2.Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells
Chang LIU ; Jing YU ; Yongjun DU ; Yu XIE ; Xiaofei SONG ; Chang LIU ; Yan YAN ; Yue WANG ; Junfang QIN
Cancer Research and Treatment 2025;57(2):457-477
Purpose:
Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear.
Materials and Methods:
The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2.
Results:
LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects.
Conclusion
This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.
3.Long-term protective efficacy of recombinant CHO cell-derived hepatitis B vaccine:A 26-year follow-up study in Zhengding county
Qiudong SU ; Zhiyong HAO ; Feng QIU ; Lili QIAO ; Huijuan JIA ; Shuang ZHANG ; Pengpeng YUAN ; Juan CHU ; Zhiyong ZHANG ; Junfang YAO ; Jinling WEI ; Yan WANG ; Peiqi ZHAO ; Yujiao LIU ; Zhao GAO ; Jingchen MA ; Feng WANG ; Haisong ZHOU
Chinese Journal of Experimental and Clinical Virology 2025;39(4):434-440
Objective:To evaluate the long-term protective efficacy of the recombinant Chinese hamster ovary cell-derived hepatitis B vaccine(CHO-HepB)26 years post-vaccination in the rural China.Methods:Zhengding county,Hebei province was designated as a rural monitoring site for CHO-HepB efficacy. Study participants included individuals born between 1997 and 1999 who had completed the three-dose CHO-HepB primary series without booster doses. A cross-sectional survey was conducted in late 2024 using random sampling. Demographic and vaccination history data were collected via questionnaires,and hepatitis B virus(HBV)serological markers were detected using chemiluminescence. Historical surveillance data were integrated to infer infection statuses of HBsAg-positive individuals and evaluate longitudinal trends in anti-HBs seropositivity and antibody titers.Results:Among 178 participants(mean time since vaccination:26.2 years),the seroprevalence rates were 0.6% for HBsAg(95% CI:0.0%-1.6%),64.6% for anti-HBs(95% CI:57.6%-71.6%),and 1.1% for anti-HBc(95% CI:0.0%-2.7%). Compared to the pre-vaccination baseline HBsAg positivity of 11.3% in children under 10 years of age,the estimated vaccine protection rate was 95%. Two notable cases were identified:one with concurrent HBsAg and anti-HBc positivity and one with anti-HBs and anti-HBc positivity,suggestive of transient HBV exposure(1999—2009)without chronicity. Natural immune boosting was inferred for the latter case based on anti-HBs titer dynamics. Longitudinal analysis of four prior cross-sectional surveys(2005,2009,2013,and 2017)revealed no significant upward trends in HBsAg and anti-HBc positivity(both P>0.05)over 26 years,while anti-HBs seropositivity declined significantly( P<0.05)from 6 to 26 years post-vaccination. Conclusion:The CHO-HepB vaccine demonstrates sustained immunological persistence and robust long-term protection up to 26 years post-immunization. Continued emphasis on rigorous implementation of mother-to-child transmission prevention strategies is critical for future hepatitis B control.
