness in both monitoring and supporting exercise interventions. This will help promote their broader application and assist healthcare professionals in the early identification, diagnosis, prevention, and treatment of sarcopenia.
7.Characteristics of avian leukosis virus infection and polymorphism analysis of re-ceptor gene sequence in Qingyuan Ma chickens
Xiaoxue ZHENG ; Xueli SHU ; Hongmei WANG ; Mingchao GAO ; Mingcheng YU ; Yibin LI ; Ming LIAO ; Weisheng CAO
Chinese Journal of Veterinary Science 2024;44(7):1380-1386,1393
In order to understand the characteristics of avian leukosis virus(ALV)infection and the polymorphisms of receptor genes Tva,Tvb and NHE1 sequences in different Qingyuan Ma chick-en breeding farms,the isolation and identification of exogenous ALV virus,receptor gene amplifi-cation and sequencing analysis were carried out in five Qingyuan Ma chicken breeding farms.The results of virus isolation and subtype identification showed that the positive rate of exogenous ALV virus isolation in farms A,B and C was less than 5%,among which there was ALV-J infection alone in farm A,mixed infection with ALV-J and ALV-K in farm B,ALV-K infection a-lone in farm C,and no exogenous ALV infection was detected in farm D and E.Polymorphism anal-ysis of recipient genes showed that there were different frequencies of Tvar3,Tvar4 and Tvbr3 re-sistance alleles in A-E farms,and the distribution frequency of Tvar3 was 0.2-0.6,the distribution frequency of Tvar4 was 0.3-0.7,and the distribution frequency of Tvbr3 was 0.1-0.7.In addition,there were Tvar5 resistance alleles in both B and D farms,with a distribution frequency of 0.2.A to-tal of 18 SNP mutations occurred in the NHE1 receptor gene sequence,and further analysis showed that positions 1 279,1 361,1 369,1 406,1 442,and 1 912 were non-synonymous mutations,which could cause changes in amino acids.The study suggested that there were differences in the exogenous ALV and its subtypes among the five Qingyuan Ma chicken breeders,and each farm should have a more targeted and unique purification strategy.The distribution of Tva and Tvb re-sistance alleles at different frequencies,along with the occurrence of 6 non-synonymous mutations in the NHE1 gene,indicate that Qingyuan Ma chicken have the potential of genetic resistance breeding.
8.Antiosteoporosis effect of conventional treatment combined with Denosumab after percutaneous kyphoplasty for osteoporotic vertebral compression fractures
Chenyang WU ; Yiping GU ; Xueli QIU ; Huajian SHAN ; Xiang GAO ; Lide TAO ; Yingzi ZHANG ; Bingchen SHAN ; Xiaozhong ZHOU ; Jinyu BAI
Chinese Journal of Trauma 2024;40(9):787-792
Objective:To compare the antiosteoporosis effect of conventional treatment and conventional treatment combined with Denosumab after percutaneous kyphoplasty (PKP) for osteoporotic vertebral compression fractures (OVCF).Methods:A retrospective cohort study was conducted to analyze the clinical data of 211 patients with OVCF admitted to the Second Affiliated Hospital of Soochow University from September 2020 to September 2022. All the patients were female, aged 56-90 years [(71.4±8.1)years]. The bone mineral density T-score of the lumbar spine was (-2.6±1.0)SD before operation. Fracture segments included T 1-T 9 in 45 patients, T 10-L 2 in 146, and L 3-L 5 in 69. Of all, 174 patients were treated with single-segment surgery, 25 with two-segment surgery and 12 with surgery involving three or more segments. According to the wishes of the patients, 107 patients were treated with daily oral administration of calcium and active Vitamin D after PKP (conventional treatment group) and 104 patients with Denosumab combined with the conventional treatment after PKP (Denosumab therapy group). The bone mineral density T-scores of the lumbar spine of the two groups were compared before surgery and at the last follow-up. The visual analogue scale (VAS) and Oswestry disability index (ODI) before surgery, at 3 days, 6 months after surgery, and at the last follow-up were evaluated and the refracture rate after surgery was detected. Possible adverse effects after medication during anti-osteoporosis treatment were observed in two the groups. Results:All the patients were followed up for 12-24 months [(13.5±2.0)months]. Before surgery, the bone mineral density T-score of the lumbar spine was (-2.7±1.1)SD in the Denosumab therapy group and (-2.5±0.8)SD in the conventional treatment group ( P>0.05). At the last follow-up, the bone mineral density T-score of the lumbar spine was (-2.1±1.1)SD in the Denosumab therapy group, significantly higher than (-2.5±0.9)SD in the conventional treatment group ( P<0.05). In the Denosumab therapy group, the bone mineral density T-score of the lumbar spine at the last follow-up was significantly increased compared to that before surgery ( P<0.01), while there