1.Trace component fishing strategy based on offline two-dimensional liquid chromatography combined with PRDX3-surface plasmon resonance for Uncaria alkaloids.
Hui NI ; Zijia ZHANG ; Ye LU ; Yaowen LIU ; Yang ZHOU ; Wenyong WU ; Xinqin KONG ; Liling SHEN ; Sihan CHEN ; Huali LONG ; Cheng LUO ; Hao ZHANG ; Jinjun HOU ; Wanying WU
Journal of Pharmaceutical Analysis 2025;15(9):101244-101244
The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C18 column and a conventional C18 column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H2O2 measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H2O2 clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field.
2.Characterization of hemolytic transfusion reactions induced by anti-E antibodies
Huali HUANG ; Hao LI ; Yuerong WEI ; Ruixian LUO ; Huini HUANG ; Huiqiong XIE ; Hailan LI ; Ziji YANG ; Zhuning MO
Chinese Journal of Blood Transfusion 2025;38(11):1577-1585
Objective: To investigate the causes and characteristics of immune hemolytic transfusion reactions (HTRs) triggered by anti-E antibodies, so as to provide a scientific reference for guaranteeing clinical blood transfusion safety. Methods: Five patients who experienced HTRs in our hospital from November 2023 to October 2024 were selected as the research subjects. ABO/RhD blood grouping, antibody screening, antibody identification, and the direct antiglobulin test (DAT) were conducted using the column agglutination method. The causes of HTRs in these patients were investigated using multiple techniques such as the two-step enzyme method, polyethylene glycol (PEG), acid elution technique, and capillary centrifugation method. Results: All five patients tested negative for antibody screening prior to transfusion. However, after transfusion of E+ phenotyped blood, patients 1, 2, 3, and 5 developed delayed haemolytic transfusion reaction (DHTR), while patient 4 experienced acute haemolytic transfusion reaction (AHTR). Anti-E antibodies were detected in all blood samples from the patients after the hemolytic transfusion reaction, including the enzyme-only anti-E antibody in two cases. Conclusion: Anti-E antibody can trigger both intravascular and extravascular hemolysis. It is recommended to conduct ABO/RhD and RhE antigen-matched transfusions and establish a regional blood transfusion database to reduce immune hemolytic transfusion reactions caused by anti-E antibody.
3.Genome-wide DNA methylation and mRNA transcription analysis revealed aberrant gene regulation pathways in patients with dermatomyositis and polymyositis.
Hui LUO ; Honglin ZHU ; Ding BAO ; Yizhi XIAO ; Bin ZHOU ; Gong XIAO ; Lihua ZHANG ; Siming GAO ; Liya LI ; Yangtengyu LIU ; Di LIU ; Junjiao WU ; Qiming MENG ; Meng MENG ; Tao CHEN ; Xiaoxia ZUO ; Quanzhen LI ; Huali ZHANG
Chinese Medical Journal 2025;138(1):120-122
4.Antibody-based gene expression profiling of peripheral blood from patients with idiopathic inflammatory myopathy
Muyuan LI ; Li WANG ; Quanzhen LI ; Hui LUO ; Huali ZHANG
Chinese Journal of Rheumatology 2024;28(1):15-21
Objective:To elucidate the pathophysiological mechanisms of idiopathic inflammatory myopathy subtypes by analyzing the gene expression profiles of peripheral blood mononuclear cells (PBMCs) from anti-MDA5 antibody-positive and anti-Jo-1 antibody-positive myositis patients.Methods:Gene expression profiling screening and analysis of PBMCs from 12 anti-MDA5 positive, 16 anti-Jo-1 positive myositis patients and 43 healthy controls were performed using Illumina HT-12 v4 expression profiling microarrays. Applying the unpaired t test with Benjamini-Hochberg correction, the genes with the absolute value of fold change (FC) in gene expression signal ≥2 and adjusted P<0.05 were selected as differentially expressed genes. Differential gene sets were subjected to Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, with P<0.05 as the threshold for being significantly enriched. Validation of differentially expressed genes by real time-PCR. The Kolmogorov-Smirnov test was used to test the normality of continuous variables. If the distribution was normal and the variance was homogeneous, analysis of variance (one-way ANOVA) was used.If the distribution was not normal, Kruskal-Wallis test was