1.Mechanism of Xiezhuo Jiedu Formula in Treating Ulcerative Colitis Through Pyroptosis Regulation Based on Bioinformatics and Animal Experiments
Qiang CHUAI ; Wenjing ZHAI ; Shijie REN ; Xiaomeng LANG ; Xin KANG ; Wenli WEI ; Jingyuan LIU ; Jianping LIU ; Jie REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):105-113
ObjectiveThis study aims to explore the potential mechanism of the Xiezhuo Jiedu formula in regulating pyroptosis for the treatment of ulcerative colitis (UC) using bioinformatics and in vivo animal experiments. MethodsDifferentially expressed genes (DEGs) in colon tissues of UC patients were retrieved from the Gene Expression Omnibus (GEO) database. Pyroptosis-related genes were obtained from the GEO and GeneCards databases. The intersection of these datasets yielded pyroptosis-related DEGs (Pyro-DEGs). Pyro-DEGs were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis using the Metascape database. A protein-protein interaction (PPI) network was constructed using the STRING database. Least absolute shrinkage and selection operator (LASSO) prediction model and receiver operating characteristic (ROC) analysis were conducted to identify core Pyro-DEGs with diagnostic and therapeutic potential. Immune infiltration analysis of the UC datasets was performed using the deconvolution method (CIBERSORT), along with correlation analysis with core Pyro-DEGs. Sixty male Sprague-Dawley (SD) rats were randomly divided into a control group, a model group, high-, medium-, and low-dose groups of Xiezhuo Jiedu formula (26.64, 13.32, 6.66 g·kg-1), and a mesalazine group (0.27 g·kg-1), with 10 rats in each group. UC was established by intrarectal administration of 3,5-trinitrobenzenesulfonic acid (TNBS) dissolved in ethanol. The control and model groups were given distilled water by gavage, while the treatment groups were administered the corresponding drugs for 7 consecutive days. Hematoxylin-eosin (HE) staining was used to observe the colon histopathology. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors such as interleukin-1β (IL-1β), IL-10, IL-18, and transforming growth factor-β (TGF-β). Immunohistochemistry (IHC) and Western blot were applied to detect the expression of Caspase-1, gap junction alpha-1 protein (GJA1), peroxisome proliferator-activated receptor gamma (PPARG), and S100 calcium-binding protein A8 (S100A8). Real-time quantitative polymerase chain reaction (Real-time PCR) was utilized to measure mRNA expression of Caspase-1, GJA1, PPARG, and S100A8. Western blot was performed to assess protein expression levels of Caspase-1, GJA1, PPARG, and S100A8. ResultsGEO datasets GSE87466 and GSE87473 yielded 64 Pyro-DEGs. KEGG analysis indicated that these genes were enriched in the NOD-like receptor signaling pathway, tumor necrosis factor (TNF) signaling pathway, and hypoxia-inducible factor 1 (HIF-1) signaling pathway. Four core Pyro-DEGs (Caspase-1, GJA1, PPARG, and S100A8) were identified. Immune infiltration analysis showed that expression of these genes was positively correlated with mast cells, neutrophils, M0 macrophages, M1 macrophages, and dendritic cells. Animal experimental results indicated that compared with the control group, the model group had significantly increased levels of IL-1β and IL-18, significantly decreased levels of IL-10 and TGF-β. The model group showed enhanced Caspase-1, GJA1, and S100A8 staining, and significantly increased mRNA and protein expression of Caspase-1, GJA1, and S100A8 (P<0.01). In contrast, the expression of PPARG was reduced in the model group (P<0.01). After treatment, all dosage groups showed varying degrees of improvement (P<0.05, P<0.01), with the high-dose group showing the most significant improvement (P<0.01). ConclusionCaspase-1, GJA1, PPARG, and S100A8 are core Pyro-DEGs closely associated with the pathogenesis of UC. These genes may collaborate with immune cells such as mast cells, neutrophils, and M0 macrophages to mediate disease development. The Xiezhuo Jiedu formula may regulate the expression of core Pyro-DEGs through the NOD-like receptor, TNF, and HIF-1 core signaling pathways, thereby modulating immune homeostasis in UC rats and effectively alleviating UC.