4.Clinical value of detecting levels of plasma ceramide in familial homozygous hypercholesterolemia patients by liquid chromatography-tandem mass spectrometry
Yuxuan ZHANG ; Xinye YAO ; Luya WANG ; Yan CHEN ; Yan WANG ; Junfang WU ; Jun ZHANG
Chinese Journal of Laboratory Medicine 2025;48(7):879-887
Objective:To quantitatively detect plasma ceramide (Cer) levels using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyze plasma ceramide levels in homozygous familial hypercholesterolemia (HoFH) patients.Methods:A case-control study was conducted. Plasma samples were collected from 70 healthy individuals who underwent physical examinations at the Health Check-Up Center of Suzhou Municipal Hospital between October 2023 and December 2023, as well as from 34 HoFH patients (including 11 patients with concomitant coronary artery disease) who were treated in the Cardiology Department of the same hospital during the same period. Deuterated isotopes were used as internal standards, and a Phenomenex Kinetex C18 column (2.1×100 mm, 2.6 μm) was employed for separation. An LC-MS/MS method was developed using an AB Sciex 6500+triple quadrupole mass spectrometer for the detection of ceramides, followed by method validation. Additionally, the differences in ceramide levels among healthy individuals, HoFH patients, and HoFH patients with concomitant coronary artery disease were compared. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic value of various ceramides for HoFH.Results:A method for the quantification of 9 plasma ceramides was established, showing excellent linearity ( r>0.99), precision (1.8%-11.0%), and recovery rates (89.9%-114.7%), meeting the requirements for detection. Compared to the control group, the HoFH group exhibited significantly higher levels of Cer(d18∶1/14∶0), Cer(d18∶1/16∶0), Cer (d18∶1/18∶0), Cer (d18∶1/18∶1), Cer (d18∶1/20∶0), Cer(d18∶1/22∶0), Cer(d18∶1/24∶0), and Cer(d18∶1/24∶1) ( P<0.05). ROC curve analysis revealed that Cer(d18∶1/18∶0) had the highest diagnostic value for HoFH, with an area under the curve (AUC) of 0.83, a cutoff value of 64.6 ng/ml, and sensitivity and specificity of 82.4% and 87.1%, respectively. In HoFH patients complicated with coronary artery disease, the plasma ceramide levels of Cer(d18∶1/18∶0) [(155.7±25.1) ng/ml vs (103.0±11.1) ng/ml, P=0.03] and the ratio of Cer (d18∶1/18∶0) to Cer(d18∶1/24∶0) (0.077±0.012 vs 0.048±0.006, P=0.03) were significantly higher than in HoFH patients without coronary artery disease. Conclusion:A quantitative LC-MS/MS method was developed for detecting ceramides in human plasma, which shows potential diagnostic value for patients with HoFH.
5.Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells
Chang LIU ; Jing YU ; Yongjun DU ; Yu XIE ; Xiaofei SONG ; Chang LIU ; Yan YAN ; Yue WANG ; Junfang QIN
Cancer Research and Treatment 2025;57(2):457-477
Purpose:
Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear.
Materials and Methods:
The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2.
Results:
LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects.
Conclusion
This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.
6.Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells
Chang LIU ; Jing YU ; Yongjun DU ; Yu XIE ; Xiaofei SONG ; Chang LIU ; Yan YAN ; Yue WANG ; Junfang QIN
Cancer Research and Treatment 2025;57(2):457-477
Purpose:
Leptin interacts not only with leptin receptor (LEPR) but also engages with other receptors. While the pro-oncogenic effects of the adrenergic receptor β2 (ADRB2) are well-established, the role of leptin in activating ADRB2 in triple-negative breast cancer (TNBC) remains unclear.
Materials and Methods:
The pro-carcinogenic effects of LEPR were investigated using murine TNBC cell lines, 4T1 and EMT6, and a tumor-bearing mouse model. Expression levels of LEPR, NADPH oxidase 4 (NOX4), and ADRB2 in TNBC cells and tumor tissues were analyzed via western blot and quantitative real-time polymerase chain reaction. Changes in reactive oxygen species (ROS) levels were assessed using flow cytometry and MitoSox staining, while immunofluorescence double-staining confirmed the co-localization of LEPR and ADRB2.
Results:
LEPR activation promoted NOX4-derived ROS and mitochondrial ROS production, facilitating TNBC cell proliferation and migration, effects which were mitigated by the LEPR inhibitor Allo-aca. Co-expression of LEPR and ADRB2 was observed on cell membranes, and bioinformatics data revealed a positive correlation between the two receptors. Leptin activated both LEPR and ADRB2, enhancing intracellular ROS generation and promoting tumor progression, which was effectively countered by a specific ADRB2 inhibitor ICI118551. In vivo, leptin injection accelerated tumor growth and lung metastases without affecting appetite, while treatments with Allo-aca or ICI118551 mitigated these effects.
Conclusion
This study demonstrates that leptin stimulates the growth and metastasis of TNBC through the activation of both LEPR and ADRB2, resulting in increased ROS production. These findings highlight LEPR and ADRB2 as potential biomarkers and therapeutic targets in TNBC.
