1.Whole-genome polymorphism of CD36 by third-generation sequencing technology
Jing LIU ; Xiuzhang XU ; Haoqiang DING ; Jing DENG ; Yangkai CHEN ; Wenjie XIA ; Xin YE
Chinese Journal of Blood Transfusion 2025;38(5):610-614
Objective: To analyze CD36 gene by PacBio Sequel Ⅱ the third-generation sequencing technology (TGS), including non-coding sequence, and to investigate the molecular mechanism of CD36 deficiency. Methods: Flow cytometry was performed in the southern Chinese population to detect the CD36 phenotype. Among them, 15 cases of CD36 type I deficiency, 15 cases of CD36 type Ⅱ deficiency, and 10 positive samples were selected. The TGS of the CD36 gene was performed and statistical analysis was conducted. Results: 40 samples (including 15 cases of type I deficiency, 15 cases of type Ⅱ deficiency, and 10 positive samples) were subjected by TGS of CD36 full-length sequences (except part of intron1). A total of 180 polymorphic loci were identified. Among them, 13 kinds were in the coding region, the rest were in non-coding region, with most mutations located in regulatory regions such as the 5′-UTR and 3′-UTR. Conclusion: The high polymorphism of CD36 non-coding regions, particularly in regulatory sequences, provides mechanistic insights into type Ⅱ CD36 deficiency.
2.In vitro expression analysis of the ITGB3 c.598G/A mutation and its association with FNAIT
Haoqiang DING ; Xin YE ; Xiuzhang XU ; Wenjie XIA ; Jing DENG ; Jing LIU ; Yangkai CHEN ; Dawei CHEN ; Yaori XU
Chinese Journal of Blood Transfusion 2025;38(7):873-878
Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B
57∶01 and A
02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.
3.In vitro expression analysis of the ITGB3 c.598G/A mutation and its association with FNAIT
Haoqiang DING ; Xin YE ; Xiuzhang XU ; Wenjie XIA ; Jing DENG ; Jing LIU ; Yangkai CHEN ; Dawei CHEN ; Yaori XU
Chinese Journal of Blood Transfusion 2025;38(7):873-878
Objective: To explore the role of the c.598G>A mutation of the ITGB3 gene in the occurrence of fetal and neonatal alloimmune thrombocytopenia (FNAIT) through its expression in vitro. Methods: The platelet antibodies in the sera of the affected neonate and her mother were detected using commercial enzyme-linked immunosorbent assay (ELISA), solid-phase agglutination, flow cytometry and the gold standard monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The common human platelet antigen (HPA) genotypes of the neonate and her parents were obtained using the HPA-SSP method. The presence of mutations was analyzed by sequencing the exons of the ITGB3 and ITGA2B genes. The target gene of ITGB3 was obtained by PCR amplification using the existing human platelet cDNA. The wild-type ITGB3 eukaryotic expression vector was constructed by TA cloning technology. The 598G>A mutant ITGB3 eukaryotic expression vector was obtained by point mutation, and the plasmid DNA was co-transfected with that of ITGA2B (αⅡb) into HEK293 cells. The transfected cells stably expressing GP Ⅱb/Ⅲa were screened and obtained. The expression of GP Ⅱb/Ⅲa in 598G>A mutant transfected cells and the presence of antibodies against this mutation in the serum of mother were detected by flow cytometry and MAIPA. Results: Antibodies against HLA-class Ⅰ and GP Ⅱb/Ⅲa glycoproteins were detected in the serum of the neonate's mother, and subsequent HLA antibody-specific testing confirmed the presence of antibodies against HLA-B
57∶01 and A
02∶05. ITGB3 sequencing showed that the neonate and her father carried the c.598G>A point mutation, which results in the change of glutamate to lysine at position 200. Antibodies against GP Ⅱb/Ⅲa glycoproteins were not detected using constructed c.598G>A mutant transfected cells reacted with the maternal serum. Conclusion: The in vitro expression and analysis of the ITGB3 c.598G>A mutation did not support a role for this mutation in the pathogenesis of FNAIT. The establishment of this method facilitates the discovery of new platelet low-frequency antigens, and provides a theoretical foundation for the detection of antibodies against platelet antigens associated with patients with adverse pregnancy and childbirth histories.
