1.Survival differences between endoscopic treatment and surgical treatment for patients with T1-2N0M0 duodenal neuroendocrine tumor
Bin BAI ; Xian SU ; Haibei XIN ; Minfeng ZHANG ; Hua XIAO ; Hui CAI
Chinese Journal of Clinical Medicine 2025;32(1):108-113
Objective To compare the long-term survival outcomes of patients with T1-2N0M0 duodenal neuroendocrine tumor (DNET) after endoscopic resection (ER) or surgical resection (SR). Methods Patients diagnosed with T1-2N0M0 DNET between January 1, 2004, and December 31, 2015, were extracted from the SEER database. Kaplan-Meier survival curve and log-rank test were used to compare overall survival (OS) rate and cancer-specific survival (CSS) rate between patients undergoing ER or SR. Propensity score matching (PSM) was used to reduce grouping differences, and multivariate Cox regression was used to analyze factors affecting OS and CSS before and after PSM. Results A total of 656 patients were included, with 457 in ER group and 199 in SR group. Before PSM, there was no significant difference in the 5-year OS rate between the ER and SR groups (88.9% vs 89.6%), but there was a significant difference in the 5-year CSS rate (99.3% vs 96.9%, P=0.017). Before PSM, multivariate Cox regression analysis showed advanced age was an independent risk factor for decreased OS (P<0.001). After PSM, there was no significant difference between the ER group (n=187) and SR group (n=187) in 5-year OS rate (90.2% vs 88.9%) or CSS rate (98.9% vs 96.7%). After PSM, multivariate Cox regression also showed advanced age was an independent risk factor for decreased OS, while resection method was not an independent factor for OS or CSS. Conclusions There is no significant difference in OS or CSS after endoscopic treatment and surgical treatments for patients with T1-2N0M0 DNET, and advanced age is an independent factor for OS.
2.Effects of intravenous and intraperitoneal routes on Babesia microti infections and splenic immune cells in BALB/c mice
Hanyin YANG ; Yuchun CAI ; Shuning YAN ; Yi XIN ; Ziran MO ; Bin XU ; Bin ZHENG
Chinese Journal of Schistosomiasis Control 2025;37(1):61-68
Objective To investigate the changes in the prevalence of Babesia microti infections, spleen morphology and proportions of splenic immune cells in BALB/c mice following intravenous and intraperitoneal injections, so as to provide insights into unraveling the immune regulatory mechanisms of Babesia infections. Methods Laboratory - maintained B. microti strains were prepared into whole blood samples with 10% prevalence of B. microti infection. A total of 75 BALB/c mice were randomly divided into three groups, including the normal control group, intravenous injection group, and intraperitoneal injection group, of 25 mice in each group. Mice in the intravenous and intraperitoneal injection groups were administered 100 μL of whole blood samples with 10% prevalence of B. microti infection, with the day of injection recorded as d0, and animals in the normal control group were given no treatments. Blood was sampled from mice in each group via the tail tip on d7, d14, d21, d28 and d35, and prepared into thin-film blood smears, and B. microti infection was observed in red blood cells. Five mice were randomly sampled from each group and sacrificed on d7, d14, d21, d28 and d35, and spleen was collected for measurement of spleen size and weight. In addition, splenic cells were isolated, and the proportions of CD3e+ T cells, CD45R+ B cells, CD49b+ nature killer (NK) cells, and F4/80+ macrophages were detected in CD45+ lymphocytes using flow cytometry. Results The prevalence of B. microti infection