1.Research on the mechanism of leptin regulating core binding factor β to promote chondrocyte apoptosis
Jiafei YANG ; Zhujun ZHOU ; Guangdi LI ; Yuan HUANG ; Mi ZHANG ; Lianghong DONG
Chinese Journal of Orthopaedics 2025;45(7):436-445
Objective:To investigate the regulatory effect of leptin via the JAK2/STAT3 pathway on the core-binding factor β-subunit (CBFβ) and its molecular mechanism in promoting chondrocyte apoptosis.Methods:A total of five patients undergoing total knee arthroplasty due to knee osteoarthritis (OA group) and five patients undergoing amputation due to trauma (amputation group) were enrolled, and knee cartilage samples were obtained intraoperatively. Western blotting was used to detect the protein expression levels of leptin, CBFβ, matrix metalloproteinase-1 (MMP1), and MMP13. Flow cytometry was performed to determine the optimal treatment duration and concentration of leptin. Chondrocytes were divided into the following groups based on treatment conditions: control group (untreated chondrocytes), leptin group (chondrocytes treated with 50 ng/ml leptin), negative leptin group (chondrocytes transfected with a non-targeting sequence as a control), and leptin+shCBFβ group (chondrocytes transfected with shCBFβ to inhibit CBFβ expression). Apoptosis and the expression levels of MMP1 and MMP13 were analyzed in the four groups. Additionally, chondrocytes were categorized into the following groups for further analysis: control group (untreated cells), leptin group (cells stimulated with 50 ng/ml leptin for 48 h), AG490 group (cells treated with the JAK2/STAT3 inhibitor AG490), and leptin+AG490 group (cells pretreated with AG490 for 2 h followed by 50 ng/ml leptin stimulation for 48 h). The protein expression levels of CBFβ, MMP1, and MMP13, as well as the apoptosis rate, were examined in the four groups.Results:The relative expression levels of leptin, CBFβ, MMP1, and MMP13 in the amputation group were 0.66±0.06, 0.69±0.06, 0.74±0.05, and 0.41±0.03, respectively, which were significantly lower than those in the OA group (1.04±0.10, 1.06±0.09, 0.95±0.04, and 0.99±0.09, respectively) ( P<0.05). The optimal treatment duration and concentration of leptin were determined to be 48 h and 50 ng/ml, respectively. The expression levels of MMP1 and MMP13 significantly differed among the control, leptin, negative leptin, and leptin+shCBFβ groups ( P<0.05). Specifically, the leptin group showed higher expression levels compared to the control group, while the leptin+shCBFβ group exhibited lower expression levels than the leptin group ( P<0.05). The apoptosis rates of chondrocytes in the four groups were 4.55%±1.30%, 22.52%±2.03%, 22.03%±2.01%, and 5.15%±0.91%, respectively, with significant differences ( F=114.066, P<0.001). The apoptosis rate in the leptin group was significantly higher than that in the control group, while the leptin+shCBFβ group exhibited a significantly lower apoptosis rate than the leptin group ( P<0.05). Similarly, significant differences were observed in the expression levels of CBFβ, MMP1, and MMP13 among the control, leptin, AG490, and leptin+AG490 groups ( P<0.05). The expression levels in the leptin group were higher than those in the control group, while the leptin+AG490 group exhibited lower expression levels compared to the leptin group ( P<0.05). The apoptosis rates of chondrocytes in the control, leptin, AG490, and leptin+AG490 groups were 5.19±0.94%, 31.52±2.63%, 5.51±1.41%, and 10.47±0.85%, respectively, with significant differences ( F=117.104, P<0.001). The apoptosis rate in the leptin group was significantly higher than that in the control group, while the leptin+AG490 group exhibited a significantly lower apoptosis rate than the leptin group ( P<0.05). Conclusion:Leptin promotes CBFβ expression via the JAK2/STAT3 pathway, leading to chondrocyte apoptosis and extracellular matrix degradation.
