2.Establishment of genomic detection system for Alzheimer′s disease risk based on time-of-flight mass spectrometry
Yuyan KUANG ; Ting ZHANG ; Wenyan GE ; Huimin GUO ; Qingmin RAO ; Yongyin HE ; Qiang WANG ; Xiaomei ZHONG ; Yuping NING ; Yulong LIN ; Haiying LIU
Chinese Journal of Laboratory Medicine 2025;48(12):1571-1580
Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.
3.Establishment of genomic detection system for Alzheimer′s disease risk based on time-of-flight mass spectrometry
Yuyan KUANG ; Ting ZHANG ; Wenyan GE ; Huimin GUO ; Qingmin RAO ; Yongyin HE ; Qiang WANG ; Xiaomei ZHONG ; Yuping NING ; Yulong LIN ; Haiying LIU
Chinese Journal of Laboratory Medicine 2025;48(12):1571-1580
Objective:To establish a genomic nucleic acid mass spectrometry detection platform for allelic risk associated with Alzheimer's disease.Methods:Whole blood samples of 61 patients diagnosed as Alzheimer's disease in the Affiliated Brain Hospital of Guangzhou Medical University from December 28th, 2023 to 31st, March 2024 were collected and deoxynucleic acid (DNA) was extracted, including 22 males and 39 females, aged (67.36 ± 8.18) years old. After screening out 17 risk gene loci in Chinese population, multiplex polymerase chain reaction primers, single-base extension primers and Sanger sequencing primers were designed. Ten samples were used for primer optimization and debugging through Sanger sequencing and time-of-flight mass spectrometry to establish a detection system. The remaining samples were genotyped using a time-of-flight mass spectrometer and verified by Sanger sequencing for accuracy evaluation. Five samples were selected for gradient dilution and then subjected to time-of-flight mass spectrometry detection to evaluate the detection limit. Three clinical samples, one case of Escherichia coli and one case of Staphylococcus aureus genomic DNA samples were selected for cross-reaction research. The anti-interference ability of the detection system was evaluated against hemolysis, chylous substances and conventional anticoagulants in the samples. Two samples, one wild and one homozygous mutation sample with representative peak shapes, were selected to evaluate the anti-interference ability. Four samples containing the common genotypes of all gene loci in the system were selected and repeated 10 times to evaluate the precision.Results:The minimum intensity of single-base extension primers on mass spectrometry is greater than half of the maximum intensity. All 17 risk gene loci screened were successfully typed. The time-of-flight mass spectrometry detection results of 1,037 loci from 61 samples showed that the genotyping detection rate was 100%. The genotypes of the 20 DNA samples were completely consistent with the results of Sanger sequencing, with an accuracy rate of 100%. The mass spectrometry detection results of five samples after gradient dilution indicated that the low detection limit was 5 ng of DNA. The reaction system has a strong anti-interference ability against hemolysis of samples, chylous substances, conventional anticoagulants and DNA cross-contamination. Homologous allele interference and no cross-reaction between the bacterial genome and 17 gene loci do not affect the risk genome detection results. The results of 10 repeated mass spectrometry tests on 4 samples showed that the precision was 100%.Conclusion:The genomic detection system of Alzheimer's disease risk has been successfully established to provide an auxiliary mean for disease diagnosis and risk assessment.
