1.Isolation,culture and differentiation of human urine-derived stem cells into smooth muscle cells
Jiahui CHEN ; Xiaoqi DAI ; Yangang XU ; Yuanchao LI ; Mei HUANG ; Yifei ZHAN ; Yuxuan DU ; Liuqiang LI ; Yaochuan GUO ; Jun BIAN ; Dehui LAI
Chinese Journal of Tissue Engineering Research 2025;29(19):4076-4082
BACKGROUND:Traditional methods of urinary tract reconstruction are limited by donor scarcity,high complication rates,and suboptimal functional recovery.Tissue engineering strategies offer new directions in this field.Since the urinary tract is mainly composed of muscle tissue,the key is to find suitable seed cells and efficiently induce them to differentiate into smooth muscle cells.Comparative studies on the efficacy of different smooth muscle cell induction regimens are still lacking. OBJECTIVE:To isolate,culture,and identify human urine-derived stem cells,and to compare the effects of two different induction protocols. METHODS:Human urine-derived stem cells were isolated from urine samples of 11 healthy adult volunteers by multiple centrifugations.Surface markers were identified by flow cytometry.The multi-directional differentiation potential of human urine-derived stem cells was verified through osteogenic and adipogenic differentiation.Differentiation was induced by transforming growth factor-β1 or transforming growth factor-β1 combined with platelet derived growth factor for 14 days.Immunofluorescence staining and western blot assay were employed to compare the expression differences of smooth muscle-specific proteins(α-SMA and SM22). RESULTS AND CONCLUSION:(1)Urine-derived stem cells were successfully isolated from the eight urine samples of healthy people.These cells exhibit a"rice grain"-like morphology and possess a robust proliferative capacity.(2)Urine-derived stem cells exhibited high expression of mesenchymal stem cell surface markers(CD73,CD90,and CD44)and extremely low expression of hematopoietic stem cell surface markers(CD34 and CD45).These cells did not express CD19,CD105,and HLA-DR.(3)After osteogenic and adipogenic differentiation,the formation of calcium nodules and lipid droplets was observed,with positive staining results from Alizarin Red S and Oil Red O staining.(4)After 14 days of smooth muscle induction culture,immunofluorescence staining revealed that the smooth muscle differentiation rate of urine-derived stem cells treated with a combination of transforming growth factor-β1 and platelet derived growth factor was significantly higher compared to those treated with transforming growth factor-β1 alone(P<0.005).(5)After 14 days of smooth muscle induction culture,western blot assay further demonstrated that the expression levels of α-SMA and SM22 in the transforming growth factor-β1/platelet derived growth factor group were significantly elevated compared to those in the transforming growth factor-β1 only group(P<0.005).These findings confirm that urine-derived stem cells can be non-invasively isolated using multiple rounds of centrifugation.Compared with transforming growth factor-β1 alone,the combination of transforming growth factor-β1 and platelet derived growth factor can improve the efficiency of inducing urine-derived stem cells to differentiate into smooth muscle cells.
2.Effect of activating α7 nicotinic acetylcholine receptor on learning and memory function and microglia in traumatic brain injury model rat
Juan DU ; Han ZHANG ; Yuxuan YANG ; Bo ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(2):116-121
Objective:To explore the effects of activating α7 nicotinic acetylcholine receptor(α7nAChR)on cognitive function and polarization of hippocampal microglia in traumatic brain injury (TBI) rats.Methods:Totally 36 male SD rats with 6-8 weeks old were randomly divided into Sham group ( n=12), TBI group ( n=12), TBI+ α7nAChR agonist group ( n=6) and TBI+ α7nAChR antagonist group( n=6). The TBI model was established by the " free fall impact" method. From the 4th to 6th day after modeling, mice in the TBI+ α7nAChR agonist group were intraperitoneally injected with α7nAChR agonist PNU-282987 (3 mg/kg). Rats in TBI+ α7nAChR antagonist group were intraperitoneally injected with α7nAChR antagonist methyllycaconitine citrate (5 mg/kg) first, then 45 minutes later they were injected with α7nAChR agonist PNU-282987 (3 mg/kg). Rats in the TBI group and Sham group were intraperitoneally injected with an equal volume of 0.9% sodium chloride solution. Morris water maze test was used to evaluate the learning and memory function of rats. Immunofluorescence staining was used to observe the ionized calcium binding adapter molecule 1 (Iba-1)(a marker for microglia) and arginase 1 (Arg-1)(a marker for M2 microglia). Western blot was used to detect the protein level of Arg-1 in hippocampal tissue. Statistical analysis was performed using GraphPad Prism 9 software. Independent sample t test was used for comparison between two groups, one-way ANOVA was used for comparison among multiple groups, and Tukey test was used for multiple comparison. Results:The results of the water maze test