in the intravenous (22.80%) and intraperitoneal injection groups (44.82%) peaked on d7 (χ2 = 8.141, P < 0.01) and then rapidly decreased, and no parasites were observed on d35. The longest mouse spleen length [(32.91 ± 2.20) mm] and width [(9.82 ± 0.43) mm], and the greatest weight [(0.78 ± 0.10) g] were found on d14 in the intravenous injection group, and the longest spleen length [(32.42 ± 3.21) mm] and width [(10.25 ± 0.73) mm], and the greatest weight [(0.73 ± 0.09) g] were seen in the intra-peritoneal injection group on d21, d7 and d14, respectively. There were significant differences among the intravenous injection group, intraperitoneal injection group and the normal control group in terms of spleen length (F = 10.310, P < 0.05), width (F = 9.824, P < 0.05), and weight (F = 10.672, P < 0.05) on d21, and the mouse spleen length, width and weight were all significantly greater in the intraperitoneal injection group than in the intravenous injection group (allP values < 0.05). The proportions of splenic CD3e+ T cells [(60.60 ± 6.20)% and (39.68 ± 7.62)%], CD45R+ B cells [(43.32 ± 2.08)% and (49.53 ± 4.90)%], CD49b+ NK cells [(6.88 ± 1.34)% and (7.71 ± 1.59)%], and F4/80+ macrophages [(2.21 ± 0.29)% and (3.80 ± 0.35)%] peaked on d14, d21, d21 and d14 in the intravenous and intraperitoneal injection groups, respectively. There were significant differences in the proportions of CD3e+ T cells (F = 16.730, P < 0.05) and F4/80+ macrophages (F = 15.941, P < 0.05) among the intravenous injection group, intraperitoneal injection group and normal control group on d14, and a higher proportion of CD3e+ T cells and a lower proportion of F4/80+ macrophages were detected in the intravenous injection group than in the intraperitoneal injection group (both P values < 0.01). There were significant differences among the intravenous injection group, intraperitoneal injection group and normal control group on d21 in terms of proportions of splenic CD3e+ T cells (F = 9.252, P < 0.05), CD45R+ B cells (F = 14.349, P < 0.05), CD49b+ NK cells (F = 13.436,P < 0.05), and F4/80+ macrophages (F = 8.180, P < 0.05), and a higher proportion of CD3e+ T cells and lower proportions of CD45R+ B cells and F4/80+ macrophages were detected in the intravenous injection group than in the intraperitoneal injection group (all P values < 0.01). In addition, there was a significant difference in the proportion of CD3e+ T cells among the intravenous injection group, intraperitoneal injection group and normal control group on d28 (F = 9.772,P < 0.05), and a lower proportion of CD3e+ T cells was found in the intravenous injection group than in the intraperitoneal injection group (P < 0.01). Conclusions Both intraperitoneal and intravenous routes are effective to induce B. microti infections in BALB/c mice, and the prevalence of B. microti infections is higher in BALB/c mice through the intraperitoneal route than through the intravenous route. Intraperitoneal and intravenous injections with B. microti cause diverse spleen morphologies and proportions of splenic immune cells in mice, indicating routes of B. microti infections cause different impacts on immune response mechanisms in mice.
3.Dynamics of eosinophil infiltration and microglia activation in brain tissues of mice infected with Angiostrongylus cantonensis
Fanna WEI ; Renjie ZHANG ; Yahong HU ; Xiaoyu QIN ; Yunhai GUO ; Xiaojin MO ; Yan LU ; Jiahui SUN ; Yan ZHOU ; Jiatian GUO ; Peng SONG ; Yanhong CHU ; Bin XU ; Ting ZHANG ; Yuchun CAI ; Muxin CHEN
Chinese Journal of Schistosomiasis Control 2025;37(2):163-175
Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.
