1.Isolation and differentiation of stem cells derived from human placenta into cardiomyocytes
Yanqiu YU ; Kun LI ; Yunan JIN ; Wei YUN ; Lili DU ; Xiaomei LU ; Haiqin REN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To isolate the stem cells from human placenta and induce them into cardiomyocytes. METHODS: Stem cells were isolated from human placenta and characterized by morphologic analysis. The “hanging drop” methods were used to inducte stem cells into cardiomyocytes. The expressions of atrial natriuretic factor(Anf) and ?-myosin heavy chain(?-MHC) genes were analyzed by RT-PCR. RESULTS: Stem cells derived from human placenta were self-renewed and differentiated into cardiomyocytes in vitro. RT-PCR showed that Anf and ?-MHC genes were expressed in the “beating cells”.CONCLUSION: Human placenta is an abundant source of stem cells.
2.Application of Next-Generation Sequencing in Detection of Mutation Gene in ZMPSTE24 in Prenatal Diagnosisa Chinese Pedigree with Pathological Chorioamniotic Membrane Separation
Mingqin MAI ; Yunan WANG ; Xin ZHAO ; Hongke DING ; Jian LU ; Aihua YIN ; Yan CHEN ; Wen WANG ; Hui YI
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(3):453-458
[Objective] To describe a case of a rare,novel mutation causing recurrent chorioamniotic membrane separation in a Chinese family with combined next-generation sequencing (NGS) and Sanger sequencing.[Methods] For the affected fetus,potential mutation were detected by the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing.And the prenatal diagnosis were identified by Sanger sequencing.[Results] A frameshifting mutation c.1389_1390delAG (inherited from mother),and a missense mutationc.1006 G > C (inherited from mother) have been identified in the affected fetus (the second pregnancy).The prenatal diagnosis of the third fetus turns out to be a carrier,the mutation was inherited from father.[Conclusions] We describe a novel mutation in gene ZMPSTE24,which was considered with mandibuloacral dysplasia with type B,and that may be the cousecoursecausing of recurrent chorioamniotic membrane separation.This rare mutation constitutes an additional heterogeneous defect causing chorioamniotic membrane separation.And the conbinedcombined next-generation sequencing (NGS) and Sanger sequencing allows high resolution characterization of novel mutions that are not readily detected by present methods.
3.Fecal-associated microbiome differences between phlegm-dampness constitution and balanced constitution
Li YINI ; Zhao PENGFEI ; Zhang YUNAN ; Zhen JIANHUA ; Zhao LU ; Cai YANAN ; Lu QINGYI ; Huang GUANGRUI
Journal of Traditional Chinese Medical Sciences 2022;9(3):257-266
Objective:This study aimed to explore the structural and functional characteristics of the fecal-associated microbiome(FAM)between the phlegm-dampness constitution(PDC)and balanced consti-tution(BC),and to screen the related specific operational taxonomic unit(OTU)biomarkers.Methods:This was a cross-sectional study.After strictly identifying the constitution of subjects,their clinical index was recorded and counted.Fecal samples were collected for 16S rDNA sequencing.Alpha diversity,beta diversity,and the relative abundance of dominant bacterial taxa were used to describe the FAM structure,and the Wilcoxon rank-sum test,MetagenomeSeq,and linear discriminant analysis effect size(LEfSe)were used to screen specific bacterial taxa.Specific OTUs were screened to construct receiver operating characteristic(ROC)curves.Results:Thirty-two subjects were enrolled,including 22 subjects with BC and 10 subjects with PDC.There were significant differences in cold preference,levels of aspartate transaminase,β2-microglobulin,and creatine kinase MB,and alpha diversity indices(Shannon and Shannoneven)between the two groups.In principal coordinate analysis by abund-jaccard distance measure and partial least squares discriminant analysis,bacterial communities clustered separately between the two groups.Furthermore,based on MetagenomeSeq,LEfSe,and the Wilcoxon rank-sum test,a total of 43,18,and 130 OTUs were differentially distributed between BC group and PDC group,respectively,and OTU200,OTU133,and OTU353 were screened when P<.01.The area under the ROC curve constructed from the 3 selected OTUs was 0.93.Conclusion:The FAM structure and related functional characteristics of the PDC group differed from those of the BC group.In particular,OTU200,OTU133,and OTU353 can be used as unique markers of PDC to assist clinical diagnosis.