2.Mechanism of Modified Guizhi Fulingwan in Regulating Mitochondrial Apoptosis Pathway Through PTEN/PI3K/Akt Pathway to Prevent Colorectal Adenoma in Mice
Sujie JIA ; Longhui LIU ; Yifan ZHANG ; Xiaomeng LANG ; Jianping LIU ; Xin KANG ; Chaodi SUN ; Jingyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(19):10-20
ObjectiveTo investigate the effect and mechanism of modified Guizhi Fulingwan in preventing colorectal adenoma (CRA) in mice by regulating mitochondrial apoptosis pathway through the regulation of the phosphatase and tensin homolog (PTEN)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway. MethodSixty SPF-grade male C57BL/6 mice were randomly divided into six groups: Normal group, model group, low, medium, and high dose groups of modified Guizhi Fulingwan (13, 26, 52 g·kg-1·d-1), and positive control aspirin group (0.015 g·kg-1·d-1). A mouse model of CRA was chemically induced using azoxymethane (AOM) and dextran sulfate sodium (DSS). During the modeling process, mice received modified Guizhi Fulingwan or aspirin. Body weight of mice was measured weekly during the treatment. After 9 weeks, the number of adenomas formed was observed. Serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) were determined by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to observe the histopathologic changes in adenoma tissues. The expression of Cyclin D1 and proliferative nuclear antigen (Ki67) was detected by immunohistochemistry (IHC). Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was used to assess the apoptosis in adenoma tissues. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to observe the mRNA and protein expression levels of PTEN, PI3K, Akt, phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-Akt), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cytochrome C (Cyt C), Caspase-9, and caspase-3. ResultCompared with the normal group, the model group showed no significant change in body weight from week 1 to week 2, but a significant decrease from week 3 to week 9 (P<0.05,P<0.01). The colorectal length was significantly shortened, and the colorectal weight increased with visible varying sized tumor-like protrusions on the mucosal surface (P<0.01). Serum levels of TNF-α, IL-6, and IL-1β were elevated (P<0.01). Histopathology showed disordered epithelial gland structure, elongated nuclei with pathological mitosis, and numerous lymphocytic infiltrations in the lamina propria. The positive expression rates of Cyclin D1 and Ki67 were significantly increased (P<0.01), while the apoptosis rate of adenoma cells was significantly decreased (P<0.01). Expression levels of PI3K, Akt, Bcl-2 mRNA and proteins, as well as p-PI3K and p-Akt proteins, were significantly increased (P<0.01), whereas PTEN, Bax, Cyt C, Caspase-9, and Caspase-3 mRNA and protein levels were significantly decreased (P<0.05, P<0.01). Compared with the model group, all drug treatment groups showed an increase in body weight (P<0.01), decreased intestinal weight, increased colorectal length, reduced number of adenomas significantly (P<0.05, P<0.01), and significantly lowered serum levels of TNF-α, IL-6, and IL-1β (P<0.01). Histopathology indicated improved glandular structure and reduced neutrophil infiltration in the mucosal lamina propria. The positive expression rates of Cyclin D1 and Ki67 significantly decreased (P<0.01), while the apoptosis rate of adenoma cells significantly increased (P<0.05, P<0.01). Expression levels of PI3K, Akt, Bcl-2 mRNA and proteins, and p-PI3K and p-Akt proteins were significantly reduced (P<0.05, P<0.01), while PTEN, Bax, Cyt C, Caspase-9, and Caspase-3 mRNA and protein levels significantly increased (P<0.05, P<0.01). The high-dose modified Guizhi Fulingwan group exhibited the most significant intervention effects. ConclusionModified Guizhi Fulingwan may prevent CRA in mice by regulating the PTEN/PI3K/Akt signaling pathway and inducing the mitochondrial apoptosis pathway.
