1.Preliminary study on the biological role of EF-hand domain-containing protein 2 in hepatocellular carcinoma
Yanmei ZHANG ; Xiao LI ; Xueqiang JIA ; Juanzi LIU ; Wanqing LI ; Junfeng XUAN ; Shiyu FENG ; Zhaohui SUN ; Weiyun ZHANG
Chinese Journal of Preventive Medicine 2025;59(8):1224-1231
This study investigates the expression pattern and functional significance of EF-hand domain-containing protein 2 (EFHD2) in hepatocellular carcinoma (HCC), with particular focus on its regulatory effects on tumor proliferation, migration, and invasion. Cellular experimental study was completed from June 2024 to January 2025 in the Basic Laboratory of the General Hospital of Southern Theater Command. TCGA database to determine EFHD2 expression and its clinicopathological correlations. GSCA database to assess methylation patterns and immune infiltration. Model of transient overexpression and knockdown of EFHD2 was constructed in hepatocellular carcinoma cells Hep3B, then RT-qPCR and Western blot were applied to verify the transfection efficiency. CCK-8 and colony formation assays for proliferation assessment, Transwell chambers for migration/invasion quantification. Protein-protein interaction networks were constructed via STRING, followed by GO/KEGG enrichment analysis. Statistical analysis was performed using the two independent samples t-test. The results showed that EFHD2 demonstrated significant upregulation in HCC tissues versus normal controls ( P<0.05). Elevated EFHD2 expression correlated with advanced clinical stage ( P<0.05) and poor differentiation ( P<0.05). In the CCK-8 assay, the EFHD2 overexpression group demonstrated significantly higher cell viability than the control group, as evidenced by 450 nm relative absorbance values on Day 1 (0.529±0.019 vs. 0.515±0.016, F=0.041, P=0.320), Day 2 (1.356±0.019 vs. 1.094±0.042, F=3.833, P<0.001), Day 3 (2.817±0.049 vs. 2.143±0.124, F=3.833, P<0.001), and Day 4 (3.848±0.015 vs. 3.430±0.021, F=0.469, P<0.001). The EFHD2 knockdown group showed reduced cell viability compared to controls: Day 1 (0.541±0.020 vs. 0.552±0.015, F=0.098, P=0.423), Day 2 (1.154±0.009 vs. 1.326±0.029, F=2.485, P<0.001), Day 3 (2.453±0.041 vs. 2.653±0.031, F=0.479, P<0.001), and Day 4 (3.685±0.038 vs. 3.836±0.021, F=6.804, P<0.001). In colony formation assays, the overexpression group displayed a significant increase in colony numbers (254.667±23.861 vs. 186.000±16.703, F=0.865, P=0.015), whereas the knockdown group exhibited decreased colony formation (229.000±24.637 vs. 306.667±36.501, F=0.988, P=0.038). In Transwell assays, the EFHD2 overexpression group revealed enhanced migratory capacity [ (605.000±72.670) cells vs. (472.667±28.095) cells, F=2.462, P=0.042] and invasive potential [(767.333±21.221) cells vs. (414.333±16.623) cells, F=0.331, P<0.001]. The knockdown group showed attenuated migration [(311.000±71.084) cells vs. (479.667±50.846) cells, F=0.718, P=0.029] and invasion [(247.667±48.263) cells vs. (345.667±32.130) cells, F=0.727, P=0.043] compared to controls. The network of EFHD2-interacting proteins was further constructed by the STRING database, and the GO and KEGG analysis were used to perform bioinformatics analysis reveal that EFHD2 is mainly involved in actin cytoskeleton regulation. In conclusion, EFHD2 is highly expressed in HCC and is involved in the process of proliferation, migration and invasion of HCC.
