1.Analysis of human parvovirus B19 nucleic acid detection in blood products in China
Yue WANG ; Xiaobei ZHENG ; Qin GONG ; Ying ZHAO ; Yuanxiu LUO ; Dandan YANG ; Linlin ZHANG ; Zheng JIANG ; Gan PENG ; Jin ZHANG ; Bingbing KE
Chinese Journal of Blood Transfusion 2025;38(7):950-957
Objective: To analyze the nucleic acid load of human parvovirus B19 in major commercially available blood products in China, including human albumin, human intravenous immunoglobulin, human rabies immunoglobulin and various coagulation factor products, aiming to provide evidence for improving blood product manufacturing processes and quality control of source plasma. Methods: A total of 98 batches of coagulation factor products were tested for human parvovirus B19 nucleic acid using real-time fluorescent quantitative PCR, including 42 batches of human prothrombin complex, 35 batches of human coagulation factor Ⅷ, and 21 batches of human fibrinogen. Additionally, 6 batches of human albumin, 6 batches of human intravenous immunoglobulin, and 38 batches of human rabies immunoglobulin were tested for human parvovirus B19 nucleic acid. Results: Human parvovirus B19 nucleic acid were undetectable in human albumin, human intravenous immunoglobulin and human rabies immunoglobulin. Among the 98 batches of coagulation factor products tested for human parvovirus B19 nucleic acid, B19 nucleic acid reactivity rate was 69.0% (29/42) for human prothrombin complex batches, but nucleic acid concentration were all significantly lower than 10
IU/mL. The reactivity rate of B19 nucleic acid in 35 batches of human coagulation factor Ⅷ was 48.6% (17/35), with nucleic acid concentration all below 10
IU/mL. The reactivity rate of B19 nucleic acid in 21 batches of human fibrinogen was 61.9% (13/21), with nucleic acid concentration all below 10
IU/mL. Conclusion: No human parvovirus B19 has been detected in human albumin, human intravenous immunoglobulin, or human rabies immunoglobulin. Human parvovirus B19 nucleic acid may exist in commercially available coagulation factor products, highlighting the need for enhanced screening of human parvovirus B19 nucleic acid in these products. It is also recommended that B19 viral nucleic acid testing be conducted on source plasma, particularly for coagulation factor products.
2.Protective effect of modified University of Wisconsin preservation solution containing α 2-adrenergic receptor agonists and noble gases on isolated amputated skeletal muscle of rats
Zhengwei XUE ; Zhigang QIN ; Xiangfeng LIU ; Jieyu LI ; Ling JIANG ; Xiao LI ; Jianbo MA ; Guanlei LIU ; Pengfei ZHENG ; Ying TANG ; Peng LI ; Jianteng GU
Chinese Journal of Trauma 2025;41(11):1112-1122
Objective:To develop a modified University of Wisconsin preservation solution (UW solution) containing α 2-adrenergic receptor agonists (dexmedetomidine) and noble gases (argon) and investigate its protective effect on the isolated amputated skeletal muscle of rats. Methods:Sixty male SD rats were selected to establish a hindlimb cold preservation/perfusion model and were divided into blank control group, hypothermic storage group, UW solution perfusion group, and modified UW solution perfusion group using a random number table, with 15 rats in each group. Simultaneously, a cold preservation model of rat skeletal muscle myoblasts (L6 cells) was established and the rats were also divided into four groups in the same way. Animal models were prepared in different ways: In the blank control group, the hindlimbs received no special treatment; In the hypothermic storage group, the amputated hindlimbs were stored in a dry centrifuge tube at 4℃ for 18 hours; In the UW solution perfusion group, the amputated hindlimbs were perfused with UW solution and then stored in a centrifuge tube containing UW solution at 4℃ for 18 hours; In the modified UW solution perfusion group, the amputated hindlimbs were perfused with modified UW solution (containing 0.1 nmol/L dexmedetomidine and 50% volume fraction of argon) and then stored in a centrifuge tube containing the modified UW solution at 4℃ for 18 hours. Cell models were treated as follows: In the blank control group, L6 cells were cultured under standard conditions; In the hypothermic storage group and UW solution group, L6 cells were treated with conventional culture medium or UW solution, stored in argon-filled sealed bags at 4℃ for 8 hours, and then rewarmed and cultured for 6 hours; In the modified UW solution group, L6 cells were treated with the modified solution, stored in argon-filled sealed bags at 4℃ for 8 hours, and then rewarmed and cultured for 6 hours. After sample collection, skeletal muscle morphology, tissue edema and ultrastructure features were assessed by HE staining, wet-to-dry weight ratio, and transmission electron microscopy, respectively. Additionally, L6 cell morphology was examined by light microscopy. L6 cell viability was determined by cell counting kit-8 (CCK-8) assay (expressed as absorbance A value). Expression levels of glutathione peroxidase 4 (GPX4) protein in both skeletal muscle tissue and L6 cells were evaluated by immunofluorescence staining and Western blot, respectively.Results:After 18 hours of in vitro preservation of rat isolated amputated limbs, the following