3.Mechanism of Xiezhuo Jiedu Recipe in Preventing Colorectal Adenoma in Mice Through miRNA-34a-5p/PI3K/Akt Signalling Pathway
Sujie JIA ; Chaodi SUN ; Yifan ZHANG ; Xiaomeng LANG ; Jianping LIU ; Xin KANG ; Shijie REN ; Jingyuan LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):156-165
ObjectiveKey microRNAs (miRNAs) of colorectal adenoma (CRA) were identified and analyzed by bioinformatics methods, and differentially expressed genes (DEGs) were screened to construct regulatory relationships. The mechanism of Xiezhuo Jiedu recipe in preventing CRA was speculated and verified by animal experiments. MethodThe miRNAs dataset GSE50194 was obtained from the Gene Expression Omnibus (GEO) database of intestinal mucosal tissue of CRA patients, and the differentially expressed miRNAs were screened by GEO2R and Excel. TargetScan, miRTarbase, and miRDB databases were used to predict the target genes of the differentially expressed miRNAs, and an intersection was obtained. Key DEGs were screened through the STRING database and Cytoscape software, and the TRRUST database was used to predict downstream binding transcription factors (TFs). The mRNA intersection was enriched by gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) in the Metascape database. DIANA TOOLS were applied to perform KEGG enrichment analysis of key miRNAs, and the key signaling pathways were selected for animal experiments. In animal experiments, the CRA mice model was established by using sodium glycan sulfate (DSS) drinking combined with intraperitoneal injection of azomethane oxide (AOM), and Xiezhuo Jiedu recipe and aspirin were given by intragastric administration at the same time. The experiment lasted for nine weeks. The pathological changes in intestinal tissue were observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of miR-34a-5p in adenoma tissue. Protein expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), phosphoryl-PI3K (p-PI3K), phosphoryl-Akt (p-Akt), and B cell lymphoma (Bcl)-2 were detected by Western blot. The expression of Cyclin D1 (CCND1) was detected by immunohistochemistry (IHC). In situ terminal transferase labeling (TUNEL) was used to detect apoptosis of adenoma tissue cells. ResultThe GEO database screened the GSE50194 dataset, and miR-34a-5p was selected as the research object from CRA and normal tissue. A total of 93 DEGs were selected. Among them, GO and KEGG enrichment analyses were closely related to biological processes such as transcriptional regulatory complex, RNA polymerase Ⅱ transcriptional regulatory complex, enzyme-linked receptor protein signaling pathway, and DNA-binding transcriptional activator activity, cancer pathway, PI3K/Akt pathway, etc. miR-34a-5p is mainly enriched in PI3K/Akt, cell cycle, and colorectal cancer pathways. Five key DEGs were screened out through the Matescape database, among which Bcl-2 and CCND1 were the key DEGs of miR-34a-5p. Further screening of the TFs of key DEGs revealed that E2F transcription factor 1 (E2F1) and tumor protein P53 (TP53) were the main TFs of Bcl-2 and CCND1. Animal experiments showed that Xiezhuo Jiedu recipe could effectively up-regulate mRNA level of miR-34a-5p, down-regulate the expression of PI3K, Akt, Bcl-2, p-PI3K, and p-Akt proteins in the intestinal tissue of CRA mice, down-regulate the positive expression rate of CCND1, and increase the apoptosis rate of intestinal epithelial cells. ConclusionIt is speculated that Xiezhuo Jiedu recipe may inhibit the abnormal proliferation and promote the apoptosis of intestinal epithelial cells in CRA mice by regulating the miR-34a-5p/PI3K/Akt signaling pathway, thus playing a role in the prevention of CRA.