used, and P<0.05 was regarded as statistically significant difference. Results:Analysis of gene expression profiles of PBMCs from patients with positive anti-MDA5 and anti-Jo-1 antibody revealed significant differences in gene expression of PBMCs from patients with the two myositis subtypes. The number of differentially expressed genes that specifically up-regulated in anti-MDA5 antibody positive patients was 407, and the GO functional enrichment analysis was mainly enriched in biological processes such as innate immune response ( P<0.001), response to virus ( P<0.001) and type Ⅰ interferon signaling pathway ( P<0.001), and the KEGG pathway enrichment analysis was mainly enriched in the viral infection-associated pathway ( P<0.001), RIG-Ⅰ like receptor signaling pathway ( P<0.001) and Toll-like receptor signaling pathway ( P=0.002), etc. The 259 differential genes specifically down-regulated in the anti-MDA5 antibody positive group were mainly enriched in biological processes such as immune response ( P=0.006), TGF-β receptor signaling pathway ( P=0.010) and natural killer cell mediated immunity ( P=0.015) in GO functional enrichment analysis. There were 162 differentially expressed genes up-regulated specifically in anti-Jo-1 antibody positive patients, and GO functional enrichment analysis was mainly enriched in biological processes such as nucleosome assembly ( P<0.001), negative regulation of cell growth ( P=0.001), negative regulation of apoptotic process P=0.004), and innate immune response in mucosa ( P=0.012), and the KEGG pathway enrichment analysis mainly enriched in metabolic-related signaling pathways ( P<0.001) and immune-related pathways ( P<0.001), etc. Real-time PCR confirmed that IFIH1 ( P=0.037), ISG15 ( P=0.003), and DDX58 ( P=0.032) in the RIG-Ⅰ-like receptor pathway as well as chemokines MCP-1 ( P=0.003), MCP-2 ( P<0.001), and transcription factor BATF2 ( P=0.002), and inflammatory signaling pathway-associated MYD88 ( P<0.001) were highly expressed in PBMCs from anti-MDA5 antibody-positive myositis patients. Conclusion:The gene expression profile of PBMCs in anti-MDA5 antibody-positive patients suggests that the pathogenesis of patients with anti-MDA 5 antibody positive is closely related to biological processes such as innate immune response, viral infection, and interferon response.
5.The Role of Nrf2 in Exercise Improving of NAFLD
Ge ZHAO ; Yuan LUO ; Ya-Ping LI ; Yan-Qing YAN ; Shu-Jing LIU
Progress in Biochemistry and Biophysics 2024;51(5):1079-1089
In cardiovascular disorders, neurological diseases, and chronic metabolic diseases, the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway is essential for maintaining cell homeostasis. According to studies, boosting Nrf2 expression can be used to cure or prevent chronic diseases that are characterized by oxidative stress, inflammation, and mitochondrial dysfunction. Nonalcoholic fatty liver disease (NAFLD) is a chronic metabolic liver disease characterized by hepatic steatosis brought on by a number of causes other than alcohol. In recent years, its incidence has gradually risen across the globe. According to relevant studies, NAFLD and the Nrf2 signaling pathway are tightly connected. Inhibiting lipid production and metabolism-related enzymes, repairing impaired liver metabolism, and lowering hepatic lipid storage are all possible with Nrf2 activation. Exercise is a powerful tool for treating and preventing NAFLD. However, exercise type, exercise intensity, environment, and exhaustion all have an impact on the Nrf2 signaling pathway. By activating Nrf2, exercise can lessen liver inflammation, oxidative stress, endoplasmic reticulum stress, and insulin resistance, and ameliorate liver damage to improve NAFLD. The activation of Nrf2 signaling pathway, its associated mechanism of controlling antioxidation, and the impact of exercise on the Nrf2 signaling pathway are all explained in this work. Based on the pathogenesis of NAFLD, this article examines the connection between exercise, Nrf2, and NAFLD, and the current state of knowledge regarding Nrf2’s role in the amelioration of NAFLD through exercise. It offers a theoretical frame of reference for future research into how Nrf2 might be used to improve NAFLD.