7.Long-term protective efficacy of recombinant CHO cell-derived hepatitis B vaccine:A 26-year follow-up study in Zhengding county
Qiudong SU ; Zhiyong HAO ; Feng QIU ; Lili QIAO ; Huijuan JIA ; Shuang ZHANG ; Pengpeng YUAN ; Juan CHU ; Zhiyong ZHANG ; Junfang YAO ; Jinling WEI ; Yan WANG ; Peiqi ZHAO ; Yujiao LIU ; Zhao GAO ; Jingchen MA ; Feng WANG ; Haisong ZHOU
Chinese Journal of Experimental and Clinical Virology 2025;39(4):434-440
Objective:To evaluate the long-term protective efficacy of the recombinant Chinese hamster ovary cell-derived hepatitis B vaccine(CHO-HepB)26 years post-vaccination in the rural China.Methods:Zhengding county,Hebei province was designated as a rural monitoring site for CHO-HepB efficacy. Study participants included individuals born between 1997 and 1999 who had completed the three-dose CHO-HepB primary series without booster doses. A cross-sectional survey was conducted in late 2024 using random sampling. Demographic and vaccination history data were collected via questionnaires,and hepatitis B virus(HBV)serological markers were detected using chemiluminescence. Historical surveillance data were integrated to infer infection statuses of HBsAg-positive individuals and evaluate longitudinal trends in anti-HBs seropositivity and antibody titers.Results:Among 178 participants(mean time since vaccination:26.2 years),the seroprevalence rates were 0.6% for HBsAg(95% CI:0.0%-1.6%),64.6% for anti-HBs(95% CI:57.6%-71.6%),and 1.1% for anti-HBc(95% CI:0.0%-2.7%). Compared to the pre-vaccination baseline HBsAg positivity of 11.3% in children under 10 years of age,the estimated vaccine protection rate was 95%. Two notable cases were identified:one with concurrent HBsAg and anti-HBc positivity and one with anti-HBs and anti-HBc positivity,suggestive of transient HBV exposure(1999—2009)without chronicity. Natural immune boosting was inferred for the latter case based on anti-HBs titer dynamics. Longitudinal analysis of four prior cross-sectional surveys(2005,2009,2013,and 2017)revealed no significant upward trends in HBsAg and anti-HBc positivity(both P>0.05)over 26 years,while anti-HBs seropositivity declined significantly( P<0.05)from 6 to 26 years post-vaccination. Conclusion:The CHO-HepB vaccine demonstrates sustained immunological persistence and robust long-term protection up to 26 years post-immunization. Continued emphasis on rigorous implementation of mother-to-child transmission prevention strategies is critical for future hepatitis B control.
8.Clinical value of detecting levels of plasma ceramide in familial homozygous hypercholesterolemia patients by liquid chromatography-tandem mass spectrometry
Yuxuan ZHANG ; Xinye YAO ; Luya WANG ; Yan CHEN ; Yan WANG ; Junfang WU ; Jun ZHANG
Chinese Journal of Laboratory Medicine 2025;48(7):879-887
Objective:To quantitatively detect plasma ceramide (Cer) levels using high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyze plasma ceramide levels in homozygous familial hypercholesterolemia (HoFH) patients.Methods:A case-control study was conducted. Plasma samples were collected from 70 healthy individuals who underwent physical examinations at the Health Check-Up Center of Suzhou Municipal Hospital between October 2023 and December 2023, as well as from 34 HoFH patients (including 11 patients with concomitant coronary artery disease) who were treated in the Cardiology Department of the same hospital during the same period. Deuterated isotopes were used as internal standards, and a Phenomenex Kinetex C18 column (2.1×100 mm, 2.6 μm) was employed for separation. An LC-MS/MS method was developed using an AB Sciex 6500+triple quadrupole mass spectrometer for the detection of ceramides, followed by method validation. Additionally, the differences in ceramide levels among healthy individuals, HoFH patients, and HoFH patients with concomitant coronary artery disease were compared. Receiver operating characteristic (ROC) curves were used to evaluate the diagnostic value of various ceramides for HoFH.Results:A method for the quantification of 9 plasma