2.Research on the mechanism of leptin regulating core binding factor β to promote chondrocyte apoptosis
Jiafei YANG ; Zhujun ZHOU ; Guangdi LI ; Yuan HUANG ; Mi ZHANG ; Lianghong DONG
Chinese Journal of Orthopaedics 2025;45(7):436-445
Objective:To investigate the regulatory effect of leptin via the JAK2/STAT3 pathway on the core-binding factor β-subunit (CBFβ) and its molecular mechanism in promoting chondrocyte apoptosis.Methods:A total of five patients undergoing total knee arthroplasty due to knee osteoarthritis (OA group) and five patients undergoing amputation due to trauma (amputation group) were enrolled, and knee cartilage samples were obtained intraoperatively. Western blotting was used to detect the protein expression levels of leptin, CBFβ, matrix metalloproteinase-1 (MMP1), and MMP13. Flow cytometry was performed to determine the optimal treatment duration and concentration of leptin. Chondrocytes were divided into the following groups based on treatment conditions: control group (untreated chondrocytes), leptin group (chondrocytes treated with 50 ng/ml leptin), negative leptin group (chondrocytes transfected with a non-targeting sequence as a control), and leptin+shCBFβ group (chondrocytes transfected with shCBFβ to inhibit CBFβ expression). Apoptosis and the expression levels of MMP1 and MMP13 were analyzed in the four groups. Additionally, chondrocytes were categorized into the following groups for further analysis: control group (untreated cells), leptin group (cells stimulated with 50 ng/ml leptin for 48 h), AG490 group (cells treated with the JAK2/STAT3 inhibitor AG490), and leptin+AG490 group (cells pretreated with AG490 for 2 h followed by 50 ng/ml leptin stimulation for 48 h). The protein expression levels of CBFβ, MMP1, and MMP13, as well as the apoptosis rate, were examined in the four groups.Results:The relative expression levels of leptin, CBFβ, MMP1, and MMP13 in the amputation group were 0.66±0.06, 0.69±0.06, 0.74±0.05, and 0.41±0.03, respectively, which were significantly lower than those in the OA group (1.04±0.10, 1.06±0.09, 0.95±0.04, and 0.99±0.09, respectively) ( P<0.05). The optimal treatment duration and concentration of leptin were determined to be 48 h and 50 ng/ml, respectively. The expression levels of MMP1 and MMP13 significantly differed among the control, leptin, negative leptin, and leptin+shCBFβ groups ( P<0.05). Specifically, the leptin group showed higher expression levels compared to the control group, while the leptin+shCBFβ group exhibited lower expression levels than the leptin group ( P<0.05). The apoptosis rates of chondrocytes in the four groups were 4.55%±1.30%, 22.52%±2.03%, 22.03%±2.01%, and 5.15%±0.91%, respectively, with significant differences ( F=114.066, P<0.001). The apoptosis rate in the leptin group was significantly higher than that in the control group, while the leptin+shCBFβ group exhibited a significantly lower apoptosis rate than the leptin group ( P<0.05). Similarly, significant differences were observed in the expression levels of CBFβ, MMP1, and MMP13 among the control, leptin, AG490, and leptin+AG490 groups ( P<0.05). The expression levels in the leptin group were higher than those in the control group, while the leptin+AG490 group exhibited lower expression levels compared to the leptin group ( P<0.05). The apoptosis rates of chondrocytes in the control, leptin, AG490, and leptin+AG490 groups were 5.19±0.94%, 31.52±2.63%, 5.51±1.41%, and 10.47±0.85%, respectively, with significant differences ( F=117.104, P<0.001). The apoptosis rate in the leptin group was significantly higher than that in the control group, while the leptin+AG490 group exhibited a significantly lower apoptosis rate than the leptin group ( P<0.05). Conclusion:Leptin promotes CBFβ expression via the JAK2/STAT3 pathway, leading to chondrocyte apoptosis and extracellular matrix degradation.