4.Predicting mortality risk in severe ards patients using indirect calorimetry-based oxygen consumption and carbon dioxide production rates
Ke GUAN ; Huihuang ZOU ; Yuna HU ; Ling YE ; Yanwei CHENG ; Jingjing NIU ; Cunzhen WANG ; Ke QIN ; Tingyuan ZHANG ; Bin YANG ; Yuhan SUN ; Wenliang ZHU ; Qingbo FAN ; Zhisong GUO ; Yongchun CHEN ; Wenjie WANG
Chinese Journal of Emergency Medicine 2025;34(3):396-403
Objective:To investigate the relationship between oxygen consumption (VO 2), carbon dioxide production (VCO 2), and Oxygen Consumption/lactate (VO 2/Lac) with risk of death in patients with severe ARDS. Methods:A retrospective cohort study method was used, and the study subjects were hospitalized for >5 days adult patients with severe ARDS in the central intensive care unit of Henan Provincial People's Hospital from 1 March 2020 to 30 June 2023. The following patients were excluded: IC test was not completed on the 4th day of ICU admission, IC test results were unreliable, mechanical ventilation duration had exceeded 48 h at the time of ICU transfer or admission, palliative care patients and pregnant and parturient women. Using indirect calorimetry to determine VO 2 and VCO 2 values on the 4th day of admission, reviewing medical records to obtain general condition, disease information, blood gas analysis (including lactate value), diagnostic and therapeutic measures, and following up deaths by telephone and time of death. The primary outcome measure was death at 90 days, and the secondary outcome measure was death at 28 days, length of stay in ICU, total length of stay, and total hospitalization cost. Cox regression analysis and linear regression analysis were used to investigate the relationship between VO 2, VCO 2, VO 2/Lac and primary and secondary outcome indexes. Results:A total of 216 patients were enrolled, 78 patients (36.1%) died and 138 patients (63.9%) survived at 90 days. After correction for confounders, the results of multifactorial Cox regression analysis suggested that compared with the Q4 group, HR (95% CI) for 90-day risk of death in the VO 2 Q1 and Q2 groups was 3.21 (1.38, 7.49) and 3.24 (1.42, 7.38), and HR (95% CI) for 90-day risk of death in the VCO 2 Q1, Q2 and Q3 groups was 5.88 (2.33, 14.84), 4.26 (1. 60, 11.34) and 3.54 (1.34, 9.35), respectively, and the HR (95% CI) for 90-day risk of death in the VO 2/Lac Q1, Q2 and Q3 groups were 8.72 (3.01, 25.25), 8.43 (2.91, 24.47) and 4.04 (1.34, 12.17) respectively. P-trends were all <0.05, indicating that VO 2, VCO 2 and VO 2/Lac were linearly and negatively associated with the risk of 90-day mortality. In addition, VO 2, VCO 2, and VO 2/Lac were negatively associated with 28-day risk of death and higher VO 2/Lac was negatively associated with length of ICU stay. Conclusions:VO 2, VCO 2 and VO 2/Lac were negatively associated with 90-day mortality risk and 28-day mortality risk in patients with severe ARDS and may be independent risk factors predicting mortality risk of such patients.
5.Brain-computer interface combined with different therapies for limb dysfunction in stroke patients:effectiveness and mechanism analysis
Xuesong WANG ; Yue WANG ; Yan XU ; Wenhui ZENG ; Wenming LU ; Xingkun TANG ; Wenjie CHEN ; Junsong YE
Chinese Journal of Tissue Engineering Research 2025;29(30):6538-6546
BACKGROUND:In recent years,brain-computer interface technology has shown significant promise for rehabilitating limb dysfunction in stroke patients.With ongoing research deepening and its broader clinical application,combining brain-computer interface with other rehabilitation therapies to improve limb function has become a focal point of study.OBJECTIVE:To analyze and summarize the efficacy of brain-computer interface combined with various therapies in treating limb dysfunction in stroke patients and to explore the clinical value of these combined strategies.METHODS:The search terms used for the literature review in Chinese databases were"brain-computer interface,BCI,stroke,"while the terms"brain-computer interface,BCI,brain-computer interaction,brain-machine interface,BMI,stroke"were used for English databases.Literature searches were conducted in CNKI,WanFang,VIP,PubMed,Embase,and Web of Science,from the time of database construction to September 2024.Finally,a total of 3 054 articles were retrieved,and 75 articles were included after screening for summarization.RESULTS AND CONCLUSION:Currently,brain-computer interfaces,used alone or in combination with other treatments such as Chinese medical treatment,conventional rehabilitation therapy,or physical factor therapy,are achieving better outcomes in treating limb dysfunction in stroke patients.However,the efficacy of brain-computer interfaces combined with transcranial direct current stimulation for treating upper and lower limb dysfunctions is still debated.Researchers are increasingly recognizing the feasibility of these combined therapies.Yet,challenges such as limited exploration of therapeutic mechanisms,absence of standardized protocols,and small sample sizes hinder their broad application.Future research should therefore focus on understanding the mechanisms by which brain-computer interfaces can enhance effects when combined with other therapies and on standardizing criteria for clinical trials to enable widespread clinical adoption.