4.Effects of plateau environment on tear indexes and related anatomical structures of rabbits
Jinmei QIAN ; Bin HU ; Feie ZHANG ; Qi YANG ; Wenxue ZHU ; Jialing ZHANG ; Yan CAI
International Eye Science 2024;24(4):515-521
AIM: To investigate the effects of hypobaric hypoxia in plateau on tear indexes and related anatomical structures in rabbits.METHODS: A total of 18 healthy New Zealand rabbits were selected and randomly divided into plateau group and control group, with 9 rabbits(18 eyes)in each group. The plateau group was housed in the Simulated Climate Cabin for Special Environment of Northwest of China, simulating hypobaric hypoxia at an altitude of 6 000 m. The control group was housed in a clean animal room with atmospheric pressure and oxygen. Changes in the tear meniscus height and non-invasive tear break-up time were detected by using RHCT-1 corneal topographer dry eye comprehensive analysis system, changes in tear secretion was measured by Schirmer Ⅰ test, before intervention and on the 3, 7 d, 2 and 4 wk. Meanwhile, the changes in tear composition before and after intervention in the plateau environment were analyzed using Raman Spectroscopy. The histopathological changes of the lower lid conjunctiva, cornea, lacrimal gland, and Hardarian gland were observed by hematoxylin-eosin(HE)staining after 4 wk of intervention, and the expression of mucin 5AC(MUC5AC)in conjunctiva was detected by immunohistochemistry.RESULTS: Compared with the control group, Schirmer Ⅰ test, tear meniscus height, first and average non-invasive tear break-up time in the plateau group decreased significantly since 3 d, and the difference was significant with the extension of observation time(P<0.05). The above indexes increased from 2 wk. After 4 wk of intervention, the protein and lipid content of the tear composition of rabbits in the plateau group increased, and the nucleic acid content decreased compared with the pre-intervention period. Compared with the control group, rabbits in the plateau group showed thickening of corneal stromal edema, an increase in the number of conjunctival cup cells, increase in the level of expression of MUC5AC, an increase in the level of expression of MUC5AC, an atrophy and flattening of cytoplasm in lacrimal epithelial cells, an enlargement of glandular lumen, and no obvious destructive changes in the Hardarian glands.CONCLUSION: Acute plateau environment can destroy the homeostasis of rabbit ocular surface, so that the tear secretion and the tear film stability decreases, but within a certain period of time, rabbits undergo compensation with the habituation to the hypobaric hypoxia environment, which can increase the tear secretion to a certain extent and restore the tear film stability.
5.Risk factors and predictive model of complicating urosepsis after percutaneous nephrolithotripsy
Dawei ZHANG ; Gaoping CAI ; Yangyang TAN ; Yuxiang LI ; Cheng YANG ; Bin ZHANG ; Xukai YANG
Chongqing Medicine 2024;53(2):180-187
Objective To explore the risk factors of complicating urogenic sepsis after percutaneous nephrolithotripsy(PCNL)and construct a nomogram prediction model.Methods The data of 291 patients with stage 1 PCNL in 940 Hospital of Joint Logistics Support Force from October 2016 to October 2021 were retrospectively analyzed.The patients were divided into the sepsis group and non-sepsis group according to whether complicating urogenic sepsis after operation.The general data,stone-related data,operation-related data and laboratory detection related data were included.The independent risk factors were screened by the univariate and multivariate logistic regression analysis,and the nomogram prediction model was constructed.Results The results of univariate and multivariate logistic regression analysis showed that age≥60 years old(OR=6.438,95%CI:1.548-26.769),urinary leukocyte 3+(OR=5.651,95%CI:1.614-31.766),urinary nitrite positive(OR=7.117,95%CI:1.190-42.561),operation time≥90 min(OR=4.626,95%CI:1.137-18.817)and perfusion volume 30 L(OR=3.312,95%CI:1.090-10.061)were the independent risk factors of postoperative complicating urogenic sepsis.C-index of the constructed nomogram prediction model in the modeling samples was 0.937,the calibrated C-index was 0.914,and the model predictive efficien-cy was good.Conclusion Age ≥60 years old,urinary leukocyte 3+,urinary nitrite positive,operation time 90 min and perfusion volume ≥30 L are the independent risk factors for complicating urogenic sepsis after PCNL;the constructed nomogram prediction model has a good predictive efficiency for the occurrence of post-operative urogenic sepsis.