3.Host MYH9 protein promotes Zika virus invasion in U251 cells
Qianyi PENG ; Yalan FENG ; Jing HE ; Rong HUANG ; Jiafei ZHAN ; Yuhang ZHENG ; Chen CHEN ; Rong XIA ; Lei YUAN ; Jian YANG ; Kui XU
Chinese Journal of Experimental and Clinical Virology 2025;39(3):278-286
Objective:This study aimed to investigate the regulatory role and mechanism of myosin heavy chain 9 (MYH9) in the invasion of Zika virus (ZIKV) into human glioma cells (U251).Methods:Utilizing CRISPR/Cas9 technology, MYH9-knockout U251 cells (U251-MYH9 KD) were constructed. Following ZIKV infection, the protein expression levels, RNA load, and viral titer of ZIKV were detected through western blot (WB), Real-time fluorescence quantitative polymerase chain reaction (qPCR), and plaque formation assays, respectively. The infection efficiency of ZIKV in U251 cells treated with the MYH9 inhibitor blebbistatin was assessed. The binding and internalization efficiency of ZIKV were measured in U251-MYH9 KD cells. The interaction between MYH9 and the ZIKV envelope protein (E) was studied using co-immunoprecipitation (Co-IP). The effects of soluble MYH9 recombinant protein and anti-human MYH9 antibodies on ZIKV infection were evaluated by qPCR and plaque formation assays. Results:It was found that knockout or inhibition of MYH9 significantly suppressed ZIKV infection in U251 cells. MYH9 knockout notably inhibited the binding and internalization of ZIKV in U251 cells. MYH9 interacted with the ZIKV E protein, and both MYH9 recombinant protein and anti-human MYH9 antibodies, by blocking the binding of ZIKV E protein to cell surface MYH9, inhibited ZIKV infection in U251 cells in a dose-dependent manner.Conclusions:MYH9 facilitates ZIKV invasion into U251 cells through interaction with the ZIKV E protein.
4.Visual analysis of research hotspots and trends in negative-pressure wound therapy in the field of wounds in the past 5 years
Shiqi WEN ; Liping HUANG ; Jiafei LI ; Yuwen CHEN ; Ka LI
Chinese Journal of Modern Nursing 2025;31(26):3549-3555
Objective:To visualize and analyze the literature on the application of negative-pressure wound therapy (NPWT) in the field of wounds in the Web of Science core collection, and to explore the current status, frontiers, and trends of related research.Methods:The Web of Science core collection was used as the data source, and the search time period was from June 30, 2019 to June 30, 2024. Visual bibliometric analysis of the included literature was performed using VOSviewer 1.6.20 and CiteSpace 6.2.R4 for the number of publications, country and institution of publication, and keywords.Results:A total of 941 papers were included, with an increasing and then decreasing trend in the number of publications. The country with the largest number of publications was the United States (245), and although China ranked 2nd in the number of publications (208), the average number of citations in the literature was low. There was no strong cooperation network among publishing institutions. The proportion of articles published by core authors was 47.61% (448/941), with no significant core group of authors. In the past 5 years, research on NPWT in the field of wounds focused on chronic wound nursing and treatment, acute surgical incision complications and their prevention, wound closure and skin reconstruction, and risk factors for NPWT.Conclusions:The clinical application scope of NPWT has been expanded from simple wounds to complex wounds, tissue regeneration, and other fields. Researchers can carry out multi-center, large-sample, high-quality randomized controlled studies, enhance inter-institutional cooperation, and focus on interdisciplinary intersections and the formulation of NPWT care plans.
5.Host MYH9 protein promotes Zika virus invasion in U251 cells
Qianyi PENG ; Yalan FENG ; Jing HE ; Rong HUANG ; Jiafei ZHAN ; Yuhang ZHENG ; Chen CHEN ; Rong XIA ; Lei YUAN ; Jian YANG ; Kui XU
Chinese Journal of Experimental and Clinical Virology 2025;39(3):278-286
Objective:This study aimed to investigate the regulatory role and mechanism of myosin heavy chain 9 (MYH9) in the invasion of Zika virus (ZIKV) into human glioma cells (U251).Methods:Utilizing CRISPR/Cas9 technology, MYH9-knockout U251 cells (U251-MYH9 KD) were constructed. Following ZIKV infection, the protein expression levels, RNA load, and viral titer of ZIKV were detected through western blot (WB), Real-time fluorescence quantitative polymerase chain reaction (qPCR), and plaque formation assays, respectively. The infection efficiency of ZIKV in U251 cells treated with the MYH9 inhibitor blebbistatin was assessed. The binding and internalization efficiency of ZIKV were measured in U251-MYH9 KD cells. The interaction between MYH9 and the ZIKV envelope protein (E) was studied using co-immunoprecipitation (Co-IP). The effects of soluble MYH9 recombinant protein and anti-human MYH9 antibodies on ZIKV infection were evaluated by qPCR and plaque formation assays. Results:It was found that knockout or inhibition of MYH9 significantly suppressed ZIKV infection in U251 cells. MYH9 knockout notably inhibited the binding and internalization of ZIKV in U251 cells. MYH9 interacted with the ZIKV E protein, and both MYH9 recombinant protein and anti-human MYH9 antibodies, by blocking the binding of ZIKV E protein to cell surface MYH9, inhibited ZIKV infection in U251 cells in a dose-dependent manner.Conclusions:MYH9 facilitates ZIKV invasion into U251 cells through interaction with the ZIKV E protein.