6.Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury (version 2025)
Aijun XU ; Shuixia LI ; Bo CHEN ; Mengyuan YE ; Lejiao LANG ; Ning NING ; Lin ZHANG ; Changqing LIU ; Zhonglan CHEN ; Weihu MA ; Weishi LI ; Xiaoning WANG ; Dongmei BIAN ; Jiancheng ZENG ; Xin WANG ; Yuan GAO ; Yaping CHEN ; Jiali CHEN ; Yun HAN ; Xiuting LI ; Yang ZHOU ; Xiaojing SU ; Qiong ZHANG ; Tianwen HUANG ; Ping ZHANG ; Hua LIN ; Xingling XIAO ; Ruifeng XU ; Fanghui DONG ; Bing HAN ; Luo FAN ; Yanling PEI ; Suyun LI ; Xiaoju TAN ; Rongchen GUO ; Yefang ZOU ; Xiaoyun HAN ; Junqin DING ; Yi WANG ; Shuhua DENG ; Jinli GUO ; Yinhua LIANG ; Yuan CEN ; Xiaoqin LIU ; Junru CHEN ; Haiyang YU ; Lunlan LI ; Ying REN ; Yunxia LI ; Jianli LU ; Ying YING ; Lan WEI ; Yin WANG ; Qinhong XU ; Yanqin ZHANG ; Yang LYU ; Shijun ZHANG ; Sui WENJIE ; Sanlian HU ; Shuhong YANG ; Guoqing LI ; Jingjing AN ; Baorong HE ; Leling FENG
Chinese Journal of Trauma 2025;41(6):530-541
Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.
7.Brain-computer interface combined with different therapies for limb dysfunction in stroke patients:effectiveness and mechanism analysis
Xuesong WANG ; Yue WANG ; Yan XU ; Wenhui ZENG ; Wenming LU ; Xingkun TANG ; Wenjie CHEN ; Junsong YE
Chinese Journal of Tissue Engineering Research 2025;29(30):6538-6546
BACKGROUND:In recent years,brain-computer interface technology has shown significant promise for rehabilitating limb dysfunction in stroke patients.With ongoing research deepening and its broader clinical application,combining brain-computer interface with other rehabilitation therapies to improve limb function has become a focal point of study.OBJECTIVE:To analyze and summarize the efficacy of brain-computer interface combined with various therapies in treating limb dysfunction in stroke patients and to explore the clinical value of these combined strategies.METHODS:The search terms used for the literature review in Chinese databases were"brain-computer interface,BCI,stroke,"while the terms"brain-computer interface,BCI,brain-computer interaction,brain-machine interface,BMI,stroke"were used for English databases.Literature searches were conducted in CNKI,WanFang,VIP,PubMed,Embase,and Web of Science,from the time of database construction to September 2024.Finally,a total of 3 054 articles were retrieved,and 75 articles were included after screening for summarization.RESULTS AND CONCLUSION:Currently,brain-computer interfaces,used alone or in combination with other treatments such as Chinese medical treatment,conventional rehabilitation therapy,or physical factor therapy,are achieving better outcomes in treating limb dysfunction in stroke patients.However,the efficacy of brain-computer interfaces combined with transcranial direct current stimulation for treating upper and lower limb dysfunctions is still debated.Researchers are increasingly recognizing the feasibility of these combined therapies.Yet,challenges such as limited exploration of therapeutic mechanisms,absence of standardized protocols,and small sample sizes hinder their broad application.Future research should therefore focus on understanding the mechanisms by which brain-computer interfaces can enhance effects when combined with other therapies and on standardizing criteria for clinical trials to enable widespread clinical adoption.