was no significant difference in the conventional treatment group ( P<0.05). Before surgery and at 3 days after surgery, the VAS scores and ODI values were (8.5±0.9)points, (2.8±0.8)points, 48.7±4.8 and 25.6±4.0 in the Denosumab therapy group, which was not statistically different from those in the conventional treatment group [(8.5±1.3)points and (2.8±0.9)points, 47.9±7.0 and 25.9±3.7] ( P>0.05). At 6 months after surgery and at the last follow-up, the VAS scores and ODI values were (2.2±0.8)points, (1.7±0.8)points, 24.2±3.6 and 23.2±4.1 in the Denosumab therapy group, significantly lower than those of the conventional treatment group [(2.8±0.9)points, (2.8±1.1)points, 26.4±3.2 and 27.3±4.0] ( P<0.01). The VAS scores at each time point after surgery in both groups decreased significantly compared with those before surgery ( P<0.05). The VAS scores continued to decrease after surgery in the Denosumab therapy group ( P<0.05), while no significant difference was found among those at different time points in the conventional treatment group ( P>0.05). The ODI values at each time point after surgery in both groups significantly decreased compared to those before surgery ( P<0.05). The ODI values continued to decrease after surgery in the Denosumab therapy group ( P<0.05), while in the conventional treatment group, no significant difference was found between those at 6 months after surgery and those at 3 days after surgery ( P>0.05) and they were improved at the last follow-up compared with those at 3 days after surgery ( P<0.05). The refracture rate after surgery was 6.7% (7/104) in the Denosumab therapy group, significantly lower than 16.8% (18/107) in the conventional treatment group ( P<0.05). No serious complications were observed during the antiosteoporosis period in either group. Conclusion:Compared with daily oral administration of Calcium and active Vitamin D after PKP, the conventional treatment combined with Denosumab after PKP can effectively increase the bone density, relieve pain continuously, improve functional restoration, and reduce the risk of refracture in OVCF patients.
9.Sleep status and influencing factors among higher vocational college students
Yong LI ; Xiaomei XIONG ; Fang HUANG ; Xia HUANG ; Xiu XU ; Zhaoxia HE ; Wenlong ZHANG ; Jiang ZHAO ; Xueli LIU ; Ru GAO
Sichuan Mental Health 2023;36(4):366-371
BackgroundSleep disorder can adversely affect human physical and mental health, with important implications for socioeconomic development. And higher vocational college students are subject to various pressures and stimuli exerted by academic studies, job seeking and family socioeconomic status, which may contribute to the onset insomnia symptom or sleep disorder. ObjectiveTo discuss the sleep status and influencing factors among higher vocational college students, and to provide references for improving their sleep status. MethodsIn January and February, 2022, a stratified random sampling method was utilized to enroll 3 300 students from five higher vocational colleges in Wenjiang district of Chengdu city. All the participants were assessed using Insomnia Severity Index (ISI), Patient Health Questionnaire Depression Scale-9 item (PHQ-9) and Generalized Anxiety Disorder Scale-7 (GAD-7). Pearson correlation analysis was conducted to test the correlation of ISI with PHQ-9 and GAD-7 scores, and the Logistic regression analysis was performed to identify the influencing factors of insomnia symptom in higher vocational college students. ResultInsomnia symptom was detected in 81.90% (2 497 / 3 300) of higher vocational college students. The detection rate of insomnia symptom among vocational college students yielded statistical difference in terms of gender, family socioeconomic status, being an only child or having siblings, degree of psychological pressure exerted by academic studies or job seeking, amount of hours spent on Internet per day, frequency of physical exercise, and the presence or absence of anxiety symptom and depression symptom (χ2=21.032, 22.172, 8.983, 75.939, 36.781, 32.350, 54.512, 86.561, P<0.01 or 0.05). Among higher vocational college students, ISI score was positively correlated with GAD-7 and PHQ-9 scores (r=0.620, 0.714, P<0.01), and GAD-7 score was positively correlated with PHQ-9 score (r=0.824, P<0.01). Gender, family socioeconomic status, degree of psychological pressure exerted by academic studies or job seeking, amount of hours spent on Internet per day, frequency of physical exercise, and the presence or absence of anxiety symptom and depression symptom were effective factors in predicting the insomnia symptom in higher vocational college students (P<0.01 or 0.05). ConclusionFemale gender, average and below average family socioeconomic status, moderate-to-severe degree of psychological pressure exerted by academic studies or job seeking, spending 2~5 h, 5~7 h or >7 h on Internet per day, inadequate physical exercise and presence of depressive symptom are the risk factors for insomnia symptom in higher vocational college students.[Funded by Sichuan Provincial Primary Health Development Research Center in 2022, North Sichuan Medical College (number, SWFZ22-C-89)]