6.Polyarthritis, hyperuricemia and right shoulder pain: a report of one case with literature review
Tao LUO ; Huali LUO ; Yuanchun MO ; Qing YIN ; Yingzhu CHEN
Chinese Journal of Rheumatology 2023;27(1):22-27,C1-4
Objective:To improve the clinical differential diagnosis ability of rheumatoid arthritis (RA) complicated with gout and septic arthritis (SA).Methods:The clinical characteristics, diagnosis, and treatment of one RA patient with hyperuricemia and recurrent swelling and pain in right shoulder were reported and discussed.Results:A patient, with a history of RA for 10 years, hyperuricemia for 8 years, recurrent swelling and pain in right shoulder for 1 year. RA, gout, and SA were diagnosed before, and the response was poor after symptomatic treatment. In recent 1 month, the symptom was aggravated with the formation of fistula on the right shoulder. The laboratory tests for tuberculosis T cell interferon release test (IGRA) and tuberculin (PPD) test were negative, and the CD4 + cell count decreased. The comprehensive analysis of the imaging with right shoulder showed MSU deposition on right shoulder, with bone erosion, bone destruction, bone marrow edema, joint effusion, and multiple sites of connective tissue involvement (synovial bursa, tendon sheath, tendon, and muscle) GeneXpert MTB/RIF (GeneXpert), metagenomic next-generation sequencing (mNGS) of puncture fluid and joint fluid culture prompted Mycobacterium tuberculosis complex group. He was finally diagnosed with RA, gout, and osteoarticular tuberculosis (OAT). Symptoms were relieved after symptomatic treatment. Conclusion:RA patients with hyperuricemia have recurrent single arthritis. In addition to considering for gout, the presence of OAT should also be considered. The immune functional status of the patient and drug used may interfere with the interpretation of immune function tests. It is necessary to integrate the clinical characteristics of patients, a variety of imaging examinations, and etiological detection to confirm the diagnosis and avoid misdiagnosis.
7.Effects of miRNA-628-3p on proliferation, apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor
Huali PENG ; Tongchen HU ; Fan YANG ; Fan ZHANG ; Wei LIU ; Wei LUO
Cancer Research and Clinic 2022;34(7):481-486
Objective:To investigate the effects of miRNA-628-3p (miR-628-3p) on the proliferation, apoptosis and invasion of non-small cell lung cancer H1299 cells and its targeting relationship with insulin-like growth factor 1 receptor (IGF-1R).Methods:The blank control group (untreated H1299 cells), miR-NC group (H1299 cells transfected with empty plasmid), miR-628-3p-M group (H1299 cells transfected with miR-628-3p mimic sequence plasmid) and miR-628-3p-I group (H1299 cells transfected with miR-628-3p inhibitory sequence plasmid) were established. The cells in each group were cultured for 72 h, and the cell proliferation ability was detected by methyl thiazol tetrazolium (MTT) method, the number of cell monoclonal formation was determined by crystal violet staining, the level of cell apoptosis was determined by flow cytometry, and the cell invasion ability was determined by Transwell method. The mRNA levels of miR-628-3p and IGF-1R in cells were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the protein level of IGF-1R in cells was determined by Western blotting.Results:Compared with the blank control group and miR-NC group, the cell survival rate [(42±7)% vs. (78±6)%, (76±7)%], the number of monoclonal formation [235±35 vs. 614±89, 618±75], the number of invasive cells [(265±85) cells vs. (693±185) cells, (703±119) cells], relative expression of IGF-1R mRNA (2.17±0.14 vs. 3.38±0.15, 3.37±0.13) and relative expression of IGF-1R protein (0.34±0.13 vs. 0.89±0.19, 0.88±0.18) in the miR-628-3p-M group were lower (all P < 0.05), but the apoptosis rate [(9.30±3.51)% vs. (3.30±1.54)%, (3.10±1.94)%] and relative expression of miR-628-3p (6.93±0.17 vs. 3.29±0.15, 3.30±0.16) were higher (all P < 0.05); the cell survival rate [(90±6)%], the number of monoclonal formation (1 063±102), the number of invasive cells [(1 985±426) cells], relative expression of IGF-1R mRNA (4.30±0.18) and relative expression of IGF-1R protein (1.47±0.17) in the miR-628-3p-I group were higher (all P < 0.05), but the apoptosis rate [(0.90±0.20)%] and the relative expression of miR-628-3p (1.93±0.18) were lower (both P < 0.05). Compared with the miR-628-3p-M group, the miR-628-3p-I group had higher cell survival rate, the number of monoclonal formation, the number of invasive cells, and the relative expressions of IGF-1R mRNA and protein (all P < 0.05), but the apoptosis rate and relative expression of miR-628-3p were lower (both P < 0.05). Conclusions:After regulation of miR-628-3p level, the proliferation, migration and invasion of H1299 cells are affected. miR-628-3p may have a targeting relationship with IGF-1R.