6.Inhibition of prohibitin 2 enhances the sensitivity of non-small cell lung cancer cell line A549 to erlotinib
Jing ZHANG ; Zigeng YANG ; Wenqin CAI ; Weiwei CAO ; Hongmei WEI ; Xixi XUE ; Bin WU
Basic & Clinical Medicine 2024;44(3):325-332
Objective To explore the effects of prohibitin 2(PHB2)on sensitivity of non-small cell lung cancer cell line A549 to erlotinib(Erl)and its potential mechanisms.Methods RACK1-specific small interfering RNA was transfected in A549 cells for knocking-down of PHB2.The effects of PHB2 inhibition on cell proliferation and apop-tosis induced by Erl were observed.The colocalization of microtuble-associated protein light chain 3 alpha(LC3)and mitochondria was visualized by MitoTracker staining and green fluorescent protein-microtuble-associated protein light chain 3 alpha(GFP-LC3)transfection.Cell proliferation was detected by 5-ethynyl-2′-deoxyuridine(EdU)staining.Cell colony formation was evaluated by colony forming assay.Apoptotic index of A549 cells was evaluated by TUNEL.Western blot was used to measure the expressions of PHB2 and LC3Ⅱ.Mitochondrial transmembrane potential,cytochrome c and respiratory chain complexⅠ/Ⅱ/Ⅴactivity were analyzed by the commercially availa-ble kits.Results Compared with the siPHB2 and siCtrl+Erl group,the EdU-positive A549 cells and the number of cell colonies decreased significantly(P<0.05),while the TUNEL-positive A549 cells increased significantly(P<0.05)in the siPHB2+Erl group.In addition,compared with the siPHB2 and siCtrl+Erl group,mitochondrial transmembrane potential and respiratory chain complexⅠ/Ⅱ/Ⅴactivity decreased significantly(all P<0.05)and the levels of cytochrome c increased in mitochondrial fractions(P<0.05)and decreased in cytosolic fractions(P<0.05)in the siPHB2+Erl group.Conclusions PHB2 inhibition significantly improves sensitivity of A549 cells to Erl,which may be explained by inhibition of PHB2-mediated mitochondrial autophagy.
7.Investigation and analysis of the incidence and influencing factors of thyroid dysfunction in patients with chronic kidney disease
Nan YANG ; Yahong CAI ; Bin LIU ; Yuan GUO ; Ruoyi LIAO
Chinese Journal of Endocrine Surgery 2024;18(1):74-78
Objective:To investigate the incidence of thyroid dysfunction in patients with chronic kidney disease and analyze the influencing factors.Methods:One hundred and ninety-eight patients with chronic kidney disease who were treated in Chronic Disease Management Department of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from Apr. 2021 to Apr. 2022 were selected, including 71 patients with abnormal thyroid function and 127 patients with normal thyroid function. The differences in TT3, FT3, TT4, FT4, and TSH between patients with abnormal thyroid function and those with normal thyroid function were analyzed. At the same time, the abnormal thyroid function of patients with different clinical characteristics and the influencing factors were analyzed. The intergroup differences were analyzed using t-test or χ2-test, and the influencing factors were analyzed using logistic regression analysis. Results:In one hundred and ninety-eight patients with chronic kidney disease, thyroid function abnormality occurred in 71 patients (35.86%), including two or more abnormal thyroid function indicators in 35 patients (49.30%). The total triiodothyronine (TT3), free triiodothyronine (FT3), total thyroxine (TT4) and free thyroxine (FT4) in patients with abnormal thyroid function were (1.02 ± 0.29) nmol/mL, (3.03 ± 0.88) pmol/L, (77.93 ± 20.02) nmol/mL and (11.02 ± 1.95) pmol/L respectively, which was significantly lower in patients with normal thyroid function (1.32±0.25) nmol/mL, (4.20±0.92) pmol/L, (93.30±19.28) nmol/mL and (13.54±1.82) pmol/ ( P<0.05), while thyroid stimulating hormone (TSH) was (3.14 ± 0.96) mIU/L, which was significantly higher than that in patients with normal thyroid function (1.84±0.89) mIU/L ( P<0.05). The incidence of thyroid dysfunction in female patients was 50.59% (43/85), It was significantly higher