4.Effect of multiparameter electroencephalogram-guided anesthesia management on electroencephalo-gram burst suppression and postoperative delirium in elderly patients undergoing lower abdominal laparoscopic surgery
Jian CHEN ; Yue FENG ; Po SHEN ; Jingjing LIU ; Yi ZHONG ; Xinlong ZHANG ; Jiayong ZHANG ; Yuping HU ; Yanna SI
The Journal of Clinical Anesthesiology 2024;40(9):905-910
Objective To explore the effect of multiparameter electroencephalogram(EEG)-guided anesthesia management on EEG burst suppression(BS)and postoperative delirium(POD)in elderly patients undergoing lower abdominal laparoscopic surgery.Methods A total of 100 elderly patients,48 males and 52 females,aged 65-85 years,BMI 18.5-28.0 kg/m2,and ASA physical status Ⅱ or Ⅲ,were enrolled for lower abdominal surgery under general anesthesia.Patients were randomly divided into two groups:multiparameter group and single parameter group,50 patients in each group.In multiparameter group,multiparameter EEG monitoring with patient statu index(PSI),spectral edge frequency(SEF),burst suppression ratio(BSR)and density spectral array(DSA)were used to guide the depth management of anesthesia.In single parameter group,single parameter PSI was used to guide the depth management of anesthesia.The total area under the hypotensive threshold of MAP(AUTMAP)was calculated,and the amount of anesthetic used during the operation and the use of vasoactive drugs,duration of anesthesia,extu-bation time,duration of PACU stay,and postoperative hospitalisation days were recorded.HR,MAP,PSI,and SEF were recorded before the induction of anesthesia,5 minutes after induction of anesthesia,5,30,and 60 minutes after incision,and at the end of surgery.The incidence,duration,and maximum BSR of in-traoperative BS,as well as the incidence of POD 1,2,and 3 days after surgery were recorded.Results There was no significant difference in AUTMAP values between the two groups.Compared with single parame-ter group,intraoperative propofol and remifentanil dosage were significantly decreased(P<0.05),awak-ening time,PACU stay,and postoperative hospitalization time were significantly shorter in multiparameter group(P<0.05),the PSI was significantly increased 5,30,and 60 minutes after incision and at the end of surgery,and the SEF was significantly increased 5 minutes after induction of anesthesia,5,30,and 60 minutes after induction and the end of surgery(P<0.05).Compared with single parameter group,inci-dence of intraoperative BS was significantly decreased,duration of BS was significantly shorter,smaller maximum BSR was significantly decreased,and incidence of POD on 1 day after surgery in multiparameter group(P<0.05).Conclusion Anesthesia management guided by multiparameter EEG can inhibit the oc-currence of BS,mitigate the degree of BS,and reduce the incidence of POD in elderly patients undergoing abdominal surgery.
5.Impact of nutrition education on vitamin D nutritional status in pregnant women
Yuping ZHANG ; Rui LI ; Xiaoqin ZHONG ; Yanping LIU
Chinese Journal of Health Management 2024;18(11):824-829
Objective:To explore the impact of nutrition education on the vitamin D nutritional status of pregnant women.Methods:It was a randomized controlled trial. A total of 96 women with vitamin D deficiency at 8-13 weeks of gestation who were regularly followed-up in the pregnancy nutrition clinic of the Department of Clinical Nutrition of Peking Union Medical College Hospital from January to October 2021 were selected by convenience sampling. The women were divided into a control group and an experimental group (48 cases in each group) according to the random number table method. The control group received routine outpatient treatment and was given different doses of vitamin D supplements according to the degree of vitamin D deficiency. On this basis, the experimental group were given additional nutritional education, including the distribution of vitamin D education manuals at 8-13 weeks of pregnancy, the assessment of vitamin D intake and cognition of vitamin D, the provision of individualized vitamin D nutritional status improvement plans, and the one-time assessment and guidance of vitamin D nutritional status at 24-28 weeks and 36-38 weeks of pregnancy, to continuously promote vitamin D health. Due to factors such as miscarriage or incomplete data acquisition during the study, 44 cases were finally included in the control group and 42 cases in the experimental group. The non-parametric test Mann-Whitney U was used to compare the vitamin D nutritional status of the two groups at 24-28 weeks and 36-38 weeks of pregnancy, and to analyze the effect of nutritional education on the vitamin D nutritional status of pregnant women. Results:At 8-13 weeks of pregnancy, there was no significant difference in serum 25-hydroxyvitamin D [25(OH)D] concentration between the two groups ( P>0.05). At 24-28 weeks of pregnancy, the serum 25(OH)D concentration and the proportion of pregnant women with normal vitamin D nutritional status in the experimental group were both significantly higher than those in the control group [29.90 (24.63, 34.28) vs 25.65 (21.50, 31.28) μg/L, 50.0% vs 29.5%] (both P<0.05). At 36-38 weeks of pregnancy, there was no significant difference in serum 25(OH)D level between the two groups ( P>0.05); however, the proportion of pregnant women with normal vitamin D nutritional status in the control group and the experimental group was 80.5% and 70.7%, respectively, which were both significantly higher than those in 8-13 and 24-28 weeks of pregnancy. The main effect of time was significant in both groups ( F=201.040, P<0.001). Conclusion:Nutrition education can enhance pregnant women′s knowledge of vitamin D, promote behavioral changes, and improve vitamin D nutritional status.