showed that after 7 days of modeling, there was a statistical difference in the escape latency among the 4 groups of rats ( F=6.134, P<0.05). There was no statistical difference in the escape latency between the TBI group and the TBI+ α7nAChR antagonist group( P>0.05), but the both were higher than that of the Sham group (both P<0.05). The escape latency of the TBI+ α7nAChR agonist group((31.87±9.01)s) was shorter than that of the TBI group((56.75±2.62)s) and the TBI+ α7nAChR antagonist group((60.00±0.00)s) (both P<0.05). The results of immunofluorescence staining showed that there were statistical differences in the fluorescence intensity and cell numbers of Arg-1 + /Iba-1 + among the four groups ( F=17.37, 9.33, both P<0.05). The immune fluorescence intensity (0.27±0.03) and cell numbers (21.67±4.41) of Arg-1 + /Iba-1 + in the TBI+ α7nAChR agonist group were higher than those in the TBI group((0.14±0.03), (11.33±2.60)) and TBI+ α7nAChR antagonist group((0.10±0.03), (7.67±1.20)) (all P<0.05). The results of Western blot showed that there was a statistical difference in the level of Arg-1 protein in hippocampus among the 4 groups ( F=8.323, P=0.001). There was no significant difference in the level of Arg-1 protein between the TBI group and the TBI+ α7nAChR antagonist group( P>0.05), and the level of Arg-1 protein in the TBI+ α7nAChR agonist group(1.06±0.22) was higher than that in the TBI group(0.60±0.13) and TBI+ α7nAChR antagonist group(0.35±0.10) (both P<0.05). Conclusion:Activating α7nAChR can promote the polarization of M2 type microglia in rat hippocampal tissue and improve the learning and memory function of TBI rats.
3.A study on the animal model preparation and early histological changes of trans-sutural distraction osteo-genesis
Zhihe ZHAO ; Yufan ZHANG ; Wenhui ZHANG ; Yicheng CHEN ; Xuelian JIA ; Shanluo ZHOU ; Fuding SHEN ; Yuxuan DU ; Yunpeng LI ; Liang KONG
Journal of Practical Stomatology 2024;40(2):173-179
Objective:To establish an animal model of trans-sutural distraction osteogenesis in SD rats.Methods:A self-designed V-shaped distraction device(distractor)was fabricated with the traction force(N)of 0,1.3,2.2,3.0,4.3 and 5.0 corresponding to the distraction length(mm)of 5,4,3,2,1 and 0 respectively,meeting the trans-sutural distraction osteogenesis requirements in skull of 5-week-old SD rats.The distractor was plased into the sagittal suture of 12 SD rats.Continuous sampling was conducted 1,3,5 and 7 days respectively(n=3)after operation.The tissue changes in the trans-sutural distraction area were observed by HE and Masson's trichrome staining.Inflammation levels were determined using Arg-1 immunofluorescence staining.The early angiogenesis was clarified through co-staining with CD31 and EMCN.Results:A stable trans-sutural distraction osteogenesis model was estab-lished,5 mm distraction osteogenesis width was observed completely within 7 days of distraction.Significant new bone formation was observed at 7 days after operation.Arg-1 expression increased and was concentrated at the bone margins,overlapping with the areas of new bone formation.EMCN expression gradually decreased,and by day 7 CD31 was predominant,indicating the basic maturation of blood vessels.Conclusion:This study successfully constructed a stable and effective trans-sutural distraction osteogenesis animal model,and provides an experimental basis for the investigation of its early continuous histological changes.
4.Impact of emotions on cancer risk:a Mendelian randomization study
Qin DU ; Yuxuan ZHU ; Yize GUO ; Nanxin XU ; Di LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(3):376-382
Objective To investigate the effects of emotions(subjective well-being,depressed effect,worry,and guilt)on cancer(colorectal cancer,hepatic cancer,thyroid cancer,lung cancer,and breast cancer).Methods Two-sample bi-directional Mendelian randomization(MR)method was adopted.All data were based on summary data from genome-wide association studies(GWAS).Inverse variance weighting(IVW)was used to generate the main results,and weighted median(WM)and MR-Egger methods were employed to calculate supplementary results.The outcome measure was odds ratio(OR),and sensitivity analysis was conducted.Results For depressed effect,a significant association with lung cancer(OR=1.005,95%CI:1.001-1.009,P=0.015)was found.For worry,a significant association with breast cancer(OR=1.199,95%CI:1.011-1.423,P=0.038)was observed.For guilt,a significant association with thyroid cancer(OR=2.083,95%CI:1.080-4.017,P=0.029)was identified.After removing all potentially pleiotropic SNPs detected by MR PRESSO,the association between worry and breast cancer showed no statistical difference(P=0.064),while the association between worry and colorectal cancer remained significant(OR=0.739,95%CI:0.571-0.956,P=0.021).No causal relationship was found between cancer and emotions.Conclusion There is a causal relationship between depression and increased lung cancer incidence,guilt and increased thyroid cancer incidence,as well as anxiety and decreased colorectal cancer incidence.