6.Visual analysis of research hotspots and trends in negative-pressure wound therapy in the field of wounds in the past 5 years
Shiqi WEN ; Liping HUANG ; Jiafei LI ; Yuwen CHEN ; Ka LI
Chinese Journal of Modern Nursing 2025;31(26):3549-3555
Objective:To visualize and analyze the literature on the application of negative-pressure wound therapy (NPWT) in the field of wounds in the Web of Science core collection, and to explore the current status, frontiers, and trends of related research.Methods:The Web of Science core collection was used as the data source, and the search time period was from June 30, 2019 to June 30, 2024. Visual bibliometric analysis of the included literature was performed using VOSviewer 1.6.20 and CiteSpace 6.2.R4 for the number of publications, country and institution of publication, and keywords.Results:A total of 941 papers were included, with an increasing and then decreasing trend in the number of publications. The country with the largest number of publications was the United States (245), and although China ranked 2nd in the number of publications (208), the average number of citations in the literature was low. There was no strong cooperation network among publishing institutions. The proportion of articles published by core authors was 47.61% (448/941), with no significant core group of authors. In the past 5 years, research on NPWT in the field of wounds focused on chronic wound nursing and treatment, acute surgical incision complications and their prevention, wound closure and skin reconstruction, and risk factors for NPWT.Conclusions:The clinical application scope of NPWT has been expanded from simple wounds to complex wounds, tissue regeneration, and other fields. Researchers can carry out multi-center, large-sample, high-quality randomized controlled studies, enhance inter-institutional cooperation, and focus on interdisciplinary intersections and the formulation of NPWT care plans.
7.Analysis on pre-donation blood testing in blood services in Chongqing
Li MO ; Huayou DAI ; Jiafei YI ; Zhen ZHANG ; Wei DONG ; Xiaohong RAN ; Qingyu XIA ; Xia HUANG ; Tao HE
Chinese Journal of Blood Transfusion 2021;34(12):1279-1281
【Objective】 To provide data reference for the implementation of the homogenization of pre-donation blood testing by investigating the relevant situation of pre-donation blood testing in various blood services in Chongqing and analyzing their differences. 【Methods】 A questionnaire covering the basic information of pre-donation blood testing items, quality control and the management of deferral donors was developed, and issued to 19 blood services in Chongqing through E-mails by Chongqing Society of Blood Transfusion. The data collected were sorted, revised and analyzed. 【Results】 A total of 19 questionnaires from 19 blood services(including 1 blood center, 1 sub-center, 6 central blood stations and 11 central blood banks) were collected. All of the pre-donation blood test items of 19 blood services met the Blood Donor Health Test Requirements. Hemoglobin, blood group, ALT and HBsAg testing were carried out by 19 blood services, anti-TP testing by 15, and lipid blood testing by 11, using different detection methods and reagents. Significant differences were found in the frequency and rules of internal quality control for quantitative testing items. In addition, the deferral time and re-recruitment strategy of deferral blood donors were also significantly different. 【Conclusion】 There were differences in the management of pre-donation blood testing and blood donor management after blood donation among blood services in Chongqing. Further standardization was needed to realize regional homogenization and guarantee blood safety and the safety of blood donors.