than 24.78% (28/113) of male patients (P <0.05) ; The incidence of thyroid dysfunction in patients aged 60 years was 49.55% (55/111), It was significantly higher than 18.39% (16/87) of the patients aged <60 years ( P<0.05) ; The incidence of thyroid dysfunction in patients with 1-year duration of disease was 71.43% (30/42), It was significantly higher than 25.28% (41/156) of patients with a course of disease <1 year ( P<0.05) ; The incidence of thyroid dysfunction in patients with clinical stage G 4 to 5 was 50.62% (41/81), It was significantly higher than 25.64% (30/117) of patients in G1~3 stages ( P<0.05) ; The incidence of thyroid dysfunction in patients with diabetes was 74.36% (29/39), This was significantly higher than 26.42% (42/159) in patients without diabetes mellitus ( P<0.05). Logistic regression analysis showed that gender, age, course of disease and clinical stage were the influencing factors of thyroid dysfunction in patients with chronic kidney disease ( P<0.05) . Conclusion:A high proportion of patients with chronic kidney disease have abnormal thyroid function, which is affected by the patient's sex, age, course of disease and clinical stage,clinical attention should be paid to targeted intervention to prevent the incidence of thyroid dysfunction in chronic kidney disease population.
8.Effect of perioperative precision nursing intervention on elderly patients with femoral neck fractures
Yingying HE ; Jinfen CHEN ; Xiaoli ZHANG ; Zuolei JING ; Guihuan LI ; Qiongzhu CAI ; Bin ZHANG
Modern Hospital 2024;24(2):325-328
Objective To investigate the impact and significance of a precision nursing plan during the surgical manage-ment of femoral neck fractures in elderly patients.Methods From May 2022 to May 2023,70 patients with femoral neck frac-tures,including medium-sized neck and head-type,were randomly divided into a control group and an observation group equally.The two groups were managed with routine nursing interventions and specific nursing interventions,respectively.The two groups were compared in terms of the psychological status,complications,hip joint functions,and prognostic effects.Results After the intervention,the psychological status scores of both two groups significantly decreased.The observation group showed lower psy-chological status scores,indicating better psychological status compared to the control group(P<0.05).The rate of postopera-tive complications in the observation group was significantly lower than that of the control group(P<0.05).Additionally,the scores of hip joint function in the observation group were higher than those of the control group(P<0.05).At the time of dis-charge,the scores of self-care ability in both two groups had increased,and the scores of femoral head necrosis were significantly decreased after 3 months of interventions(P<0.05).Furthermore,the self-care ability scores in the observation group were higher,and the scores of femoral head necrosis were lower compared to those of the control group,indicating that the prognosis of the observation group was better than that of the control group(P<0.05).Conclusion The precise nursing interventions could effectively enhance the prognosis of elderly patients with femoral neck fractures and improve their self-care ability.
9.Characteristics of schizophrenic patients using long-acting antipsychotic medications
GE Xiaolei ; ZHANG Weibo ; CHEN Chunmei ; ZHU Youwei ; LIU Yanli ; XIE Bin ; CAI Jun ; ZHU Yi
Journal of Preventive Medicine 2024;36(5):412-415
Objective:
To investigate the characteristics of schizophrenic patients using long-acting antipsychotic medications, so as to provide the basis for applicable population of long-acting antipsychotic medications.
Methods:
Data of schizophrenic patients using long-acting antipsychotic medications in Shanghai City from June 2020 to June 2022 were collected through Shanghai Mental Health Information Management System, and demographic characteristics, illness and medication use of patients were descriptively analyzed.