5.Correlation of the distribution and morphology of breast calcification with the risk of breast malignant tumors
Yuxuan ZHU ; Qin DU ; Yize GUO ; Nanxin XU ; Di LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(6):982-987
[Objective] To analyze the correlation of the distribution and morphological characteristics of breast calcification with the risk of breast malignant tumors, so as to further reveal the important value of the calcification in diagnosis of breast malignant tumors. [Methods] A total of 108 patients who had received concurrent puncture biopsy and surgical treatment of breast calcification at The Second Affiliated Hospital of Xi’an Jiaotong University from January 1, 2019 to December 31, 2019 were recruited, and their imaging data and postoperative pathological results were analyzed. We divided the distribution of breast X calcification into two categories based on segmental/linear calcification and regional/diffuse calcification. We classified the morphology of breast X calcification into punctate, small branching, polymorphic, amorphous, and coarse ones. We then explored the distribution, morphology, and BI-RADS classification of tumor calcification under breast X-ray, and its correlation with the occurrence, age, menopausal status, hormone receptors, and molecular typing of breast malignant tumors. [Results] Among the 108 patients enrolled in the study, 21 cases were malignant and 87 cases were benign. The results showed that the distribution of calcification was segmental/linear (χ2=11.2, P=0.003) and the calcification morphology was of small branching calcification (χ2=9.3, P=0.046). The detection rate in malignant tumors was significantly higher than that in benign tumors. The distribution and morphology of calcification were not significantly correlated with ER, PR, HER2, Ki67, pathological type, or molecular typing (P>0.05). Logistic regression analysis showed that , calcification distribution was segmental/linear [OR=19.94(3.061-129.9), P=0.002], calcification morphology was small branching [OR=3.906(1.141-13.37), P=0.030], and B-RADS classification was 4 grade and above [OR=39.99(2.703-591.7), P=0.007], which were closely related to the occurrence of malignant tumors. [Conclusion] For patients with breast tumors in which calcification can be detected under mammography, their imaging characteristics are closely related to the occurrence of malignant tumors. The distribution of calcification is segmental/linear, and the morphology of calcification is small branching/amorphous, which is more likely to be a malignant tumor and closely related to the occurrence of breast cancer.
6.Epidemiological distribution of mosaic loss of chromosome Y in adult men in 10 areas in China and its prospective association with lung cancer
Yuxuan ZHAO ; Mingyu SONG ; Jun LYU ; Canqing YU ; Pei PEI ; Huaidong DU ; Junshi CHEN ; Zhengming CHEN ; Liming LI ; Dianjianyi SUN
Chinese Journal of Epidemiology 2024;45(1):56-62
Objective:To detect the prevalence of mosaic loss of chromosome Y in adult men in ten study areas in China, describe the epidemiological distribution of mosaic loss of chromosome Y (mLOY) carriers and assess its prospective association with lung cancer.Methods:Based on the data from baseline survey, genetic analysis and follow-up (as of December 31, 2018) from China Kadoorie Biobank, we used Mosaic Chromosomal Alterations pipeline to detect mLOY carriers in 10 areas in China and described the epidemiological characteristics of mLOY carriers in adult men, including age, area distribution, lifestyle and disease history. We used multivariate logistic regression model to identify the potential relevant factor of mLOY. Cox proportional hazard regression model was fitted to assess the prospective association of mLOY with lung cancer. Stratification analysis were conducted to evaluate the potential modification effects of smoking and age. We also conducted mediation analysis to assess the mediating effect of mLOY in the association between smoking and lung cancer.Results:A total of 42 859 adult men were included in our analysis, in whom 2 458 mLOY carriers were detected (5.7%). The detection rate increased with age ( P<0.05). The detection rate was higher in urban area (7.3%±0.2%) than that in rural area (4.7%±0.1%). The results of logistic regression analysis indicated that smoking might be a risk factor for the detection of mLOY ( OR=1.49, 95% CI:1.36-1.64). After follow-up for average 11.1 years, 1 041 lung cancer cases were observed. The prospective analysis showed that mLOY carriers had an increased risk for lung cancer by 24% compared with non-mLOY carriers ( HR=1.24, 95% CI:1.01-1.52) and expanded mLOY carriers (mLOY cell proportion ≥10%) had an increased risk for lung cancer by 50% ( HR=1.50, 95% CI:1.13-2.00). Stratification analysis showed no modification effects of smoking and age in the association between mLOY and lung cancer (interaction P>0.05). Mediation analysis showed that mLOY could be a mediating factor in the association between smoking and lung cancer, the estimated effect was 0.09 (0.01-0.17). Conclusions:There were significant differences in the detection rate of mLOY in adult men with different social-economic characteristics and lifestyles in ten areas in China. Besides, mLOY carriers, especially expanded mLOY carriers, had increased risk for lung cancer and mLOY might be a mediating factor in the association between smoking and lung cancer.