ceramides was established, showing excellent linearity ( r>0.99), precision (1.8%-11.0%), and recovery rates (89.9%-114.7%), meeting the requirements for detection. Compared to the control group, the HoFH group exhibited significantly higher levels of Cer(d18∶1/14∶0), Cer(d18∶1/16∶0), Cer (d18∶1/18∶0), Cer (d18∶1/18∶1), Cer (d18∶1/20∶0), Cer(d18∶1/22∶0), Cer(d18∶1/24∶0), and Cer(d18∶1/24∶1) ( P<0.05). ROC curve analysis revealed that Cer(d18∶1/18∶0) had the highest diagnostic value for HoFH, with an area under the curve (AUC) of 0.83, a cutoff value of 64.6 ng/ml, and sensitivity and specificity of 82.4% and 87.1%, respectively. In HoFH patients complicated with coronary artery disease, the plasma ceramide levels of Cer(d18∶1/18∶0) [(155.7±25.1) ng/ml vs (103.0±11.1) ng/ml, P=0.03] and the ratio of Cer (d18∶1/18∶0) to Cer(d18∶1/24∶0) (0.077±0.012 vs 0.048±0.006, P=0.03) were significantly higher than in HoFH patients without coronary artery disease. Conclusion:A quantitative LC-MS/MS method was developed for detecting ceramides in human plasma, which shows potential diagnostic value for patients with HoFH.
9.Correlations of serum cell division cycle 42, β2-microglobulin and fibulin 1 levels with abdominal aortic calcification in patients with maintenance hemodialysis
Junfang YAN ; Qian ZONG ; Liang YUAN ; Ting BAO ; Wenting XU
Journal of Clinical Medicine in Practice 2024;28(18):47-50
Objective To investigate the correlations of serum cell division cycle 42 (CDC-42), β2-microglobulin (β2-MG) and fibulin 1 (FBLN1) levels with abdominal aortic calcification (AAC) in patients with maintenance hemodialysis (MHD). Methods General information and biochemical indicators of 74 MHD patients were collected, and they were divided into no to mild AAC group (
10.Vascular embolization induced by injection of autologous fat in rabbits
Yan OU ; Yingjie LUO ; Junfang LIANG ; Shufang LIANG ; Chenggang YI ; Sha LIU
Chinese Journal of Medical Aesthetics and Cosmetology 2024;30(3):286-291
Objective:To explore the severe complications, such as blindness, brain infarction and even death caused by cosmetic autologous fat filler injection and their underlying mechanisms.Methods:From May 2022 to October 2023, 64 male New Zealand rabbits were selected in the clinical laboratory of Shaanxi University of Chinese Medicine and divided into 8 groups with 8 rabbits in each group. They were divided into grinding fat group, fat granule group, fat lipid group and normal saline group, and each group was further divided into 0.2 ml group and 0.4 ml group. Fat was cut from the rabbit groin, then chopped or treated with collagenase I, and centrifuged to separate fat lipids and fat particles, as well as other tissues. The rabbit facial artery was exposed along the incision below the mandibular angle, and 0.2 or 0.4 ml of chopped fat, fat particles, and lipids were retrogradely injected into the facial artery in each group, and then the incision was closed under the microscope. Ophthalmic and neurological symptoms were observed and recorded after surgery, and visual electrophysiology and fundus microscopy were performed to verify visual acuity and fundus artery obstruction.Results:Two weeks after surgery, the incidence of ophthalmic symptoms in the 0.2 ml injection group was 100% in the grinding fat group, 62.5% in the fat granule group, 0 in the lipid group and 0 in the normal saline group. The complication rates of 0.4 ml embolic injection were 100%, 87.50%, 12.5% and 0, respectively. The incidence of neurological symptoms was 62.5%, 25.0%, 0 and 0, respectively. Mortality rates were 37.5%, 12.5%, 0 and 0 after injection of 0.2 ml, and 100%, 50%, 0 and 0 after injection of 0.4 ml, respectively.Conclusions:Animal models have shown that grinding adipose tissue without collagenase I treatment is more likely to cause serious complications, simple lipid entry into blood vessels does not block relevant arteries, and fat volume is positively correlated with the incidence of postoperative complications.


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