8.Cognitive impairment and psychotic symptoms in patients with general paresis of insane: a follow-up study
Ben CHEN ; Haishan SHI ; Xiaomei ZHONG ; Le HOU ; Huali WANG ; Yanhua WANG ; Xinru CHEN ; Xinni LUO ; Zhangying WU ; Qi PENG ; Lijuan LI ; Yuping NING
Chinese Journal of Psychiatry 2017;50(3):226-230
Objective To investigate the characteristics of cognitive impairment and psychotic symptoms in general paresis of insane (GPI) before and after penicillin therapy, and explore factors that may predict the clinical outcomes. Methods Thirty patients with GPI were recruited. All GPI patients underwent a comprehensive neuropsychological assessment before receiving penicillin therapy, and returned for follow-up visits after 7 months. The severity of dementia was determined by Clinical Dementia Rating Scale (CDR), cognitive functions were assessed by Mini Mental State Examination (MMSE) and Alzheimer 's Disease Assessment Scale-cognitive subscale (ADAS-Cog), ability of daily living was assessed by Instrumental Activities of Daily Living Scale (IALD) and Physical Self maintenance Scale(PSMS), behavioral and psychological symptoms were assessed by Neuropsychiatric Inventory (NPI). Aqueous crystalline penicillin G 24 million units per day was administered as continuous infusion for 14 days, followed by benzathine penicillin 2.4 million units IM once per week for 3 weeks. Patients returned for follow-up visits after 7 months. Clinical outcomes were determined by the improvement of neuropsychological test scores at the end of the treatment. Grouped by CDR scores, changes in neuropsychological tests scores among different GPI groups were used to explore the correlation between severity of dementia and clinical outcomes. Univariate analysis and multivariate linear regression analysis were used to identify factors that may predict the clinical outcomes. Results (1)After penicillin therapy, GPI patients' MMSE scores(14.4± 6.9 vs.17.1 ± 9.1)and IADL scores(4.0(2.0, 5.0)vs.6.0(2.0, 7.3))both improved significantly(t=5.820, Z=3.710, P<0.01),while in ADAS-Cog, only factor scores of attention(1.5(0.7, 3.0)vs.1.5(0, 2.3))reduced significantly(Z=- 2.680, P<0.01). NPI's total scores(46.0 ± 27.1 vs.17.6 ± 15.4)and subscores of hallucination, delusion, agitation, depression, euphoria, disinhibition and irritability reduced significantly (Z=-4.940,-2.381,-2.504,-3.095,-2.492,-3.097,-2.527,-3.715, all P<0.05).(2) Grouped by the CDR scores, MMSE scores and IADL scores in very mild GPI group with CDR=0.5 improved significantly. In mild GPI group with CDR=1, significant changes were also found in all neuropsychological tests scores(MMSE,t=5.409, P<0.01), total scores of ADAS-Cog (Z=-2.366,P<0.05), IADL (Z=2.546, P<0.05), total scores of NPI (Z=-3.558,P<0.01), but except for PSMS. In moderate to severe GPI group with CDR>1,significant change was found only in total scores of NPI (t=-3.772,P<0.05). (3) Univariate analysis and multivariate linear regression analysis showed that improvement of MMSE scores after the treatment was significantly correlated with IADL scores and MMSE scores at baseline(β=0.541,P=0.004;β=0.364,P=0.044). Conclusions After penicillin treatment, GPI patients may improve in both cognitive function and psychotic symptoms but not in all the domains. Symptoms of anxiety, sleep/nigh-time behavior change, and apathy, as well as moderate to severe GPI patients may not benefit much from the treatment.