results were obtained: (1) HE staining results showed that the muscle fiber morphology of the modified UW solution perfusion group was close to that of the blank control group. Moreover, the area ratio of skeletal muscle cells in the modified UW solution perfusion group was significantly higher than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). (2) The wet-dry weight ratio results showed that there was no statistically significant difference among the modified UW solution perfusion group, the blank control group and UW solution group ( P>0.05), with significantly lower ratios in all three groups than that in the hypothermic storage group ( P<0.05). (3) Transmission electron microscopy results revealed that the modified UW solution perfusion group showed no statistically significant differences in ultrastructural metrics, including myofiber diameter, sarcomere length, mitochondrial short-axis/long-axis ratio, and mitochondrial cristae count, compared with those in the blank control group ( P>0.05), and performed significantly better than both the hypothermic storage group and UW solution perfusion group ( P<0.05). (4) Morphological observation of L6 cells showed that the cellular morphology was regular in the modified UW solution perfusion group, close to that in the blank control group, while it was severely damaged in the hypothermic storage group. Moreover, the cells were reduced in number and partially damaged in the UW solution group. The sequence of cell viability expressed as absorbance A value was blank control group >modified UW solution perfusion group > UW solution perfusion group > hypothermic storage group, with statistically significant differences among the four groups ( P<0.05). (5) Immunofluorescence staining showed that there was no statistically significant difference in fluorescence intensity of GPX4 protein expression between the modified UW solution perfusion group and blank control group ( P>0.05), while the fluorescence intensity was higher in the modified UW solution perfusion group than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). Western blot analysis showed that the relative expression level of GPX4 in the modified UW solution group was significantly lower than that in the blank control group ( P<0.05), but higher than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). Conclusion:The modified UW solution can stabilize the expression level of GPX4 protein, thereby inhibiting ferroptosis and alleviating cold preservation injury in both rat amputated isolated limb skeletal muscle tissue and L6 cells.
3.Study on the Inhibitory Effect of Bruceine A on Wnt/β-catenin Signaling Pathway against Proliferation and Metastasis of Colorectal Cancer Cells
Yuxiang SHENG ; Jie JIANG ; Jiafu YUAN ; Xiaojiang BAO ; Zheng ZHENG ; Fuhao QIN ; Yishi TANG ; Bin JIANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):223-232
OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.
4.Analysis of human parvovirus B19 nucleic acid detection in blood products in China
Yue WANG ; Xiaobei ZHENG ; Qin GONG ; Ying ZHAO ; Yuanxiu LUO ; Dandan YANG ; Linlin ZHANG ; Zheng JIANG ; Gan PENG ; Jin ZHANG ; Bingbing KE
Chinese Journal of Blood Transfusion 2025;38(7):950-957
Objective: To analyze the nucleic acid load of human parvovirus B19 in major commercially available blood products in China, including human albumin, human intravenous immunoglobulin, human rabies immunoglobulin and various coagulation factor products, aiming to provide evidence for improving blood product manufacturing processes and quality control of source plasma. Methods: A total of 98 batches of coagulation factor products were tested for human parvovirus B19 nucleic acid using real-time fluorescent quantitative PCR, including 42 batches of human prothrombin complex, 35 batches of human coagulation factor Ⅷ, and 21 batches of human fibrinogen. Additionally, 6 batches of human albumin, 6 batches of human intravenous immunoglobulin, and 38 batches of human rabies immunoglobulin were tested for human parvovirus B19 nucleic acid. Results: Human parvovirus B19 nucleic acid were undetectable in human albumin, human intravenous immunoglobulin and human rabies immunoglobulin. Among the 98 batches of coagulation factor products tested for human parvovirus B19 nucleic acid, B19 nucleic acid reactivity rate was 69.0% (29/42) for human prothrombin complex batches, but nucleic acid concentration were all significantly lower than 10
IU/mL. The reactivity rate of B19 nucleic acid in 35 batches of human coagulation factor Ⅷ was 48.6% (17/35), with nucleic acid concentration all below 10
IU/mL. The reactivity rate of B19 nucleic acid in 21 batches of human fibrinogen was 61.9% (13/21), with nucleic acid concentration all below 10
IU/mL. Conclusion: No human parvovirus B19 has been detected in human albumin, human intravenous immunoglobulin, or human rabies immunoglobulin. Human parvovirus B19 nucleic acid may exist in commercially available coagulation factor products, highlighting the need for enhanced screening of human parvovirus B19 nucleic acid in these products. It is also recommended that B19 viral nucleic acid testing be conducted on source plasma, particularly for coagulation factor products.