2.Effects of MOTOmed intelligent training combined with standing balance training on recovery of lower limb motor function and balance in patients with post-stroke hemiplegia
Jia SUN ; Haiye SHEN ; Shiyu WANG
Journal of Navy Medicine 2025;46(10):1006-1011
Objective To explore the effects of MOTOmed intelligent training combined with standing balance training on the recovery of lower limb motor function and balance in stroke patients with hemiplegia.Methods A total of 124 patients with post-stroke hemiplegia who received rehabilitation care in the Sixth People's Hospital of Nantong from May 2022 to May 2024 were selected as research objects.They were assigned to observation group(62 cases)or control group(62 cases)using a random number table.The control group received MOTOmed intelligent training,and the observation group received MOTOmed intelligent training combined with standing balance training.After 2 months of rehabilitation,the lower limb motor function(Fugl-Meyer assessment lower extremity subscale[FMA-LE]score,time up and go test[TUGT],balance function,postural stability[length and area of center of gravity movement,average trajectory error,completion time],quality of life(stroke specific quality of life[SS-QOL]),and self-care ability(Barthel index[BI])were compared between the two groups.Results After interventions,the FMA-LE scores of the observation group and control group were 19.41±5.72 and 16.08±4.41,respectively;the TUGT result was(40.06±12.35)s and(45.13±10.97)s;the station balance scores were 2.83±0.71 and 2.57±0.63;the walking function balance scores were 2.91±0.43 and 2.75±0.41;the lengths of center of gravity movement were(246.05±45.63)mm and(279.61±42.01)mm;the area of center of gravity movement was(92.75±22.16)mm2 and(102.36±27.84)mm2;the average trajectory error was(21.14±5.06)%and(25.78±5.42)%;the completion time of the tests was(79.24±5.06)s and(82.87±6.73)s;the SS-QOL scores were 138.91±32.05 and 125.65±34.83;BI was 51.24±7.91 and 46.05±6.63.There were significant differences in these indexes between the two groups(all P<0.05).Conclusion MOTOmed intelligent training combined with standing balance training can promote the recovery of lower limb motor function,improve lower limb motor balance,and enhance the quality of life of stroke patients with hemiplegia.
3.Advances in Nanozyme-Aptamer Systems for the Detection of Foodborne Pathogens
Hao LIANG ; Shiyu JIA ; Zhou ZHAN ; Yujiao CAI ; Xiangheng NIU
Journal of Sichuan University (Medical Sciences) 2025;56(5):1251-1259
Food safety problems caused by foodborne pathogenic bacteria pose a serious threat to public health,creating an urgent need to develop testing methods and techniques with excellent performance and are simple to use and of affordable cost.Traditional testing methods,such as isolation and culture,morphological observation,biochemical identification,and serological tests,have many limitations,including complex procedures,reliance on specialized technical equipment and personnel,and long turnaround time,rendering them inadequate for meeting current and future testing demands.Therefore,it is particularly important to develop simple,rapid,and highly sensitive methods for analyzing pathogenic bacteria.The fusion of nucleic acid aptamers and nanozymes brings new ideas for the rapid testing of pathogenic bacteria.On one hand,aptamers offer specific recognition capability for target bacteria and can be combined with various nucleic acid signal amplification techniques.On the other hand,the enzyme-like catalytic activity and signal amplification effect of many nanomaterials provide a basis for highly sensitive testing.This review highlights the application potential of nanozyme?aptamer coupling systems in the field of microbial analysis by briefly summarizing the latest research progress in the use of nanozymes combined with aptamers for the detection of foodborne pathogenic bacteria.First of all,two main approaches to conjugating nanozymes with aptamers are introduced.Then,the testing mechanisms and typical applications of nanozyme?aptamer coupling systems for foodborne pathogenic bacteria are discussed.Finally,future development trends and existing challenges are disucssed from four perspectives,including specificity,high sensitivity,high throughput,and intelligent detection.This review aims to provide a useful reference for the fusion of nanozymes and aptamers and for the development of on-site rapid testing techniques for foodborne pathogens,and to encourage broader academic interest to further advance this promising research field.