9.Cognitive impairment and psychotic symptoms in patients with general paresis of insane: a follow-up study
Ben CHEN ; Haishan SHI ; Xiaomei ZHONG ; Le HOU ; Huali WANG ; Yanhua WANG ; Xinru CHEN ; Xinni LUO ; Zhangying WU ; Qi PENG ; Lijuan LI ; Yuping NING
Chinese Journal of Psychiatry 2017;50(3):226-230
Objective To investigate the characteristics of cognitive impairment and psychotic symptoms in general paresis of insane (GPI) before and after penicillin therapy, and explore factors that may predict the clinical outcomes. Methods Thirty patients with GPI were recruited. All GPI patients underwent a comprehensive neuropsychological assessment before receiving penicillin therapy, and returned for follow-up visits after 7 months. The severity of dementia was determined by Clinical Dementia Rating Scale (CDR), cognitive functions were assessed by Mini Mental State Examination (MMSE) and Alzheimer 's Disease Assessment Scale-cognitive subscale (ADAS-Cog), ability of daily living was assessed by Instrumental Activities of Daily Living Scale (IALD) and Physical Self maintenance Scale(PSMS), behavioral and psychological symptoms were assessed by Neuropsychiatric Inventory (NPI). Aqueous crystalline penicillin G 24 million units per day was administered as continuous infusion for 14 days, followed by benzathine penicillin 2.4 million units IM once per week for 3 weeks. Patients returned for follow-up visits after 7 months. Clinical outcomes were determined by the improvement of neuropsychological test scores at the end of the treatment. Grouped by CDR scores, changes in neuropsychological tests scores among different GPI groups were used to explore the correlation between severity of dementia and clinical outcomes. Univariate analysis and multivariate linear regression analysis were used to identify factors that may predict the clinical outcomes. Results (1)After penicillin therapy, GPI patients' MMSE scores(14.4± 6.9 vs.17.1 ± 9.1)and IADL scores(4.0(2.0, 5.0)vs.6.0(2.0, 7.3))both improved significantly(t=5.820, Z=3.710, P<0.01),while in ADAS-Cog, only factor scores of attention(1.5(0.7, 3.0)vs.1.5(0, 2.3))reduced significantly(Z=- 2.680, P<0.01). NPI's total scores(46.0 ± 27.1 vs.17.6 ± 15.4)and subscores of hallucination, delusion, agitation, depression, euphoria, disinhibition and irritability reduced significantly (Z=-4.940,-2.381,-2.504,-3.095,-2.492,-3.097,-2.527,-3.715, all P<0.05).(2) Grouped by the CDR scores, MMSE scores and IADL scores in very mild GPI group with CDR=0.5 improved significantly. In mild GPI group with CDR=1, significant changes were also found in all neuropsychological tests scores(MMSE,t=5.409, P<0.01), total scores of ADAS-Cog (Z=-2.366,P<0.05), IADL (Z=2.546, P<0.05), total scores of NPI (Z=-3.558,P<0.01), but except for PSMS. In moderate to severe GPI group with CDR>1,significant change was found only in total scores of NPI (t=-3.772,P<0.05). (3) Univariate analysis and multivariate linear regression analysis showed that improvement of MMSE scores after the treatment was significantly correlated with IADL scores and MMSE scores at baseline(β=0.541,P=0.004;β=0.364,P=0.044). Conclusions After penicillin treatment, GPI patients may improve in both cognitive function and psychotic symptoms but not in all the domains. Symptoms of anxiety, sleep/nigh-time behavior change, and apathy, as well as moderate to severe GPI patients may not benefit much from the treatment.
10.Effect of psychological intervention in the rectal cancer after operation
Mindan CHEN ; Xiaohui JIN ; Ya CHEN ; Xiufang CHEN ; Minchang GUAN ; Huali LUO ; Jinlin DU
China Modern Doctor 2015;(21):140-142,146
Objective To study the effect of psychological intervention in the rectal cancer after operation. Methods Thirty-seven patients each in control group and intervention group with rectal cancer after operation from February 2012 to February 2015 in our department were retrospectively analyzed. The complications, quality of life and psycho-logical status were compared between the two groups. Results The intervention group was significantly lower than that of the control group in urinary tract infection (χ2=4.44, P<0.05), intestinal obstruction (χ2=3.85, P<0.05), pulmonary infection(χ2=6.51, P<0.05). The intervention group were significantly higher than that in the control group after 4 weeks of the operation in physical function(t=5.75, P<0.01), role function(t=5.50, P<0.01), emotional function(t=6.08, P<0.01), cognitive function(t=6.60, P<0.01), social function(t=2.92, P<0.01). The intervention group was significantly lower than that of the control group in SAS(t=4.05, P<0.01) and SDS (t=3.11, P<0.01). Conclusion Psychological inter-vention can decrease the rate of postoperative complications and improve the psychological state in the rectal cancer after operation, it can also enhance the quality of life.

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