4.Evaluation of the efficacy of Internet+cardiac rehabilitation intervention in patients with hypertension and coronary heart disease
Fengxia QU ; Yue XIN ; Jingyuan LI ; Xiao GUO ; Changhong LU
The Journal of Practical Medicine 2024;40(19):2778-2782
Objective To explore the effect of Internet+cardiac rehabilitation on hypertension patients with coronary heart disease.Methods 80 patients with hypertension and coronary heart disease admitted to our hospital from January 2022 to April 2023 were randomly divided into the combined group(n=40)and the conventional group(n=40).Both groups were given routine cardiac rehabilitation interventions.The combined group combined adoptionof the Internet for continuous interventionson the basis of routine cardiac rehabilitation interventions.Compare the oxygen uptake,blood pressure,compliance,and self-management scores between two groups before intervention and three months after discharge.Results The AT Mets,AT VO2,peak VO2,and expected value compliance ratio in the combined group were all higher than those in the conventional group(P<0.05);The systolic and diastolic blood pressure in the combination group were lower than those in the control group,and the proportion of complete compliance was significantly higher than that in the control group(P<0.05);The self-management scores of the joint group were higher than those of the conventional group(P<0.05).Conclusion The continuous interventions of Internet plus cardiac rehabilitation can control the blood pressure of patients with hypertension and coronary heart disease,and patients'compliance behavior is significantly im-proved,andthus improving patients'self-management ability,which can be used for reference and promotion in clinical practice.
5.Establishment of evaluation index system for hospital accreditation data quality based on analytic hier-archy process
Jingyuan XIN ; Yuan YANG ; Wanli MA ; Peifeng LIANG ; Dong ZHU ; Ping SHEN ; Zeqin ZHANG
Modern Hospital 2024;24(8):1182-1185
Objective To establish an index system for assessing the quality of hospital accreditation data and determine the hierarchical weights of indices at different levels,providing a reference for enhancing the governance of such data.Methods A quality assessment system for hospital accreditation data was developed using literature analysis method and Delphi method.The analytic hierarchy process was employed to create a pairwise comparison judgment matrix to determine the weights of indices.Results An evaluation system was constructed with the following levels:"Indicator Screening Quality-Data Reporting Quality-Data Quality Control Quality-Data On-Site Verification Quality."The system includes 4 first-level indicators and 18 second-level indicators.The indicator with the highest weight among the first-level indicators was Data Reporting Quality(0.496 5),followed by Data Quality Control Quality(0.313 2).Among the second-level indicators,Timeliness(0.428 6)and Cooperation(0.428 6)had the highest weights.Conclusion The establishment of the quality evaluation system of hospital accreditation da-ta is a strategy to optimize the control of hospital accreditation data quality and is beneficial for continuous improvement of hospital accreditation data quality.
6.Artificial intelligence-assisted prediction of olfactory disorders in patients with chronic rhinosinusitis.
Jingyuan CHEN ; Wenyi CHEN ; Xin LUO ; Xuekun HUANG ; Yana ZHANG ; Qintai YANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2023;37(11):871-885
Objective:To analyze the influencing factors and perform the prediction of olfactory disorders in patients with chronic rhinosinusitis(CRS) based on artificial intelligence. Methods:The data of 75 patients with CRS who underwent nasal endoscopic surgery from October 2021 to February 2023 in the Department of Otorhinolaryngology Head and Neck Surgery, the Third Affiliated Hospital of Sun Yat-sen University were analyzed retrospectively. There were 53 males and 22 females enrolled in the study, with a median age of 42.0 years old. The CRS intelligent microscope interpretation system was used to calculate the proportion of area glands and blood vessels occupy in the pathological sections of each patient, and the absolute value and proportion of eosinophils, lymphocytes, plasma cells and neutrophils. The patients were grouped according to the results of the Sniffin' Sticks smell test, and the clinical baseline data, differences in nasal mucosal histopathological characteristics, laboratory test indicators and sinus CT were compared between the groups. Determine the independent influencing factors of olfactory disorders and receiver operating characteristic curves(ROC) were used to evaluate the performance of the prediction model. Statistical analysis was performed using SPSS 25.0 software. Results:Among the 75 CRS patients, 25 cases(33.3%) had normal olfaction and 50 cases(66.7%) had olfactory disorders. Multivariate Logistic regression analysis showed that tissue eosinophils percentage(OR=1.032, 95%CI 1.002-1.064, P=0.036), Questionnaire of olfactory disorders-Negative statement(QOD-NS)(OR=1.079, 95%CI 1.004-1.160, P=0.040) and Anterior olfactory cleft score(AOCS)(OR=2.672, 95%CI 1.480-4.827, P=0.001) were independent risk factors for olfactory disorders in CRS patients. Further research found that the area under the ROC curve(AUC) of the combined prediction model established by the tissue eosinophil percentage, QOD-NS and AOCS was 0.836(95%CI 0.748-0.924, P<0.001), which is better than the above single factor prediction model in predicting olfactory disorders in CRS. Conclusion:Based on pathological artificial intelligence, tissue eosinophil percentage, QOD-NS and AOCS are independent risk factors for olfactory disorders in CRS patients, and the combination of the three factors has a good predictive effect on CRS olfactory disorders.