7.Qualitative study on residents' cognition of commercial long term insurance in Tianjin
Yue DU ; Yuxuan BAI ; Jinyan CHEN ; Li SUN ; Jiawei NI ; Yaxuan XU ; Zixiu XU ; Ruichen SUN ; Na TANG
Chinese Journal of Modern Nursing 2024;30(13):1795-1799
Objective:To understand the residents' cognition of commercial long-term insurance in Tianjin so as to provide a reference for improving service effect and quality of long-term insurance.Methods:A total of 31 residents from six districts of Tianjin were selected for semi-structured interviews from December 2022 to February 2023 by the convenient sampling method, and the data were analyzed by Colaizzi 7-step analysis method.Results:Three themes were extracted, namely, inadequate interpretation and promotion of commercial long-term insurance, immature development of commercial long-term insurance, and the ability of commercial long-term insurance to help caregivers reduce their family burden.Conclusions:The residents of Tianjin have a low awareness of commercial long-term insurance, which cannot meet the needs of most residents. It is urgent to strengthen their publicity further and improve their system.
8.Fibrinolytic effect of curdione in dissolving deep vein thrombosis in rats
Hanbing SHI ; Teng ZHANG ; Shan DU ; Yuxuan GUAN ; Yan HUANG ; Quan XIA
China Pharmacist 2024;28(11):424-430
Objective To investigate whether curdione inhibits the formation of deep vein thrombosis(DVT)in rats through thrombolytic effects.Methods Twenty-five rats were randomly divided into five groups,the normal control group,the sham operation group,the model group,the low dose group of curdione(60 mg/kg)and the high dose group of curdione(120 mg/kg),with five rats in each group.Using the inferior vena cava ligation model,blood was taken from the pulmonary aorta in a sodium citrate anticoagulant tube one week after drug administration.The coagulation and fibrinolytic indexes Fibrin Degradation Products,(FDP),D-dimer(D-D),plasminogen activator inhibitor(PAI)-1 and plasminogen(PLG)were detected in each group;the thrombus weight/weight-length ratio was detected;and the formation of venous thrombus in the lower limbs of rats was detected by hematoxylin-eosin(HE)staining nuclear factor;NF-κB protein expression in inferior vena cava tissues was dectected by Western Blot assay.Results There was no significant difference in plasma FDP levels among groups of rats(P>0.05);compared with the model group,the plasma FDP and D-D levels of rats in each dosing group were increased,the plasma PAI-1 and PLG levels were elevated,and the fibrinolytic effect was better in the group with high dose of curdione administration than in the group with low dose of curdione administration(P<0.05);compared with the model group,the thrombus weight/weight-length ratio,the thrombus mass and NF-κB protein expression in the curdione dosing group were decreased.Conclusion Curdione can inhibit the formation of DVT in rats by influencing the fibrinolytic regulators.
9.Analysis on the current status in fundamental construction of clinical laboratories in China
Yuxuan DU ; Wei WANG ; Zhixin ZHANG ; Na DONG ; Zhiguo WANG
Chinese Journal of Laboratory Medicine 2024;47(9):1052-1058
Objective:To investigate the current status on fundamental construction of clinical laboratories in China.Methods:A web-based external quality assessment (EQA) system was used to collect basic information from the clinical laboratories that participated in EQA program of National Center for Clinical Laboratory in 2023. The indicators include the construction of laboratory information systems (LIS), laboratory building area, human resource, and the number of clinical testing item. The comparisons of the indicators between different types of laboratories were tested.Results:A total of 14 405 clinical laboratories submitted the required information in 2023. Guangdong province accounted for the highest proportion of the numver of laboratories, which is 10.6% (1 533/14 405). The building area and staff numbers in the independent clinical laboratory were higher than other institutions, which were 1 400 (850, 2 196) m 2 and 20 (12, 35), respectively. The building areas, the staff numbers and the number of clinical testing item of tertiary hospitals and general hospitals were significantly higher than other hospitals. The building areas of the tertiary hospitals and the general hospitals were respectively 1 050 (600, 1 800) m 2 and 1 000 (500, 1 600) m 2, and the staff numbers were respectively 30 (19, 45) and 26 (17, 39). Besides, the number of clinical testing item were 287 (189, 388) and 285 (198, 378), respectively. The proportion of staff in hospitals with master′s degrees or higher titles was 18.2% (52 138/286 855). Meanwhile, 17.4% (49 280/282 528) of the staff had higher academic title. It showed that 24.5% (3 528/14 405) of the clinical laboratories still have no LIS. Conclusion:The fundamental construction in different type of clinical laboratories varied significantly in China. The clinical laboratories should strengthen the construction of information systems and pay more attention to human resources.