10.Rapid evaluation of the early pathogen of severe Chlamydophila psittaci pneumonia by diagnostic bronchoscopy
Jingwen LI ; Shengquan LI ; Na LIU ; Tiantian SONG ; Lixiao ZHAO ; Xueli WANG ; Mengya GUO ; Yamei GAO ; Qiufeng WAN ; Sicheng XU
Chinese Critical Care Medicine 2023;35(1):37-42
Objective:To explore the rapid evaluation of the early pathogen of severe Chlamydophila psittaci pneumonia by bedside diagnostic bronchoscopy, so as to start effective anti-infection treatment before the results of macrogenome next generation sequencing (mNGS) test. Methods:The clinical data of three patients with severe Chlamydophila psittaci pneumonia who were successfully treated in the First Affiliated Hospital of Xinjiang Medical University, the First People's Hospital of Aksu District, and the First Division Hospital of Xinjiang Production and Construction Corps from October 2020 to June 2021 were retrospectively analyzed, including the rapid assessment of early pathogens by bedside diagnostic bronchoscopy and the use of antibiotics to start anti-infection treatment. These patients were successfully treated. Results:The three patients were male, aged 63, 45 and 58 years old, respectively. Before the onset of the penumonia, they had a clear medical history of bird exposure. The clinical manifestations mainly included fever, dry cough, shortness of breath and dyspnea. One case had abdominal pain and lethargy. The results of laboratory examination indicated that the peripheral blood white blood cell count (WBC) of two patients were high [(10.2-11.9)×10 9/L], the percentage of neutrophils increased (85.2%-94.6%) and the percentage of lymphocytes decreased (3.2%-7.7%) in all 3 patients after admission to hospital and entering into intensive care unit (ICU). The procalcitonin (PCT) of 3 patients increased after admission, and still increased when entering ICU (0.3-4.8 ng/L), so did C-reactive protein (CRP, 58.0-162.0 mg/L) and erythrocyte sedimentation rate (ESR, 36.0-90.0 mm/1 h). After admission, serum alanine transaminase (ALT) increased in 2 cases (136.7 U/L, 220.5 U/L), so did aspartate transaminase (AST) in 2 cases (249.6 U/L, 164.2 U/L). ALT (162.2-267.9 U/L) and AST (189.8-223.2 U/L) increased in 3 patients when they entered ICU. The level of serum creatinine (SCr) of 3 patients were normal after admission and entering ICU. The chest computed tomography (CT) findings of 3 patients were acute interstitial pneumonia, bronchopneumonia and lung consolidation, of which 2 cases were accompanied by a small amount of pleural effusion, and 1 case was accompanied by more regular small air sacs. Multiple lung lobes were involved, but mainly one lung lobe. The oxygenation index (PaO 2/FiO 2) of the 3 patients admitting to ICU were 100.0, 57.5 and 105.4 mmHg (1 mmHg ≈ 0.133 kPa), respectively, which met with the diagnostic criteria of moderate and severe acute respiratory distress syndrome (ARDS). All three patients received endotracheal intubation and mechanical ventilation. Under the bedside bronchoscope, the bronchial mucosa of 3 patients were obviously congested and edematous, without purulent secretion, and there was 1 case with mucosal hemorrhage. Three patients underwent bedside diagnostic bronchoscopy, and the evaluation result of the pathogen was that it might be atypical pathogen infection, so they were given moxifloxacin, cisromet and doxycycline intravenously, respectively, and combined with carbapenem antibiotics intravenously. After 3 days, the detection results of mNGS in bronchoalveolar lavage fluid (BALF) showed that only Chlamydia psittaci was infected. At this time, the condition was significantly improved, and PaO 2/FiO 2 was significantly increased. Therefore, the antibiotic treatment scheme remained unchanged, and mNGS only served to verify the initial diagnosis. Two patients were extubated on the 7th and 12th day of admission to the ICU, respectively, while one patient was extubated on the 16th day of admission to the ICU due to nosocomial infection. All 3 patients were transferred to the respiratory ward after the condition was stable. Conclusion:The bedside diagnostic bronchoscopy based on clinical characteristics is conducive to not only the rapid assessment of the early pathogens of severe Chlamydia psittaci pneumonia, but also effective anti-infection treatment before the returning of mNGS test results, which can make up for the lag and uncertainty of the mNGS test results.


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