8. Laparoscopic pancreaticoduodenectomy: a report of 233 cases by a single team
Weiwei JIN ; Xiaowu XU ; Yiping MOU ; Yucheng ZHOU ; Renchao ZHANG ; Jiafei YAN ; Jiayu ZHOU ; Chaojie HUANG ; Chao LU
Chinese Journal of Surgery 2017;55(5):354-358
Objective:
To summary the experience of 233 cases of laparoscopic pancreaticoduodenectomy (LPD) performed by a single surgical team.
Methods:
Data of patients undergoing LPD from September 2012 to October 2016 were reviewed. There were 145 males and 88 females with the mean age of(60.3±13.0)years old, ranging from 19 to 92 years old, and the mean body mass index of (22.8±3.5)kg/m2, ranging from 16.3 to 36.8 kg/m2. There were 195 patients with clinical manifestation and 54 patients who had the history of abdominal surgery.
Results:
LPD were performed on 233 patients by same surgical team consecutively. The mean operative time was(368.0±57.4)minutes. Mean blood loss was(203.8±138.6)ml. The postoperative morbidity rate was 33.5%, with 6.9% of grade B or C pancreatic fistula and 9.9% of bleeding. The reoperation rate was 5.6%. The mortality during 30 days after operation was 0.9%. Mean postoperative hospital stay was (18.1±11.2)days. Mean tumor size was (3.9±2.4)cm, and the mean number of lymph nodes harvested was 21.3±11.9.One hundred and sixty-three patients were diagnosed as malignant tumor, including pancreatic adenocarcinoma(
9. Effect of HIF-2α on regulating CDCP1 to promote hepatocellular carcinoma metastasis
Manqing CAO ; Junrong GAO ; Jiafei HUANG ; Abin YOU ; Zhigui GUO ; Hongyuan ZHOU ; Feng FANG ; Wei ZHANG ; Xiaolin ZHU ; Ti ZHANG
Chinese Journal of Oncology 2017;39(1):18-23
Objective:
To investigate the effect of hypoxia inducible factor 2α (HIF-2α) on regulating CUB domain-containing protein 1 (CDCP1) and its role in hepatocellular carcinoma metastasis.
Methods:
HIF-2α-knocked down and HIF-2α-stably overexpressing cells (MHCC97H) were prepared by small interfering RNA (siRNA) and lentivirus transfection, respectively. The expression of CDCP1 protein and mRNA in the above cells was detected by western blot and real-time PCR. The effect of HIF-2α on cell invasion ability was determined by Transwell assay. Furthermore, immunohistochemical staining was performed to detect the expression of CDCP1 in human HCC tissue samples.
Results:
Both HIF-2α and CDCP1 were induced under hypoxic conditions. The activation of CDCP1 under hypoxic conditions was dependent on the expression of HIF-2α.When HIF-2α was overexpressed, the mRNA level of CDCP1 was greatly upregulated (5.92±0.28,
10.Effect of different pancreaticojejunal anastomoses on the occur-rence of pancreatic fistula after pancreaticoduodenectomy
Jiafei HUANG ; Ti ZHANG ; Hongyuan ZHOU ; Huikai LI ; Yunlong CUI ; Qiang LI
Chinese Journal of Clinical Oncology 2015;42(8):441-445
Objective:To investigate the effect of pancreaticojejunostomy on the occurrence of postoperative pancreatic fistula (POPF) after pancreaticoduodenectomy. Methods:Data from 145 patients with periampullary tumor who underwent pancreaticoduode-nectomy in Tianjin Medical University Cancer Institute and Hospital between October 2008 and August 2013 were reviewed. Factors potentially associated with POPF were analyzed by Pearson chi-square test and Logistic regression analysis. Results:Among the 145 patients, 27 were diagnosed with POPF, including 5 grade A, 17 grade B and 5 grade C. Neither duct to mucosa nor Blumgart pancreati-cojejunostomy was correlated with POPF in grade C. The univariate analysis showed that gender, pancreatic cancer, portal vein involve-ment, type of pancreaticojejunostomy, texture of pancreas, and diameter of the main pancreatic duct were closely correlated with POPF. The multivariate analysis using Logistic regression showed that different pancreaticojejunal anastomoses and genders were independent predictors of POPF. Conclusion:Different types of pancreatic anastomoses are a risk factor for POPF after pancreaticoduodenectomy.

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