6.Efficacy and safety of bendamustine-rituximab combination therapy for newly diagnosed indolent B-cell non-Hodgkin's lymphoma and elderly mantle cell lymphoma: a multi-center prospective phase II clinical trial in China
Hui WANG ; Qiang HE ; Dan LIU ; Xiuzhi DENG ; Ji MA ; Linna XIE ; Zhongliang SUN ; Cong LIU ; Rongrong ZHAO ; Ke LU ; Xiaoxia CHU ; Na GAO ; Haichen WEI ; Yanhua SUN ; Yuping ZHONG ; Lijie XING ; Haiyan ZHANG ; Hao ZHANG ; Wenwei XU ; Zengjun LI
Chinese Journal of Hematology 2023;44(7):550-554
Objectives:This study aimed to assess the efficacy and safety of bendamustine in combination with rituximab (BR regimen) for the treatment of newly diagnosed indolent B-cell non-Hodgkin's lymphoma (B-iNHL) and elderly mantle cell lymphoma (eMCL) .Methods:From December 1, 2020 to September 10, 2022, a multi-center prospective study was conducted across ten Grade A tertiary hospitals in Shandong Province, China. The BR regimen was administered to evaluate its efficacy and safety in newly diagnosed B-iNHL and eMCL patients, and all completed at least four cycles of induction therapy.Results:The 72 enrolled patients with B-iNHL or MCL were aged 24-74 years, with a median age of 55 years. Eastern Cooperative Oncology Group (ECOG) performance status scores of 0-1 were observed in 76.4% of patients, while 23.6% had scores of 2. Disease distribution included follicular lymphoma (FL) (51.4% ), marginal zone lymphoma (MZL) (33.3% ), eMCL (11.1% ), and the unknown subtype (4.2% ). According to the Ann Arbor staging system, 16.7% and 65.3% of patients were diagnosed with stage Ⅲ and stage Ⅳ lymphomas, respectively. Following four cycles of BR induction therapy, the overall response rate was 98.6%, with a complete response (CR) rate of 83.3% and a partial response (PR) rate of 15.3%. Only one eMCL patient experienced disease progression during treatment, and only one FL patient experienced a relapse. Even when evaluated using CT alone, the CR rate was 63.9%, considering the differences between PET/CT and CT assessments. The median follow-up duration was 11 months (range: 4-22), with a PFS rate of 96.8% and an OS rate of 100.0%. The main hematologic adverse reactions included grade 3-4 leukopenia (27.8%, with febrile neutropenia observed in 8.3% of patients), grade 3-4 lymphopenia (23.6% ), grade 3-4 anemia (5.6% ), and grade 3-4 thrombocytopenia (4.2% ). The main non-hematologic adverse reactions such as fatigue, nausea/vomiting, rash, and infections occurred in less than 20.0% of patients.Conclusion:Within the scope of this clinical trial conducted in China, the BR regimen demonstrated efficacy and safety in treating newly diagnosed B-iNHL and eMCL patients.
7.Research progress on transcranial alternating current stimulation for cognitive impairment in the elderly
Zhangying WU ; Xiaomei ZHONG ; Yuping NING
Chinese Journal of Psychiatry 2023;56(5):342-347
Transcranial alternating current stimulation (tACS) can modulate human neural activity and behavior by inducing local changes in neural oscillatory power and cross-frequency, cross-regional phase synchronization. It has enormous potential for cognitive research and brain condition treatment. Cognitive impairment in the elderly involves dysrhythmic neural activity within and between brain regions. tACS regulates single or nested brain rhythms to induce cell synchronization and desynchronization, enhancing local or distant synchronous activity in the cortical network to improve cognitive function. This review focuses on introducing the possible mechanisms, influencing factors of efficacy, and clinical applications for elderly cognitive impairment, as well as the development prospects of cross-frequency coupling and closed-loop modulation and other paradigms.