4.Optimization of prescription process of lifitegrast eye drops and evaluation of its efficacy for dry eye disease
Xi YAN ; Yunan PAN ; Kaili WANG ; Qianzi XU ; Yan SHEN ; Yang LU
Journal of China Pharmaceutical University 2024;55(2):209-216
In order to develop a more effective drug for dry eye disease, the preparation of lifitegrast eye drops was carried out, and the safety and efficacy of lifitegrast eye drops in vitro and in vivo were investigated. First the method for the determination of lifitegrast content was established, and then the composition and preparation process of the preparation were determined by literature review and single factor experiment. Finally, the safety of lifitegrast eye drops was evaluated by Draize eye irritation test and HE staining, and the therapeutic efficacy was evaluated by Schirmer test and ELISA test. The results showed that the final prescription of lifitegrast eye drops consisted of 5% lifitegrast, 0.4% sodium chloride, 0.3%−0.4% anhydrous disodium hydrogen phosphate, 0.3% sodium thiosulfate pentahydrate and 0.3% sodium hydroxide. The appearance of lifitegrast eye drops was transparent and slightly brownish yellow solution, the pH was7.75±0.05, the osmotic pressure was in the range of 200−330 mOsmol/kg and it had good stability at 60℃ for 3 months. There was no significant difference in irritation study compared with normal saline. Schirmer test showed that tear secretion was increased and the expression of inflammatory factors IL-6, IL-1β and TNF-α in tears were significantly decreased after treatment with lifitegrast eye drops and compared to the commercially available emulsion cyclosporine eye drops, it takes effect faster. The above results indicate that lifitegrast eye drops are simple to prepare and stable, which is a better choice for the rapid treatment of dry eye disease.
5.Study on the inhibitory effect of UBE2T on radiosensitivity of lung adenocarcinoma
Mengjia WU ; Yunan WANG ; Bo HE ; Yanyi LU ; Junzhu XU ; Zixuan SU ; Fengmin YIN ; Shujun LIU ; Yuju BAI ; Wei HU
Chinese Journal of Radiation Oncology 2023;32(6):519-525
Objective:To investigate the effect of ubiquitin binding enzyme 2T (UBE2T) on the radiosensitivity of lung adenocarcinoma and unravel its possible mechanism.Methods:A total of 45 patients pathologically diagnosed with different stages of lung adenocarcinoma and treated with radiotherapy in the Second Affiliated Hospital of Zunyi Medical University from March, 2019 to December, 2021 were enrolled, and the efficacy was evaluated according to response evaluation criteria in solid tumors (RECIST1.1). All patients were divided into radiosensitive group ( n=25) and radioresistant group ( n=20). Radiosensitive group was complete remission (CR)+partial remission (PR), and radioresistant group was stable disease (SD) + progression disease (PD). Immunohistochemistry (IHC) was used to calculate the score based on the staining intensity and the number of positive cells. Chi-square test was combined to analyze the correlation between the expression level of UBE2T in paraffin specimens of lung adenocarcinoma patients and the radiosensitivity of patients. Lentivirus UBE2T-interfered (UBE2Tsh) A549 and UBE2T-overexpressed SPC-A-1 lung adenocarcinoma cells and their respective controls were constructed for irradiation and colony formation assay. The survivor fraction curve was fitted by single-hit multi-target model. The DNA double-strand break (DSB) marker γH2AX foci were detected by immunofluorescence (IF). The expression levels of UBE2T, γH 2AX and Rad51 proteins were detected by Western blot. Cell cycle and apoptosis rate of A549 were determined by flow cytometry. Binary variables were statistically analyzed by Fisher's exact probability method and measurement data were assessed by t-test. Results:High-expression level of UBE2T was correlated with the radiosensitivity of lung adenocarcinoma patients ( P<0.05). UBE2Tsh improved the radiosensitivity of A549 lung adenocarcinoma cells, and the sensitizing enhancement ratio (SER) was 1.795. UBE2T overexpression decreased the radiosensitivity of SPC-A-1 lung adenocarcinoma cells with an SER of 0.293. γH2AX foci number per cell were significantly increased in UBE2Tsh A549 cells after irradiation ( P<0.01) . Compared with the control group, the expression level of γH2AX protein was up-regulated ( P<0.01)and that of Rad51 protein was down-regulated in UBE2Tsh A549 cells after radiation ( P<0.001). Compared with the control group, the expression level of γH2AX protein was down-regulated ( P<0.05) and that of Rad51 protein was up-regulated in UBE2T overexpressed SPC-A-1 cells ( P<0.001). The proportion of UBE2Tsh A549 cells in G 2 phase was decreased ( P<0.01) and cell apoptosis was increased ( P<0.001). Conclusions:UBE2T might promote the radioresistance of lung adenocarcinoma cells by enhancing DNA DSB repair induced by radiotherapy, inducing cell cycle G 2 phase arrest, and reducing cell apoptosis.
