1.Mechanism by which exogenous basic fibroblast growth factor promotes wound healing in rats
Zhenchao LI ; Xiling DU ; Zhixin HAN ; Dawei NIU ; Changwei FAN
Chinese Journal of Tissue Engineering Research 2025;29(11):2243-2251
BACKGROUND:This study provided insight into the molecular mechanisms by which exogenous basic fibroblast growth factor(bFGF)promotes wound healing. OBJECTIVE:To investigate the effect of exogenous bFGF on macrophage phenotype transition and granulation regeneration during wound repair in rats. METHODS:(1)In vitro experiment:Cells were divided into normal control group,low-dose bFGF group,high-dose bFGF group,and bFGF+valproic acid group.100 and 200 μg/L bFGF was added into the cell culture medium of low-dose bFGF group and high-dose bFGF group,respectively,while 200 μg/L bFGF and 20 mmol/L valproic acid were added into the cell culture medium of valproic acid group.EdU test,scratch test and tubule formation test were used to detect the effects of bFGF on proliferation,migration and angiogenesis of human umbilical vein endothelial cells.(2)In vivo experiment:Sprague-Dawley rats were randomly divided into model group,low-dose bFGF group,high-dose bFGF group and bFGF+valproic acid group.The open wound model of full-thickness skin defect was established in low-dose bFGF group,high-dose bFGF group and bFGF+valproic acid group.Rats in the low-and high-dose bFGF groups were given 100 and 200 μg/L bFGF through subcutaneous injection,while those in the bFGF+valproic acid group received subcutaneous injection of 200 μg/L bFGF and intraperitoneal injection of 10 mg/kg valproic acid.The wound healing rate of rats was detected at 7 and 14 days of administration.TUNEL was used to detect the apoptosis of cells in wound tissue.Enzyme linked immunosorbent assay was used to detect the serum levels of malondialdehyde,superoxide dismutase,tumor necrosis factor-α and interleukin-10.Immunofluorescence detection was conducted to detect the phenotypic transformation of macrophages in wound tissue.Immunohistochemistry was used to detect the expression of proliferating cell nuclear antigen,platelet endothelial cell adhesion molecule-1(CD31)and vascular endothelial growth factor in wound tissue.Western blot was used to detect the expression of Notch1 and Jagged1 in wound tissue. RESULTS AND CONCLUSION:(1)Compared with the normal control group,bFGF could significantly promote the proliferation,migration and angiogenesis of human umbilical vein endothelial cells in a dose-dependent manner.(2)Compared with the model group,bFGF could significantly promote wound healing,downregulate the rate of apoptosis in wound tissue,decrease the levels of malondialdehyde and tumor necrosis factor-α in serum,increase the levels of superoxide dismutase and interleukin-10,promote the conversion of macrophages to type M2 in wound tissue,upregulate the expression of proliferating cell nuclear antigen,CD31 and vascular endothelial growth factor in wound tissue,and inhibit the expression of Notch1 and Jagged1 in a dose-dependent manner.Valproic acid could partially reverse the promoting effect of bFGF on wound healing.To conclude,bFGF can significantly promote wound healing and granulation regeneration and induce the conversion of macrophages to M2,which may be related to the regulation of Notch1/Jagged1 signaling pathway.
2.Evaluation of chemiluminescence immunoassay kit for detection of hepatitis D virus IgG antibody
Rongchen YUAN ; Fangming CHENG ; Kuanhui XIANG ; Yongcong LI ; Tianxun HUANG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Zhuanguo WANG ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Chungen QIAN ; Tong LI ; Tao SHEN ; Bangning CHENG
Chinese Journal of Laboratory Medicine 2024;47(3):234-238
Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.
3.Assessment and preliminary clinical application of a domestic nucleic acid detection reagent for hepatitis D virus
Yongcong LI ; Rongchen YUAN ; Kuanhui XIANG ; Guomin OU ; Tianxun HUANG ; Fangming CHENG ; Zhenchao TIAN ; Xiongwei LIU ; Xiaozhong WANG ; Feng GUO ; Yahong MA ; Jing ZHOU ; Erhei DAI ; Bangning CHENG ; Tong LI ; Tao SHEN ; Chungen QIAN
Chinese Journal of Laboratory Medicine 2024;47(3):239-244
Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.