5.Investigation of an outbreak of group A human G9P [8] rotavirus infectious diarrhea among adults in Chongqing
Yang WANG ; Yuan KONG ; Ning CHEN ; Lundi YANG ; Jiang LONG ; Qin LI ; Xiaoyang XU ; Wei ZHENG ; Hong WEI ; Jie LU ; Quanjie XIAO ; Yingying BA ; Wenxi WU ; Qian XU ; Ju YAN
Shanghai Journal of Preventive Medicine 2025;37(8):663-668
ObjectiveTo investigate and analyze an outbreak of rotavirus infectious diarrhea in a prison in Chongqing Municipality, to provide a basis for adult rotavirus surveillance and prevention, and to explore the public health problems in special settings. MethodsA retrospective survey was conducted to collect and analyze data on individual cases with diarrheal disease on-site. The clinical characteristics, as well as the temporal, spatial and geographical distribution patterns of the epidemic were described. Multi-pathogen detection tests were conducted both on diarrhea cases and environmental samples, with viral genotyping performed on positive samples. A case-control analysis was performed to identify the causes of the outbreak, and an SEIR model was adopted to predict the outbreak trend and evaluate the effectiveness of interventions. ResultsA total of 65 cases were found among the inmates, with an attack rate of 2.03%. The predominant clinical manifestations included diarrhea (89.23%), watery stool (73.85%), and dehydration (18.46%). The epidemic curve indicated a “human-to-human” transmission pattern, with an average incubation period of 5‒6 days. The attack rates among chefs in the main canteen (80.00%, 8/10) and caterers (28.33%, 17/60) were significantly higher than those of other inmates (P<0.05). Multi-pathogen polymerase chain reaction (PCR) testing detected positive for group A rotavirus, with the viral genotyping identified as G9P [8] strain. Factors such as unprotected "bare-handed" food distribution among cases with diarrhea (OR=9.512, 95%CI: 4.261‒21.234) and close contact with diarrhea cases (OR=3.656, 95%CI: 1.719‒7.778) were the possible cause of the outbreak. The SEIR model (r0=5, α=0.3, β1=0.08, β2=0.04) was constructed using prison inmates as susceptible population, aiming at fitting the initial transmission trend of the outbreak, and the epidemic rate declined rapidly after intervention measures were implemented (rt≈0). ConclusionThis rare rotavirus infection diarrhea outbreak among adults in confined settings suggests that the construction of public health prevention and control systems in prison may be overlooked. Cross infection during meal processing and distribution in the canteens of such settings is likely to be the cause of the outbreak. Given the potential neglect of public heath system construction in special settings, it is imperative to enhance the surveillance and monitoring of rotavirus and other intestinal multi-pathogens among adults, as well as the construction of public health prevention and control systems in these special settings.