8.Clinical treatment guideline for pulmonary blast injury (version 2023)
Zhiming SONG ; Junhua GUO ; Jianming CHEN ; Jing ZHONG ; Yan DOU ; Jiarong MENG ; Guomin ZHANG ; Guodong LIU ; Huaping LIANG ; Hezhong CHEN ; Shuogui XU ; Yufeng ZHANG ; Zhinong WANG ; Daixing ZHONG ; Tao JIANG ; Zhiqiang XUE ; Feihu ZHOU ; Zhixin LIANG ; Yang LIU ; Xu WU ; Kaican CAI ; Yi SHEN ; Yong SONG ; Xiaoli YUAN ; Enwu XU ; Yifeng ZHENG ; Shumin WANG ; Erping XI ; Shengsheng YANG ; Wenke CAI ; Yu CHEN ; Qingxin LI ; Zhiqiang ZOU ; Chang SU ; Hongwei SHANG ; Jiangxing XU ; Yongjing LIU ; Qianjin WANG ; Xiaodong WEI ; Guoan XU ; Gaofeng LIU ; Junhui LUO ; Qinghua LI ; Bin SONG ; Ming GUO ; Chen HUANG ; Xunyu XU ; Yuanrong TU ; Liling ZHENG ; Mingke DUAN ; Renping WAN ; Tengbo YU ; Hai YU ; Yanmei ZHAO ; Yuping WEI ; Jin ZHANG ; Hua GUO ; Jianxin JIANG ; Lianyang ZHANG ; Yunfeng YI
Chinese Journal of Trauma 2023;39(12):1057-1069
Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.
9.Research progress on transcranial alternating current stimulation for cognitive impairment in the elderly
Zhangying WU ; Xiaomei ZHONG ; Yuping NING
Chinese Journal of Psychiatry 2023;56(5):342-347
Transcranial alternating current stimulation (tACS) can modulate human neural activity and behavior by inducing local changes in neural oscillatory power and cross-frequency, cross-regional phase synchronization. It has enormous potential for cognitive research and brain condition treatment. Cognitive impairment in the elderly involves dysrhythmic neural activity within and between brain regions. tACS regulates single or nested brain rhythms to induce cell synchronization and desynchronization, enhancing local or distant synchronous activity in the cortical network to improve cognitive function. This review focuses on introducing the possible mechanisms, influencing factors of efficacy, and clinical applications for elderly cognitive impairment, as well as the development prospects of cross-frequency coupling and closed-loop modulation and other paradigms.
10.Metformin inhibits apoptosis by regulating TET2-Foxo3a pathway after spinal cord injury
Jiwei ZHAO ; Zhigang MIAO ; Huihui SUN ; Le HU ; Hao SUN ; Xiaoli ZHONG ; Xinmin FENG ; Jiandong YANG ; Yuping TAO ; Jun CAI ; Liang ZHANG ; Jingcheng WANG ; Yongxiang WANG
Chinese Journal of Orthopaedics 2021;41(9):584-594
Objective:Through TTC staining, immunohistochemical analysis, RT-PCR and hind limb motor function evaluation and other experimental methods, to explore the regulatory mechanism of metformin on anti-apoptosis in rats with spinal cord injury (SCI).Methods:Establish a rat spinal cord injury model. Through Basso-Beattie -Bresnahan locomotor rating scale (BBB) and cant test to evaluate the recovery of hindlimb motor function in rats. The changes of necrotic area of spinal cord tissue were compared by TTC staining. Extraction of rat spinal cord tissue, by Dot blot analysis and immunohistochemical detection of the hydroxyl of DNA methylation level. By qPCR, Western Blot detection TET2mRNA and protein expression level, and the changes in the scope of spinal cord injury were detected by inhibiting the expression of TET2. The interaction between TET2 and Foxo3a was detected by immunoblotting and immunoprecipitation. Through RT-PCR assay Foxo3a downstream related changes in the level of gene expression.Results:Compared with the SCI+NS group, the necrotic area of the spinal cord tissue was reduced after metformin treatment, and the BBB score and the incline test score were higher ( P<0.05). At the same time, we found that the levels of TET2mRNA and protein increased significantly after SCI at 24 h, and the 5-hmC level of DNA increased. The levels of TET2mRNA and protein and 5-hmC increased further after the use of metformin. After using SC-1, compared with the SCI+MET group, the level of 5-hmC decreased and the area of infarction increased. After SCI, the mRNA levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the mRNA levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). After SCI, the 5-hmC levels of downstream genes Bim, P27kip, Bax increased significantly. After metformin treatment, the 5-hmC levels of Bim and Bax were lower than those in the SCI+NS group ( P<0.05). Conclusion:Metformin can promote the interaction between TET2 and Foxo3a, increase the 5-hmC level of the overall DNA, and inhibit the activation of related apoptosis genes, thereby improving tissue damage and nerve function recovery after spinal cord injury.

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