4.Application of artificial intelligence based on multimodal fundus image data in the diagnosis and treatment of cardiovascular diseases
Yan WANG ; Xue HE ; Hanpeng ZHAO ; Cong LI ; Yun REN ; Jianrong JIANG ; Zhenchao DU ; Xiaohong YANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(09):1344-1350
Cardiovascular diseases is the leading cause of threat to human life and health worldwide. Early risk assessment, timely diagnosis, and prognosis evaluation are critical to the treatment of cardiovascular diseases. Currently, the evaluation of diagnosis and prognosis of cardiovascular diseases mainly relies on imaging examinations such as coronary CT and coronary angiography, which are expensive, time-consuming, partly invasive, and require high professional competence of the operator, making it difficult to promote in the community or in areas where medical resources are scarce. The fundus microcirculation is a part of the human microcirculation and has similar embryological origins and physiopathological features to cardiovascular circulation. Several studies have revealed fundus imaging biomarkers associated with cardiovascular diseases, and developed and validated intelligent diagnosis and treatment models for cardiovascular diseases based on fundus imaging data. Fundus imaging is expected to be an important adjunct to cardiovascular disease diagnosis and treatment given its noninvasive and convenient nature. The purpose of this review is to summarize the current research status, challenges, and future prospects of the application of artificial intelligence based on multimodal fundus imaging data in cardiovascular disease diagnosis and treatment.
5.A zero-sum game or an interactive frame? Iron competition between bacteria and humans in infection war.
Zhenchao WU ; Jiqi SHAO ; Jiajia ZHENG ; Beibei LIU ; Zhiyuan LI ; Ning SHEN
Chinese Medical Journal 2022;135(16):1917-1926
Iron is an essential trace element for both humans and bacteria. It plays a vital role in life, such as in redox reactions and electron transport. Strict regulatory mechanisms are necessary to maintain iron homeostasis because both excess and insufficient iron are harmful to life. Competition for iron is a war between humans and bacteria. To grow, reproduce, colonize, and successfully cause infection, pathogens have evolved various mechanisms for iron uptake from humans, principally Fe 3+ -siderophore and Fe 2+ -heme transport systems. Humans have many innate immune mechanisms that regulate the distribution of iron and inhibit bacterial iron uptake to help resist bacterial invasion and colonization. Meanwhile, researchers have invented detection test strips and coupled antibiotics with siderophores to create tools that take advantage of this battle for iron, to help eliminate pathogens. In this review, we summarize bacterial and human iron metabolism, competition for iron between humans and bacteria, siderophore sensors, antibiotics coupled with siderophores, and related phenomena. We also discuss how competition for iron can be used for diagnosis and treatment of infection in the future.
Humans
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Siderophores/metabolism*
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Iron/metabolism*
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Bacteria
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Anti-Bacterial Agents/pharmacology*
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Biological Transport
6.Effect of upregulated HuR gene on radiosensitivity of esophageal squamous cell carcinoma cell Kyse450
Dan HAN ; Lu LI ; Zhiwen KAN ; Zhenchao TAO ; Liting QIAN
Chinese Journal of Radiation Oncology 2022;31(5):456-461
Objective:To evaluate the effect of upregulation of HuR gene on the radiosensitivity of esophageal squamous cell carcinoma cell Kyse450. Methods:The HuR gene of Kyse450 cells was upregulated by lentivirus. At the same time, X-ray irradiation at a dose of 6 Gy was selected as the intervention condition. Western blot and qPCR were used to detect the expression levels of protein and RNA after Kyse450 transfection, respectively. CCK8 kit was employed to determine the cell proliferation rate. Clone formation assay was adopted to evaluate the ability of cell clone formation. Wound healing experiment and the Transwell test were performed to detect changes in cell migration. Results:CCK8 assay showed that the proliferation ability of cells was enhanced after upregulation of HuR gene, and this enhancement trend was more obvious after radiation. The plate cloning experiment showed that with the increase of radiation dose, the clone formation rates were decreased in both groups, but the clone formation rate in the overexpression group was higher than that in the control group. Wound healing experiment and Transwell test demonstrated that the wound healing rate and migration ability in the overexpression group were higher than those in the control group, and the difference was more significant after radiotherapy. Western blot showed that the levels of MMP9 and MMP2 at 24 h after radiotherapy in the overexpression group were higher than those in the control group. Conclusion:The upregulation of HuR can enhance the proliferation, cloning, migration capabilities and decrease the radiosensitivity of esophageal squamous cell carcinoma cells.