Results:
A total of 2 684 schizophrenic patients using long-acting antipsychotic medications were included in the study, had a mean age of (46.92±12.39) years, with 1 246 males (46.42%) and 1 438 females (53.58%). There were 1 397 unemployed cases, accounting for 52.05%; 1 429 cases with an educational level in junior high school or below, accounting for 53.24%; 1 301 unmarried cases, accounting for 48.47%; 832 cases in poverty, accounting for 31.00%. The caregivers of patients were mainly their parents, with 1 507 cases accounting for 56.15%. The courses of illness were mainly ≤10 years and >10-20 years, with 860 cases each, both accounting for 32.04%; 1 963 cases with incomplete self-awareness, accounting for 73.14%; 1 570 cases hospitalized at least once, accounting for 58.49%. There were 2 486 cases with continuous medication, accounting for 92.62%. The main method of taking medication was given by others, with 1 947 cases accounting for 72.54%. The medication adherence was mainly taking medication on time and in the right amount, with 2 437 cases accounting for 90.80%.
Conclusion
The main characteristics of schizophrenic patients using long-acting antipsychotic medications are young, unmarried, and unemployed adults, with incomplete self-awareness, continuous medication and medication given by others.
10.Study of The Y-STR Multiplex Microfluidic Chip Rapid Amplification System
Dao-Yu WANG ; Qun WAN ; Bin ZHUANG ; Li-Jian ZHAO ; Jun-Ping HAN ; Cai-Xia LI
Progress in Biochemistry and Biophysics 2024;51(3):696-705
ObjectiveAt present, the matching reagents of commercially available rapid DNA instruments based on microfluidics chip technology are autosome short tandem repeat (STR) individual identification reagents. The non-recombining part of the human Y chromosome is widely used in forensic DNA analysis, particularly in cases where standard autosomal DNA profile is uninformative. Y-STR loci are useful markers to identify males and male lineages in forensic practice. In order to achieve rapid and fully integrated detection ofY-STR loci, this study constructed the RTyper Y27 microfluidic chip rapid detection system and validated the performance of this system. MethodsThe system was verified and evaluated by sensitivity, success rate, typing accuracy, peak height balance, sizing precision and accuracy, mock case sample tests, mixture detection ability, and inhibition tolerance. ResultsComplete Y-STR profiles can be obtained when the template amount of DNA standard 9948 was ≥8 ng, the number of blood cards was ≥3 pieces, and the number of oral swab scrapings was≥7 times. The success rate of fully integrated detection was 91.52%, and the concordance rates was 99.74% for 165 testing samples. The success rate of 115 blood spots in these samples was 90.43%, with a typing accuracy of 99.65%, the success rate of 50 buccal swabs was 94%, with a typing accuracy of 99.92%. There was no significant difference in typing accuracy between blood spots and buccal swab samples. The peak height ratio between different fluorescence channels was 89.81%. The standard deviation of allelic ladder for 10 runs was within 0.5 bp. The size differences between allele and corresponding allele in allelic ladder was within 0.5 bp. The maximum precision CV values within and between batches were 0.48% and 0.68%, respectively, which were lower than 15%. These data indicate that the system has good accuracy and precision. The system was capable of accurately typing oral swabs, blood cards, saliva cards, cigarette butts, blood swabs and seminal stains. Complete Y-STR profiles can be obtained and distinguish at the 1∶3 ratio of minor and major contributors in artificial male DNA mixtures. Complete Y-STR genotyping can be obtained under the interference of inhibitors, such as different concentrations of humic acid (50-400 mg/L), indigotin (20-100 nmol/L) and hemoglobin (100-500 μmol/L). ConclusionIn this study, the RTyper Y27 microfluidic chip rapid amplification system is combined with the Quick TargSeq 1.0 integrated system, and the Y-STR profile can be obtained in approximately 2 h. Through a series of verification experiments, the results show that the system has good repeatability, accuracy and stability, can meet the on-site Y-STR detection requirements, and can be used in forensic practice.


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