Male
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Female
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Humans
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Adult
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Retrospective Studies
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Artificial Intelligence
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Rhinosinusitis
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Rhinitis/complications*
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Nasal Polyps/complications*
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Sinusitis/complications*
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Olfaction Disorders/etiology*
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Smell
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Chronic Disease
7.Biomonitoring for traditional herbal medicinal products using DNA metabarcoding and single molecule, real-time sequencing.
Tianyi XIN ; Zhichao XU ; Jing JIA ; Christine LEON ; Songnian HU ; Yulin LIN ; Subramanyam RAGUPATHY ; Jingyuan SONG ; Steven G NEWMASTER
Acta Pharmaceutica Sinica B 2018;8(3):488-497
Global concerns have been paid to the potential hazard of traditional herbal medicinal products (THMPs). Substandard and counterfeit THMPs, including traditional Chinese patent medicine, health foods, dietary supplements, etc. are potential threats to public health. Recent marketplace studies using DNA barcoding have determined that the current quality control methods are not sufficient for ensuring the presence of authentic herbal ingredients and detection of contaminants/adulterants. An efficient biomonitoring method for THMPs is of great needed. Herein, metabarcoding and single-molecule, real-time (SMRT) sequencing were used to detect the multiple ingredients in Jiuwei Qianghuo Wan (JWQHW), a classical herbal prescription widely used in China for the last 800 years. Reference experimental mixtures and commercial JWQHW products from the marketplace were used to confirm the method. Successful SMRT sequencing results recovered 5416 and 4342 circular-consensus sequencing (CCS) reads belonging to the ITS2 and regions. The results suggest that with the combination of metabarcoding and SMRT sequencing, it is repeatable, reliable, and sensitive enough to detect species in the THMPs, and the error in SMRT sequencing did not affect the ability to identify multiple prescribed species and several adulterants/contaminants. It has the potential for becoming a valuable tool for the biomonitoring of multi-ingredient THMPs.