7.Value of MRI in the differential diagnosis of uveal melanoma and other intraocular masses in adults
Mingxia SUN ; Qinghua CHEN ; Xiaolin XU ; Zhenchao SUN ; Yaping SU ; Bin LI ; Wenbin WEI ; Junfang XIAN
Chinese Journal of Radiology 2020;54(3):181-186
Objective:To compare MRI findings of uveal melanomas and other intraocular masses in adults and to evaluate the optimal MRI features for the differential diagnosis.Methods:The MRI data of 254 patients with intraocular masses confirmed by histopathologic or clinical follow-up results from March 2005 to December 2015 in Beijing Tongren Hospital, Capital Medical University was analyzed retrospectively. A total of 128 patients with uveal melanoma and 126 patients with other intraocular masses were enrolled. MRI findings of the masses including the location, shape, margin, signal intensity on T 1WI and T 2WI compared to the gray matter and vitreous body, homogeneity and degree of enhancement of the mass, secondary retinal detachment, height, and ratio of basal diameter to height were compared between uveal melanoma and other intraocular masses by χ 2 test or independent t test. The logistic regression analysis was performed to identify the most discriminating MRI features. The diagnostic performance of different predictive models was analyzed by receiver operating characteristic (ROC) curve. The model of multiple parameters was established by logistic analysis, and the diagnostic efficacy was evaluated. Results:The location, shape, margin, signal intensity on T 1WI and T 2WI compared to the gray matter and vitreous body, homogeneity and degree of enhancement of the mass, secondary retinal detachment, height and ratio of basal diameter to height were significantly different between the uveal melanoma and other intraocular masses (all P<0.05). The results of logistic regression analysis showed that the signal intensity on T 1WI and T 2WI compared to the gray matter, height, shape, location, degree of enhancement of the mass, and secondary retinal detachment were associated with higher likelihood of uveal melanoma. The mass with hypointense on T 2WI (compared with gray matter) was the best MRI feature in the differential diagnosis of uveal melanoma and other intraocular masses (odds ratio 12.237), with the accuracy of 86.2%. The diagnostic accuracy of low signal on T 2WI and high signal on T 1WI (compared with gray matter) was significantly higher than those of vitreous (Delong test, P<0.001). The sensitivity, specificity and accuracy of the combination of height, degree of enhancement, T 1WI and T 2WI signal compared to the gray matter were 90.6%, 92.1% and 96.1%, respectively. The diagnostic performance of combination features was higher than that of each single sign ( P<0.001). Conclusion:MRI features including the signal intensity on T 1WI and T 2WI compared to the gray matter, height, shape, degree of enhancement of the mass, and secondary retinal detachment are helpful in the differential diagnosis between uveal melanoma and other intraocular masses. In addition, the gray matter as the reference is superior to the vitreous body in the evaluation of the signal intensity of intraocular masses.
8.Monte Carlo-based simulation of influence of linear accelerator beam parameter on percentage depth dose
Fan ZHANG ; Haibiao WU ; Ainong XIAO ; Xiaohong AI ; Manbo CAI ; Pian LI ; Lixiang CHENG ; Zhenchao ZHANG ; Guopu QU
Chinese Journal of Radiological Medicine and Protection 2018;38(2):145-149
Objective To determine the optimal electron beam energy at different field size through a Monte Carlo-based simulation of the therapy head of Varian X 6 MV linear accelerator so as to study the influence of radial intensity on depth dose.Methods Firstly,keeping the radial intensity unchanged for the field of interest while changing electron beam energy,compassion was carried out of calculated percentage depth doses between measured values.Thus,the optimal energy was identified for this field size.Then,the obtained energy was set the optimal value to study the radial intensity influence on the depth doses.Results The optimal electron energy for 4 cm ×4 cm,10 cm × 10 cm,20 cm × 20 cm and 30 cm × 30 cm field sizes was 5.9,6.0,6.3 and 6.4 MeV respectively.Changes in radial intensities resulted in negligible changes in percentage depth doses for4 cm ×4-cm and 10 cm × 10 cm fields,but led to observable discrepancy for 20 cm × 20 cm and 30 cm × 30 cm fields.Conclusions The optimal electron energies for different field sizes are slightly different.Change in radial intensity distribution has significant influence on the depth dose for large field.To improve simulation accuracy,the field size needs to be taken into consideration in determining the electron beam energy and radial intensity distribution.