6.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
7.Epidemic characteristics and trend analysis of acute hemorrhagic conjunctivitis in China from 2015 to 2022
Yunfei ZHANG ; Yue SHI ; Jianfeng JIANG ; Xuedong ZHENG ; Baijun JIN ; Zhaorui CHANG ; Tian QIN ; Mengjie GENG ; Shiwen WANG
Chinese Journal of Experimental and Clinical Virology 2025;39(3):270-277
Objective:To analyze the epidemiological characteristics and trend of acute hemorrhagic conjunctivitis (AHC) in China from 2015 to 2022, and to provide evidence for the adjustment of prevention and control strategies and measures for AHC.Methods:The case data of AHC reported by national notifiable disease information system from 2015 to 2022 were collected, and descriptive analysis method were used to analyze the population distribution characteristics, temporal epidemiological trends and spatial clusters of AHC in China.Results:From 2015 to 2022, the incidence of AHC in China ranged from 1.85/100 000 to 2.97/100 000, with a fluctuating downward trend. The average annual percent change (AAPC) was -4.91 (95% CI: -7.74 to -2.00, P<0.05), with an annual percent change (APC) of 2.73 (95% CI: -2.34 to 8.06, P=0.189) for 2015—2019 and an APC of -14.23 (95% CI: -21.78 to -5.94, P<0.05). The age-specific incidence rate was highest in children aged 0-4 years (fluctuating between 4.69 and 5.67/100 000 from 2015 to 2019; It decreased significantly during 2020—2022, fluctuating between 1.93 and 2.72 per 100 000).The proportion of cases in children at 0-4 years of age showed a fluctuating downward trend from 8.68% in 2015 to 3.76% in 2020, with an increase in 2021—2022 to 5.74%. After 2020, the proportion of the population aged 60 years and above has increased, reaching 33.59% in 2022. Cases were mainly farmers, with a fluctuating upward trend of around 50% per year, with the highest percentage of 60.96% in 2020. The peak seasonal incidence of AHC was obvious from May to September from 2015 to 2019, but it was not obvious in 2020—2022. The cases were mainly distributed in Guangxi, Hainan and other provinces in the southwestern part of China. The high incidence counties were concentrated in Leye County of Guangxi, Maojian District of Hubei, Fuchuan Yao Autonomous County of Guangxi, Funing County of Yunnan, and Pulan County of Tibet every year. Conclusions:The overall epidemic rate of AHC in China showed a fluctuating downward trend from 2015 to 2022, with a pronounced decline observed between 2020 and 2022, potentially linked to non-pharmaceutical interventions during the COVID-19 pandemic. Increased attention needs to be given to farmers and people above 60 years of age to reduce the risk of morbidity. Moreover, prevention and control efforts should be strengthened in high-risk areas of southwestern China, and comprehensive measures should be implemented in counties with high incidence, including enhanced health education campaigns and improved allocation of sanitary facilities, to reduce the risk of AHC infection. This study is the first to highlight the potential impact of public health policies on AHC epidemiology, thereby offering a scientific foundation for population- and region-specific precision prevention strategies, particularly guiding the refinement of control measures in high-burden areas.
8.Expert consensus on the standard of practice for modified electro-convulsive therapy for mental disorders
Xiu ZHANG ; Guohui LAO ; Xiong HUANG ; Wei JIANG ; Qingmei KONG ; Wei LI ; Hu DENG ; Jijun WANG ; Qin XIE ; Wei DENG ; Shaohua HU ; Dongsheng ZHOU ; Xin WEI ; Zhanming SHI ; Cuixia AN ; Sha LIU ; Yanghua TIAN ; Decheng ZOU ; Lingyun ZENG ; Kun LI ; Xingbing HUANG ; Wei ZHENG ; Yuping NING
Chinese Journal of Psychiatry 2025;58(7):506-525
As a physical treatment technique, modified electro-convulsive therapy (MECT) is used to treat mental and certain neurological disorders by causing seizures with short, suitable electrical currents applied to the brain while the patient is under general anesthesia and muscle relaxants. MECT is recognized for its therapeutic efficacy and clinical safety, rendering it one of the most prevalent interventions in psychiatric care. To enhance clinical outcomes and minimize adverse effects, this consensus document delineates the indications, therapeutic parameters, therapeutic procedures, potential adverse effects, and associated management strategies for MECT. These guidelines are informed by the latest clinical research and expert consensus, integrating evidence-based medicine methodologies. The objective is to furnish clinicians with precise operational guidelines and to advance the standardization of MECT practices in clinical settings.
9.Study on the Inhibitory Effect of Bruceine A on Wnt/β-catenin Signaling Pathway against Proliferation and Metastasis of Colorectal Cancer Cells
Yuxiang SHENG ; Jie JIANG ; Jiafu YUAN ; Xiaojiang BAO ; Zheng ZHENG ; Fuhao QIN ; Yishi TANG ; Bin JIANG
Journal of Nanjing University of Traditional Chinese Medicine 2025;41(2):223-232
OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.