8.Value of perioperative multimodal stratified analgesia guided by PPRS-CYMZ 2.0
Lihua PENG ; Su MIN ; Li REN ; Xuechao HAO ; Bo CHENG ; Ping WANG ; Kaihua HE ; Juying JIN ; Jun CAO ; Ke WEI ; Dan LIU ; Yiwei SHEN ; Feng LYU ; Jie DENG ; Xin WANG ; Jun YANG ; Jingyuan CHEN ; Fei XIE
Chinese Journal of Anesthesiology 2017;37(11):1347-1352
Objective To evaluate the value of perioperative multimodal stratified analgesia guided by PPRS-CYMZ 2.0. Methods One hundred and sixteen patients of both sexes, aged 16-85 yr, of A-merican Society of Anesthesiologists physical statusⅠ-Ⅲ, scheduled for elective surgery in our hospital in August 2016, were included in this study and assigned into empirical analgesia group(group E, n=79) and stratified analgesia group(group S, n=73). The risk of postoperative pain was estimated by an expe-rienced associate chief anesthesiologist based on his clinical experience, and the perioperative analgesic protocol was determined in group E. The risk of postoperative pain was assessed using the perioperative pain risk scale PPRS-CYMZ 2.0 by another experienced associate chief anesthesiologist, the risk was stratified according to the scores, and the corresponding stratified analgesic protocol was determined in group S. Vis-ual analog scale scores and parents′satisfaction with analgesia were recorded on postoperative day 30. The requirement for preventive analgesia, total pressing times of patient-controlled analgesia(PCA)pump in 0-6 h, 6-24 h and 24-72 h periods, PCA background infusion dose and consumption of rescue analgesics were recorded. The development of adverse events during postoperative hospital stay and postoperative re-covery were also recorded. Analgesia-related parameters of medical economics were calculated. Results There was no significant difference in postoperative pain risk stratification between group E and group S(P>0.05), and the majority of patients were at moderate risk. Compared with group E, no significant change was found in visual analog scale scores on postoperative day 30, PCA background infusion dose or incidence of postoperative adverse effects(P>0.05), the requirement for preventive analgesia and satisfaction scores were significantly increased in high risk patients, the consumption of rescue analgesics was decreased in moderate risk patients(P<0.05), no significant change was found in the total pressing times of PCA pump in each time period in low risk patients(P>0.05), the total pressing times of PCA pump was significantly decreased, and the direct analgesic cost per patient and total analgesic cost were decreased in moderate and high risk patients, and the first ambulation time and length of postoperative hospital stay were shortened in high risk patients in group S(P<0.05). Conclusion PPRS-CYMZ 2.0 can achieve perioperative multi-modal stratified analgesia and individualized treatment.
9.HCMV-encoded miRNA during latent infection
Xin SONG ; Yanyu ZHANG ; Jiang DENG ; Jun XIAO ; Bo GAO ; Ping MA ; Jingyuan BI ; Xipeng ZHOU ; Jinbo XU
Military Medical Sciences 2016;40(4):311-315
Objective To explore humam cytomegalovirus(HCMV) encoded microRNAs during latent infection in order to help study HCMV virology and latent infection mechanisms.Methods A model of HCMV latent infection via THP-1 cells infected with HCMV was constructed.Deep-sequencing was performed using high-resolution Solexa sequencing platform.The secondary structure of the newly sequenced miRNA was predicted by RNAFold bioinformatics software. Results HCMV encoded various miRNAs during latent infection, including miR-US25-2-3p, miR-US25-2-5p, miR-UL112, miR-US25-1, miR-UL22A and PC-5p-148467 with a predicted length of 25 bp, named hcmv-miR-US33as-5p.Conclusion HCMV can express many types of miRNAs during latent infection.
10.Cloning and analysis of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase genes HsHDR1 and HsHDR2 in Huperzia serrate.
Haizhou LV ; Xin ZHANG ; Baosheng LIAO ; Wanjing LIU ; Liu HE ; Jingyuan SONG ; Chao SUN ; Hongmei LUO ; Shilin CHEN
Acta Pharmaceutica Sinica B 2015;5(6):583-589
We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) gene family from Huperzia serrate. The two transcripts coding HDR, named HsHDR1 and HsHDR2, were discovered in the transcriptome dataset of H. serrate and were cloned by reverse transcription-polymerase chain reaction (RT-PCR). The physicochemical properties, protein domains, protein secondary structure, and 3D structure of the putative HsHDR1 and HsHDR2 proteins were analyzed. The full-length cDNA of the HsHDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids, whereas the HsHDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids. These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (PF02401), but without the transmembrane region and signal peptide. The most abundant expression of HsHDR1 and HsHDR2 was detected in H. serrate roots, followed by the stems and leaves. Our results provide a foundation for exploring the function of HsHDR1 and HsHDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.

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