9.Changes of mouse hippocampal neurogenesis during aging
Pinyue LIU ; Zhenchao JIN ; Xiaolan DENG ; Li YUAN ; Min ZHENG ; Yaping WANG ; Jian WANG ; Rong JIANG
Chinese Journal of Tissue Engineering Research 2016;20(11):1564-1569
BACKGROUND:Abnormal hippocampal neurogenesis during aging has been reported to result in learning and memory dysfunction. But its mechanism is unclear.
OBJECTIVE: To understand the changes of mouse neurogenesis in the hippocampal subgranular zone during aging.
METHODS:C57BL/6 mice 2, 6 and 20 months of age were used. Immunochemistry was used to count the number of neural stem cels (nestin+), neuroblasts (Doublecortin+, DCX+), and proliferative cels (proliferating cel nuclear antigen+, PCNA+) in the hippocampal subgranular zone. mRNA expressions of aging-related genes, p19Arf and p21Cip1/Waf1, in the hippocampus were detected by reverse transcription-PCR.
RESULTS AND CONCLUSION: Compared with the young and middle age groups, the number of PCNA+ cels, nestin+ and DCX+ cels in the hippocampal subgranular zone of the aged group decreased dramaticaly; the expression of p19Arf and p21Cip1/Waf1 mRNA increased in the aged group. Proliferation activity, the number of neural stem cels and neuronal differentiation al decreased. These findings indicate that the decline of hippocampal neurogenesis may be associated with increased expression of aging-related genes p19Arf and p21Cip1/Waf1 in the p19Arf-Mdm2-p53-p21Cip1/Waf1pathway.
10.Acute and 28-day sub-acute oral toxicity evaluation of two dietary bamboo charcoal powders in Sprague-Dawley rats.
Zhen-chao JIA ; Sha LUO ; Yu-ting ZHONG ; Xiao LI ; Jin-yao CHEN ; Li-shi ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(2):192-199
No data were available on the acute oral toxicity, short-term oral toxicity of vegetable carbon in animals. This study was designed to evaluate the safety of two commercially available dietary bamboo charcoal powders (BCP1 and BCP2). The size distribution of the two powders was determined by a Mastersizer 2000 laser particle size analyzer prior to the in vivo safety studies. For the acute toxicity study, a single dose of 11.24 g/kg body weight of BCP1 and BCP2 was given once orally to healthy Sprague-Dawley (SD) rats. Mortality and clinical symptoms were observed and recorded for the first 30 min after treatment, at 4 h post-administration, and then at least once daily for 14 days after administration. In the repeated dose 28-day oral toxicity study, BCP1 and BCP2 were administered orally at doses of 2.81, 5.62, and 11.24 g/kg body weight for 28 days to SD rats. Animals were sacrificed and organs and blood samples were analyzed. Results showed that both BCP1 and BCP2 were micro-sized and various in size. In the acute toxicity and the repeated dose 28-day oral toxicity studies, BCP caused neither mortality nor visible signs of toxicity in rats. No significant differences were found in the relative organ weights or in biochemical parameters in BCP treated groups compared to a control group. No treatment-related histological changes were observed in the organs of these animals. Based on these data, it is concluded that the median lethal dose (LD50) of BCP for both male and female rats is more than 11.24 g/kg body weight and the no-observed-adverse-effect level (NOAEL) is >11.24 g/kg body weight for 28 days.
Animals
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Bambusa
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chemistry
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Diet
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Female
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Male
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Powders
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Rats
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Rats, Sprague-Dawley
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Toxicity Tests, Acute

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