4.Screening and identification of African swine fever virus M1249L interacting fac-tors based on yeast two-hybrid system
Shuai CUI ; Yang WANG ; Shiyu CHEN ; Yajun JIANG ; Lichun FANG ; Zhongbao PANG ; Xiaoyu GUO ; Hong JIA ; Hongfei ZHU
Chinese Journal of Veterinary Science 2025;45(11):2301-2308
To explore the interaction between ASFV capsid protein M1249L and host from the host cellular perspective,M1249L was selected for constructing the bait plasmid(pGBKT7-M1249L)to screen the bone marrow-derived macrophages(BMDMs)cDNA library.After again co-transform and sequence alignment,20 candidate interacting host proteins were screened,such as IL-1β,CTSB and DNAJA3.And then,co-immunoprecipitation assay was performed to verify the interaction be-tween M1249L and host proteins.GO ontology(GO)and KEGG pathway enrichment analyses re-vealed that biological regulation,cellular communication and response to stimulus and others were enriched in biological processes.And these host proteins could share some pathways,including toll-like receptor signaling pathway and Nod-like receptor signaling pathway.Therefore,the results provides the theoretical basis for further research on the mechanism of ASFV M1249L in viral in-fection and immune regulation.
5.Preliminary study on the biological role of EF-hand domain-containing protein 2 in hepatocellular carcinoma
Yanmei ZHANG ; Xiao LI ; Xueqiang JIA ; Juanzi LIU ; Wanqing LI ; Junfeng XUAN ; Shiyu FENG ; Zhaohui SUN ; Weiyun ZHANG
Chinese Journal of Preventive Medicine 2025;59(8):1224-1231
This study investigates the expression pattern and functional significance of EF-hand domain-containing protein 2 (EFHD2) in hepatocellular carcinoma (HCC), with particular focus on its regulatory effects on tumor proliferation, migration, and invasion. Cellular experimental study was completed from June 2024 to January 2025 in the Basic Laboratory of the General Hospital of Southern Theater Command. TCGA database to determine EFHD2 expression and its clinicopathological correlations. GSCA database to assess methylation patterns and immune infiltration. Model of transient overexpression and knockdown of EFHD2 was constructed in hepatocellular carcinoma cells Hep3B, then RT-qPCR and Western blot were applied to verify the transfection efficiency. CCK-8 and colony formation assays for proliferation assessment, Transwell chambers for migration/invasion quantification. Protein-protein interaction networks were constructed via STRING, followed by GO/KEGG enrichment analysis. Statistical analysis was performed using the two independent samples t-test. The results showed that EFHD2 demonstrated significant upregulation in HCC tissues versus normal controls ( P<0.05). Elevated EFHD2 expression correlated with advanced clinical stage ( P<0.05) and poor differentiation ( P<0.05). In the CCK-8 assay, the EFHD2 overexpression group demonstrated significantly higher cell viability than the control group, as evidenced by 450 nm relative absorbance values on Day 1 (0.529±0.019 vs. 0.515±0.016, F=0.041, P=0.320), Day 2 (1.356±0.019 vs. 1.094±0.042, F=3.833, P<0.001), Day 3 (2.817±0.049 vs. 2.143±0.124, F=3.833, P<0.001), and Day 4 (3.848±0.015 vs. 3.430±0.021, F=0.469, P<0.001). The EFHD2 knockdown group showed reduced cell viability compared to controls: Day 1 (0.541±0.020 vs. 0.552±0.015, F=0.098, P=0.423), Day 2 (1.154±0.009 vs. 1.326±0.029, F=2.485, P<0.001), Day 3 (2.453±0.041 vs. 2.653±0.031, F=0.479, P<0.001), and Day 4 (3.685±0.038 vs. 3.836±0.021, F=6.804, P<0.001). In colony formation assays, the overexpression group displayed a significant increase in colony numbers (254.667±23.861 vs. 186.000±16.703, F=0.865, P=0.015), whereas the knockdown group exhibited decreased colony formation (229.000±24.637 vs. 306.667±36.501, F=0.988, P=0.038). In Transwell assays, the EFHD2 overexpression group revealed enhanced migratory capacity [ (605.000±72.670) cells vs. (472.667±28.095) cells, F=2.462, P=0.042] and invasive potential [(767.333±21.221) cells vs. (414.333±16.623) cells, F=0.331, P<0.001]. The knockdown group showed attenuated migration [(311.000±71.084) cells vs. (479.667±50.846) cells, F=0.718, P=0.029] and invasion [(247.667±48.263) cells vs. (345.667±32.130) cells, F=0.727, P=0.043] compared to controls. The network of EFHD2-interacting proteins was further constructed by the STRING database, and the GO and KEGG analysis were used to perform bioinformatics analysis reveal that EFHD2 is mainly involved in actin cytoskeleton regulation. In conclusion, EFHD2 is highly expressed in HCC and is involved in the process of proliferation, migration and invasion of HCC.