8.Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury (version 2025)
Aijun XU ; Shuixia LI ; Bo CHEN ; Mengyuan YE ; Lejiao LANG ; Ning NING ; Lin ZHANG ; Changqing LIU ; Zhonglan CHEN ; Weihu MA ; Weishi LI ; Xiaoning WANG ; Dongmei BIAN ; Jiancheng ZENG ; Xin WANG ; Yuan GAO ; Yaping CHEN ; Jiali CHEN ; Yun HAN ; Xiuting LI ; Yang ZHOU ; Xiaojing SU ; Qiong ZHANG ; Tianwen HUANG ; Ping ZHANG ; Hua LIN ; Xingling XIAO ; Ruifeng XU ; Fanghui DONG ; Bing HAN ; Luo FAN ; Yanling PEI ; Suyun LI ; Xiaoju TAN ; Rongchen GUO ; Yefang ZOU ; Xiaoyun HAN ; Junqin DING ; Yi WANG ; Shuhua DENG ; Jinli GUO ; Yinhua LIANG ; Yuan CEN ; Xiaoqin LIU ; Junru CHEN ; Haiyang YU ; Lunlan LI ; Ying REN ; Yunxia LI ; Jianli LU ; Ying YING ; Lan WEI ; Yin WANG ; Qinhong XU ; Yanqin ZHANG ; Yang LYU ; Shijun ZHANG ; Sui WENJIE ; Sanlian HU ; Shuhong YANG ; Guoqing LI ; Jingjing AN ; Baorong HE ; Leling FENG
Chinese Journal of Trauma 2025;41(6):530-541
Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.
9.Effect of "Five-in-One" of traditional Chinese medicine combined with low concentration of atropine in prevention and control of myopia in adolescents
Wenjie YE ; Qiyan ZHANG ; Jianfeng CAO ; Kexin YU
Journal of Clinical Medicine in Practice 2024;28(6):113-117
Objective To explore the safety and effectiveness of the comprehensive therapy of "Five-in-One" of traditional Chinese medicine combined with low concentration of atropine in the prevention and control of myopia in adolescents. Methods Eighty adolescents diagnosed with mild myopia were included in the study. According to the wishes of the subjects and their guardians, they were divided into control group with 40 cases and traditional Chinese medicine intervention group with 40 cases. The control group received 0.01% atropine sulfate eye drops in both eyes once before bedtime every night, the traditional Chinese medicine intervention group underwent twice-weekly comprehensive therapy of "Five-in-One" of traditional Chinese medicine on the basis of local application of medication in the control group. Both groups received a 12-month intervention course. Follow-up visits were conducted before intervention and at 3, 6, and 12 months after treatment. The international standard visual acuity chart was used to examine the changes in unaided distance visual acuity before and after treatment in both groups. The changes of distant visual acuity of naked eye before and after treatment were measured in two groups. The changes in equivalent spherical lens power before and after treatment in both groups were measured by mydriatic refraction combined with a computer optometer. The changes in axial length before and after treatment were measured using an optical biometer. The adverse reactions after treatment in both groups were compared. Results In the control group, the uncorrected distance visual acuity decreased compared with that before intervention as the treatment time prolonged, while was gradually increased in the traditional Chinese medicine intervention group compared with that before intervention (
10.Degranulation of mast cells in renal carcinoma and the correlation with angiogenesis
Leihong YE ; Guodong GUO ; Wenjie YANG ; Yule CHEN
Journal of Modern Urology 2024;29(11):1004-1008
[Objective] To investigate the characteristics of mast cell degranulation in renal carcinoma and the correlation with angiogenesis. [Methods] Mast cells in human renal carcinoma and adjacent tissues were labeled with Tryptase immunohistochemical staining and the infiltration density was calculated.The degranulation level was evaluated according to the distribution of Tryptase inside and outside the mast cells.Vascular endothelial cells were labeled with CD31 immunohistochemical staining and tumor microvascular density was counted.The difference in the number of cells and degranulation level between renal carcinoma and adjacent tissues were compared.The correlations of degranulation with infiltration density and microvessel density were analyzed. [Results] A total of 125 renal carcinoma tissues and 52 adjacent tissues were enrolled in the study.Tryptase staining showed that mast cell density in renal carcinoma and adjacent tissues was (2.67±0.22) and (0.63±0.14) cells/HPF, respectively; the positive rate of mast cell degranulation was 70.3% and 34.3%(P<0.01), respectively.According to the density of mast cells, renal carcinoma tissues were divided into three groups: low, medium and high.The positive rate of mast cell degranulation in the three groups was 42.1%, 64.1% and 83.3%, respectively.Tumor microvessel density in the degranulation positive group was significantly higher than that in the negative group [(24.18±1.64) vs.(10.18±1.97) cells/HPF, P<0.01]. [Conclusion] The level of mast cell degranulation is significantly higher in renal carcinoma than in adjacent tissues, which is associated with infiltration level and angiogenesis.


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