10.Analysis of external quality assessment results of quality indicators for national emergency examination turnaround time
Zhixin ZHANG ; Wei WANG ; Yuxuan DU ; Jiali LIU ; Bingquan CHEN ; Zhiguo WANG
Chinese Journal of Laboratory Medicine 2023;46(7):719-724
Objective:To establish preliminary quality specifications for emergency examination turnaround time (TAT).Methods:The National Center for Clinical Laboratories organized 31 provinces (autonomous regions and municipalities directly) and Xinjiang production and Construction Corps centers to launch a synchronous Quality Indicators (QIs)-External Quality Assessment (EQA) program and the collected data were reported via developed online EQA system. The essential information of the clinical laboratories, the data of pre-examination and intra-laboratory TAT quality indicators of emergency departments at each specialty (biochemistry, automatic immunity, three routines tests and coagulation) and four specific tests (blood potassium, troponin I/T, white blood cell count and international normalized ratio (INR)) were collected from 2019 to 2021. TAT returned the median and 90th percentile ( P90) of the specified month were calculated. The median (lower quartile, upper quartile) of the TAT returned laboratories were calculated and second result grading statistics for 2021 (2 422 tertiary hospital and 5 088 secondary hospital) were performed to understand the difference of pre-examination and the laboratory TAT between different tertiary hospitals. Results:From 2019 to 2021, there were 9 540 laboratories, 9 709 laboratories and 10 653 returned laboratories. The pre-examination TAT of each specialty was similar, and the results were relatively stable. The median distribution was about 15 (10, 30) min, and the monthly P90 distribution was about 20 (10, 30) min. The distribution results of the median intra-laboratory TAT in each specialty were as follows: automatic immunity≥biochemistry>coagulation>three routine tests. The distribution of the latest (second result in 2021) survey results of each specialty were as follows: automatic immunity 53 (30, 60) min, biochemistry 45 (30, 60) min, coagulation 30 (23, 40) min, and three routine tests 20 (11, 30) min. The median results of monthly P90 of intra-laboratory TAT were as follows: 60 min for automatic immunity and biochemistry specialty, about 38 min for coagulation specialty, and about 27 min for three routines tests. The hierarchical statistical results showed that the monthly P90 distribution of laboratory TAT of the pre-examination and intra-laboratory TAT from the tertiary hospital was higher than that of the secondary hospital. The pre-examination TAT of each specialty of the tertiary hospital/secondary hospital was as follows: biochemistry 35 (22, 60)/20 (11, 30) min, automatic immunity 33 (20, 60)/20 (10, 30) min, three routine tests 30 (20, 49)/20 (10, 30) min and coagulation 31 (20, 58)/20 (10, 30) min, the intra-laboratory TAT of each specialty of the tertiary hospital/secondary hospital was as follows: biochemistry 65 (50, 91)/60 (40, 70) min, automatic immunity 75 (55, 113)/60 (40, 90) min, three routine tests 30 (23, 38)/28 (19, 30) min and coagulation 53 (36, 72)/35 (30, 57) min. In terms of the distribution results of the median of intra-laboratory TAT of the four specific tests, 96.76% (9 484/9 801) of the blood potassium and 95.96% (8 733/9 101) of the troponin I/T medical institutions were TAT within 69 min in the laboratories, 95.34% (9 679/10 152) of the white blood cell count medical institutions were TAT within 31 min in the laboratories, and 98.85% (9 462/9 572) of the INR medical institutions were TAT within 66 min in the laboratories. Conclusions:This survey provides a preliminary quality specification for the emergency department turnaround time at each specialty. Lower quartile, median and upper quartile of the monthly P90 at the tertiary and secondary hospitals can be used to define the best, appropriate and minimum performance levels, respectively.

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