6.Screening and identification of African swine fever virus M1249L interacting fac-tors based on yeast two-hybrid system
Shuai CUI ; Yang WANG ; Shiyu CHEN ; Yajun JIANG ; Lichun FANG ; Zhongbao PANG ; Xiaoyu GUO ; Hong JIA ; Hongfei ZHU
Chinese Journal of Veterinary Science 2025;45(11):2301-2308
To explore the interaction between ASFV capsid protein M1249L and host from the host cellular perspective,M1249L was selected for constructing the bait plasmid(pGBKT7-M1249L)to screen the bone marrow-derived macrophages(BMDMs)cDNA library.After again co-transform and sequence alignment,20 candidate interacting host proteins were screened,such as IL-1β,CTSB and DNAJA3.And then,co-immunoprecipitation assay was performed to verify the interaction be-tween M1249L and host proteins.GO ontology(GO)and KEGG pathway enrichment analyses re-vealed that biological regulation,cellular communication and response to stimulus and others were enriched in biological processes.And these host proteins could share some pathways,including toll-like receptor signaling pathway and Nod-like receptor signaling pathway.Therefore,the results provides the theoretical basis for further research on the mechanism of ASFV M1249L in viral in-fection and immune regulation.
7.Protocol for the guidelines for the diagnosis and treatment of obesity/overweight in infertile populations (2025)
Jia HUANG ; Linlin JIANG ; Hui CHEN ; Shiyu BAI ; Jing TANG ; Zheng WANG ; Rong LI ; Dongzi YANG
Chinese Journal of Reproduction and Contraception 2024;44(11):1116-1120
Obesity contributes to reduced fertility, infertility and adverse maternal and infant outcomes through multiple mechanisms. Individuals with obesity encounter numerous impediments during fertility treatments and subsequent pregnancy, leading to intensified physical, psychological, and socio-economic burdens. Given the recent progress in research and the expansion of treatment modalities, there is an imperative need to refine the diagnostic and therapeutic frameworks for obese patients with infertility through a multidisciplinary approach. The Reproductive Medicine Specialized Committee of the Chinese Medical Doctor Association (CMDA) has established a multidisciplinary team of experts to develop an evidence-based guideline. This guideline's development will adhere to both national and international standards for guideline development and reporting. This proposal mainly describes the significance and purpose of guideline development, the method of evidence retrieval and quality assessment, the process of guideline development, and the plan for publication, implementation and dissemination of the guideline.
8.Protocol for the guidelines for the diagnosis and treatment of obesity/overweight in infertile populations (2025)
Jia HUANG ; Linlin JIANG ; Hui CHEN ; Shiyu BAI ; Jing TANG ; Zheng WANG ; Rong LI ; Dongzi YANG
Chinese Journal of Reproduction and Contraception 2024;44(11):1116-1120
Obesity contributes to reduced fertility, infertility and adverse maternal and infant outcomes through multiple mechanisms. Individuals with obesity encounter numerous impediments during fertility treatments and subsequent pregnancy, leading to intensified physical, psychological, and socio-economic burdens. Given the recent progress in research and the expansion of treatment modalities, there is an imperative need to refine the diagnostic and therapeutic frameworks for obese patients with infertility through a multidisciplinary approach. The Reproductive Medicine Specialized Committee of the Chinese Medical Doctor Association (CMDA) has established a multidisciplinary team of experts to develop an evidence-based guideline. This guideline's development will adhere to both national and international standards for guideline development and reporting. This proposal mainly describes the significance and purpose of guideline development, the method of evidence retrieval and quality assessment, the process of guideline development, and the plan for publication, implementation and dissemination of the guideline.