6.Application of diversified teaching modes combined with individualized teaching methods in muscle and bone ultrasound training classes
Hongping SONG ; Jing WANG ; Yunan JIA ; Lu LUO ; Jikun HAO ; Dingzhang CHEN
Chinese Journal of Medical Education Research 2024;23(11):1500-1505
Objective:To explore the teaching effect of combining diversified teaching modes with individualized teaching methods in musculoskeletal ultrasound training classes compared with traditional teaching modes.Methods:The 109 trainees participated in the muscle and bone ultrasound training classes from June 2020 to June 2022 were included as the research subjects and divided into the observation group ( n=53) and control group ( n=56). The control group received traditional teaching modes, while the observation group received diversified teaching modes in combination with individualized teaching methods. The differences in teaching satisfaction and final exam scores between the two groups were compared after training. Statistical analysis was conducted using the software SPSS 27.0. Categorical data were presented as cases and percentages, while continuous data were presented as mean±standard deviation. The intergroup differences in continuous variables were compared using the t-test, while the intergroup differences in categorical variables were compared using the chi-square test. Results:There were no significant differences in age, sex, education, and professional title between the control group and the observation group ( P>0.05). The comprehensive satisfaction score of ultrasound teaching was (88.08±6.65) in the observation group and (80.71±8.59) in the control group, with a significant difference between the two groups ( χ2=5.02, P<0.01). Before training, the test scores of the control group and the observation group were (62.61±5.39) and (63.87±7.77), respectively, and there was no significant difference between the two groups ( t=0.98, P=0.330). The final scores, theoretical exam, computer operation, and video reading exam scores of the control group were (84.07±1.49), (82.39±1.97), (85.13±2.55), and (86.79±6.06), respectively, while those of the observation group were (90.06±2.17), (88.28±3.19), (88.92±3.23), and (96.23±5.96); there were significant differences between the two groups in these four indicators ( t=16.75, 11.53, 6.79, and 8.20, respectively; P<0.01). The scores were compared for the two groups of trainees classified based on their unit level, professional title, and educational background. Trainees with the same unit level, professional title, and educational background showed no significant differences in test scores between the two groups, though their final scores were significantly different ( P<0.01). Conclusions:The combination of diversified teaching modes and individualized teaching methods can increase the learning enthusiasm of trainees, improve the teaching effect of training classes, and increase the satisfaction of trainees.
7.Study on calculation method of internal and external fluid capacity of human cells based on bioimpedance spectroscopy.
Li KE ; Jiawei XU ; Yunan ZHAO ; Qiang DU ; Haiming LU
Journal of Biomedical Engineering 2019;36(4):643-648
Bioelectrical impedance measurement technology is a non-invasive detection technology for extracting human physiological and pathological information. The analysis method of the relationship between bioimpedance and human physiological parameters is an important part of this technology. In order to calculate the internal and external liquid volume of human cells more accurately, based on the Moissl equation for calculating the internal and external fluid volume of human cells, a segmented human bioimpedance spectrum measurement model and an improved calculation method of intracellular and external fluid capacity were proposed. The measurement and calculation experiments of the intracellular and extracellular fluid volume before and after the human body's water intake were designed and compared with the Moissl calculation method. The results show that the improved calculation method can calculate the intracellular and extracellular fluid volumes more effectively, and the relative error is less than 5%, which may provide new ideas or more accurate methods for the analysis of human body components, facilitating the diagnosis and treatment of diseases.
Body Water
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Electric Impedance
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Extracellular Fluid
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Humans
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Intracellular Fluid
8.A new pediatric femoral neck system for pediatric femoral neck fractures with a free fracture fragment: a biomechanical analysis
Jinchen CHEN ; Dianhua HUANG ; Yunan LU ; Tianlai CHEN ; Yuwei NING ; Shunyou CHEN
Chinese Journal of Orthopaedic Trauma 2023;25(12):1074-1078
Objective:To investigate the biomechanical properties of a new pediatric femoral neck system in the fixation of pediatric femoral neck fractures with a free fracture fragment.Methods:Ten Sawbones model bones were randomly divided into 2 even groups ( n=5), all of which were made into Delbet Type Ⅱ femoral neck fractures with a Pauwels angle of 70°. A free bone block was removed from the bottom at the proximal end of the fracture to simulate a femoral neck fracture with a free fragment. Group A were fixed with traditional inverted triangle cannulated screws, and group B with a new pediatric femoral neck system. After the 2 groups of specimens were placed on a biomechanical testing machine, each specimen was subjected to a static axial compression test, an anti-torsion test and a cyclic load test in turn. The biomechanical results were compared between groups A and B in aspects of axial compression stiffness, torsional stiffness and maximum displacement difference. Results:The axial compression stiffness [(321.718±5.770) N/mm] and torsional rigidity [(1.448±0.079) N·m/°] in group B were significantly higher than those in group A [(266.722±4.788) N/mm and (1.282±0.023) N·m/°] ( P<0.05). The maximum displacement difference in the cyclic load test in group B [(0.063±0.038) mm] was also significantly smaller than that in group A [(0.117±0.056) mm] ( P<0.05). Conclusion:In fixation of pediatric femoral neck fractures with a free fracture fragment, the new pediatric femoral neck system can lead to better biomechanical stability than the traditional inverted triangle cannulated screws.