10.Protective effect of modified University of Wisconsin preservation solution containing α 2-adrenergic receptor agonists and noble gases on isolated amputated skeletal muscle of rats
Zhengwei XUE ; Zhigang QIN ; Xiangfeng LIU ; Jieyu LI ; Ling JIANG ; Xiao LI ; Jianbo MA ; Guanlei LIU ; Pengfei ZHENG ; Ying TANG ; Peng LI ; Jianteng GU
Chinese Journal of Trauma 2025;41(11):1112-1122
Objective:To develop a modified University of Wisconsin preservation solution (UW solution) containing α 2-adrenergic receptor agonists (dexmedetomidine) and noble gases (argon) and investigate its protective effect on the isolated amputated skeletal muscle of rats. Methods:Sixty male SD rats were selected to establish a hindlimb cold preservation/perfusion model and were divided into blank control group, hypothermic storage group, UW solution perfusion group, and modified UW solution perfusion group using a random number table, with 15 rats in each group. Simultaneously, a cold preservation model of rat skeletal muscle myoblasts (L6 cells) was established and the rats were also divided into four groups in the same way. Animal models were prepared in different ways: In the blank control group, the hindlimbs received no special treatment; In the hypothermic storage group, the amputated hindlimbs were stored in a dry centrifuge tube at 4℃ for 18 hours; In the UW solution perfusion group, the amputated hindlimbs were perfused with UW solution and then stored in a centrifuge tube containing UW solution at 4℃ for 18 hours; In the modified UW solution perfusion group, the amputated hindlimbs were perfused with modified UW solution (containing 0.1 nmol/L dexmedetomidine and 50% volume fraction of argon) and then stored in a centrifuge tube containing the modified UW solution at 4℃ for 18 hours. Cell models were treated as follows: In the blank control group, L6 cells were cultured under standard conditions; In the hypothermic storage group and UW solution group, L6 cells were treated with conventional culture medium or UW solution, stored in argon-filled sealed bags at 4℃ for 8 hours, and then rewarmed and cultured for 6 hours; In the modified UW solution group, L6 cells were treated with the modified solution, stored in argon-filled sealed bags at 4℃ for 8 hours, and then rewarmed and cultured for 6 hours. After sample collection, skeletal muscle morphology, tissue edema and ultrastructure features were assessed by HE staining, wet-to-dry weight ratio, and transmission electron microscopy, respectively. Additionally, L6 cell morphology was examined by light microscopy. L6 cell viability was determined by cell counting kit-8 (CCK-8) assay (expressed as absorbance A value). Expression levels of glutathione peroxidase 4 (GPX4) protein in both skeletal muscle tissue and L6 cells were evaluated by immunofluorescence staining and Western blot, respectively.Results:After 18 hours of in vitro preservation of rat isolated amputated limbs, the following results were obtained: (1) HE staining results showed that the muscle fiber morphology of the modified UW solution perfusion group was close to that of the blank control group. Moreover, the area ratio of skeletal muscle cells in the modified UW solution perfusion group was significantly higher than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). (2) The wet-dry weight ratio results showed that there was no statistically significant difference among the modified UW solution perfusion group, the blank control group and UW solution group ( P>0.05), with significantly lower ratios in all three groups than that in the hypothermic storage group ( P<0.05). (3) Transmission electron microscopy results revealed that the modified UW solution perfusion group showed no statistically significant differences in ultrastructural metrics, including myofiber diameter, sarcomere length, mitochondrial short-axis/long-axis ratio, and mitochondrial cristae count, compared with those in the blank control group ( P>0.05), and performed significantly better than both the hypothermic storage group and UW solution perfusion group ( P<0.05). (4) Morphological observation of L6 cells showed that the cellular morphology was regular in the modified UW solution perfusion group, close to that in the blank control group, while it was severely damaged in the hypothermic storage group. Moreover, the cells were reduced in number and partially damaged in the UW solution group. The sequence of cell viability expressed as absorbance A value was blank control group >modified UW solution perfusion group > UW solution perfusion group > hypothermic storage group, with statistically significant differences among the four groups ( P<0.05). (5) Immunofluorescence staining showed that there was no statistically significant difference in fluorescence intensity of GPX4 protein expression between the modified UW solution perfusion group and blank control group ( P>0.05), while the fluorescence intensity was higher in the modified UW solution perfusion group than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). Western blot analysis showed that the relative expression level of GPX4 in the modified UW solution group was significantly lower than that in the blank control group ( P<0.05), but higher than that in the hypothermic storage group and UW solution perfusion group ( P<0.05). Conclusion:The modified UW solution can stabilize the expression level of GPX4 protein, thereby inhibiting ferroptosis and alleviating cold preservation injury in both rat amputated isolated limb skeletal muscle tissue and L6 cells.

Result Analysis
Print
Save
E-mail