9.A biomimetic liver cancer on-a-chip reveals a critical role of LIPOCALIN-2 in promoting hepatocellular carcinoma progression.
Peiliang SHEN ; Yuanyuan JIA ; Weijia ZHOU ; Weiwei ZHENG ; Yueyao WU ; Suchen QU ; Shiyu DU ; Siliang WANG ; Huilian SHI ; Jia SUN ; Xin HAN
Acta Pharmaceutica Sinica B 2023;13(11):4621-4637
Hepatic stellate cells (HSCs) represent a significant component of hepatocellular carcinoma (HCC) microenvironments which play a critical role in tumor progression and drug resistance. Tumor-on-a-chip technology has provided a powerful in vitro platform to investigate the crosstalk between activated HSCs and HCC cells by mimicking physiological architecture with precise spatiotemporal control. Here we developed a tri-cell culture microfluidic chip to evaluate the impact of HSCs on HCC progression. On-chip analysis revealed activated HSCs contributed to endothelial invasion, HCC drug resistance and natural killer (NK) cell exhaustion. Cytokine array and RNA sequencing analysis were combined to indicate the iron-binding protein LIPOCALIN-2 (LCN-2) as a key factor in remodeling tumor microenvironments in the HCC-on-a-chip. LCN-2 targeted therapy demonstrated robust anti-tumor effects both in vitro 3D biomimetic chip and in vivo mouse model, including angiogenesis inhibition, sorafenib sensitivity promotion and NK-cell cytotoxicity enhancement. Taken together, the microfluidic platform exhibited obvious advantages in mimicking functional characteristics of tumor microenvironments and developing targeted therapies.
10.Methodological study on in vitro release of etonogestrel/ethinyl estradiol compound vaginal ring
Shasha WANG ; Pan CHEN ; Jia ZENG ; Man XUE ; Fang LI ; Feng PAN ; Yanhong ZHAO ; Shiyu MA ; Fuzheng REN ; Yanxiong KE ; Linglin FENG
Chinese Journal of Reproduction and Contraception 2023;43(8):813-819
Objective:To develop an etonogestrel (ENG)/ethinyl estradiol (EE) compound vaginal ring with good sustained-release properties, studying its in vitro release assay methodology, and establishing an accelerated release method with good correlation with real-time release methods. Methods:Ethylene-vinyl acetate copolymer (EVA) was used as the matrix to prepare ENG/EE compound vaginal ring by hot melt extrusion method, and an in vitro real-time release assay method and an accelerated release assay method based on shake flask method were established. The in vitro release behavior and release mechanism of ENG/EE compound vaginal ring were investigated, and the correlation between real-time release and accelerated release over time was analyzed. Results:The obtained vaginal ring had a round appearance and a smooth surface, and the drug was slowly released under the optimal real-time release conditions in vitro (release medium: 200 mL of pure water, shaking at 60 r/min in a constant temperature water bath shaker at 37 ℃), and the cumulative release of ENG and EE on the 21st day were 54.39% and 46.80%, respectively. The correlation coefficients of drug release and its mechanism under the optimized in vitro accelerated conditions (release medium: 200 mL of 0.6% sodium dodecyl sulfate aqueous solution, shaking at 60 r/min at 55 ℃) with the results under real-time release conditions were all greater than 0.99. Conclusion:The ENG/EE compound vaginal ring obtained in this study has a significant in vitro sustained-release effect, and the established accelerated release conditions have a good correlation with real-time release conditions, which can provide a useful reference for the quality evaluation research of such vaginal ring products and the formulation of guidelines for in vitro release research methods.

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