9.Comparison of Kirschner wire and clavicular hook plate in the treatment of distal clavicle fractures in older children
Ji LIN ; Yuancheng PAN ; Ran LIN ; Yunan LU ; Hongzhe HUANG ; Jinglin LAI ; Yiwen WANG ; Shunyou CHEN
Chinese Journal of Orthopaedic Trauma 2024;26(8):685-690
Objective:To compare the efficacy between Kirschner wire and clavicular hook plate in the fixation of distal clavicle fractures in older children.Methods:A retrospective analysis was conducted of the 28 pediatric patients with distal clavicle fracture who had undergone surgical treatment at Department of Pediatric Orthopedics, The Second General Hospital of Fuzhou from December 2014 to December 2023. There were 20 boys and 8 girls. Age: 10.0 (9.0, 12.8) years old; 22 left sides and 6 right sides; by the Craig modified Neer classification: 22 cases of type Ⅱ, 5 cases of type Ⅳ, and 1 case of type Ⅴ. The patients were divided into 2 groups based on their internal fixation methods. The Kirschner wire group of 12 cases were treated with open reduction and Kirschner wire fixation; the hook plate group of 16 cases were treated with open reduction and clavicular hook plate fixation. Operation time, hospital stay, fracture healing time, incidence of complications, and scores at the final follow-up (the Constant-Murley shoulder function score, quick disabilities of the arm, shoulder, and hand (quickDASH) score, and visual analog scale (VAS) for aesthetic satisfaction) were recorded and compared between the 2 groups.Results:There were no statistically significant differences in the baseline data between the 2 groups, indicating comparability ( P>0.05). The postoperative follow-up period for the 28 patients was (14.8±6.5) months. The operation time in the Kirschner wire group [(34.3±12.1) min] was significantly shorter than that in the hook plate group [(49.4±16.8) min] ( P<0.05), and the VAS score for aesthetic satisfaction at the final follow-up in the Kirschner wire group [(8.2±1.1) points] was significantly higher than that in the hook plate group [(6.2±2.6) points] ( P<0.05). There were no statistically significant differences between the 2 groups in terms of fracture healing time, hospital stay, incidence of complications, or Constant-Murley score and quickDASH score at the final follow-up ( P>0.05). Conclusions:For pediatric patients with distal clavicle fracture, both Kirschner wire fixation and clavicular hook plate fixation can achieve good functional outcomes. However, Kirschner wire fixation has advantages of shorter operation time, higher postoperative aesthetic satisfaction, and no need of a secondary surgery for implant removal.
10.Mechanism of Shaoyaotang in Treatment of Ulcerative Colitis Based on Network Pharmacology and Experimental Verification
Yini LI ; Yunan ZHANG ; Lu ZHAO ; Lulu WANG ; Yali ZHOU ; Jianhua ZHEN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(23):8-15
ObjectiveTo explore the mechanism of Shaoyaotang (SYT) in the treatment of ulcerative colitis (UC) based on network pharmacology and experimental verification. MethodThe core components, target genes, and main pathways of SYT were predicted based on network pharmacology, and UC-related components, target genes, and pathways were screened. Dextran sodium sulfate (DSS) was used to induce the UC model in mice, and the effect of SYT on UC mice was observed, followed by mechanism verification. ResultNetwork pharmacology indicated that 174 active components and corresponding 159 target genes of SYT were screened, and the related pathways were those mediated by 5-hydroxytryptamine (5-HT) degredation and 5-HT receptor 3. The results of animal experiments showed that compared with the model group, the SYT group showed increased body weight and colon length(P<0.01), reduced disease activity index (DAI) score (P<0.01), improved histopathological manifestations, reduced concentrations of 5-HT in the colonic tissues and serum (P<0.05, P<0.01), and increased mRNA expression of monoamine oxidase A (MAOA), monoamine oxidase B (MAOB), sodium-dependent serotonin transporter (SLC6A4), and 5-HT receptor 3A (5-HTR3A) related to 5-HT metabolism in the colon (P<0.01). ConclusionSYT can alleviate the local inflammatory response of the intestinal tract in UC by regulating 5-HT degredation pathways.