1.Cuscutae Semen-Lycii Fructus Improves Spermatogenesis in Rat Model of Oligoasthenozoospermia by Inhibiting Oxidative Stress-induced Blood-testis Barrier Damage via Regulating SIRT1/Nrf2 Signaling Pathway
Wen DUAN ; Xiaojing ZHANG ; Wenjie DING ; Jianning JIN ; Guoqing CHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):29-38
ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia (OAS) and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group (n=7) and a modeling group (n=28). The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups: model, Cuscutae Semen-Lycii Fructus granules (3.2 g·kg-1), Cuscutae Semen-Lycii Fructus total flavonoids (0.34 g·kg-1), and L-carnitine (0.38 g·kg-1), and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase (SOD), malondialdehyde (MAD), and glutathione peroxidase (GSH-Px) in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels, respectively, of SIRT1, Nrf2, Occludin, zonula occludens-1 (ZO-1), connexin 43 (CX43), and β-catenin. ResultsCompared with the blank group, the model group showed decreased total sperm count and motility (P<0.05, P<0.01), obvious damage in the testicular tissue and blood-testis barrier structure, reduced score of spermatogenic function (P<0.01), declined levels of GSH-Px and SOD in the testicular tissue (P<0.05), elevated level of MDA, and down-regulated protein levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin (P<0.05, P<0.01) and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, and β-catenin in the testicular tissue (P<0.05, P<0.01). After treatment, the testicular tissue, blood-testis barrier structure, and score of spermatogenic function (P<0.01) were improved in the Cuscutae Semen-Lycii Fructus granules group, Cuscutae Semen-Lycii Fructus total flavonoids group, and L-carnitine group. Compared with the model group, the treatment groups presented lowered levels of GSH-Px and SOD (P<0.05, P<0.01), and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group (P<0.05, P<0.01). ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage, thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.
2.Cuscutae Semen-Lycii Fructus Improves Spermatogenesis in Rat Model of Oligoasthenozoospermia by Inhibiting Oxidative Stress-induced Blood-testis Barrier Damage via Regulating SIRT1/Nrf2 Signaling Pathway
Wen DUAN ; Xiaojing ZHANG ; Wenjie DING ; Jianning JIN ; Guoqing CHU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):29-38
ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia (OAS) and decipher the mechanism based on the silent information regulator 1 (SIRT1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group (n=7) and a modeling group (n=28). The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups: model, Cuscutae Semen-Lycii Fructus granules (3.2 g·kg-1), Cuscutae Semen-Lycii Fructus total flavonoids (0.34 g·kg-1), and L-carnitine (0.38 g·kg-1), and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase (SOD), malondialdehyde (MAD), and glutathione peroxidase (GSH-Px) in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels, respectively, of SIRT1, Nrf2, Occludin, zonula occludens-1 (ZO-1), connexin 43 (CX43), and β-catenin. ResultsCompared with the blank group, the model group showed decreased total sperm count and motility (P<0.05, P<0.01), obvious damage in the testicular tissue and blood-testis barrier structure, reduced score of spermatogenic function (P<0.01), declined levels of GSH-Px and SOD in the testicular tissue (P<0.05), elevated level of MDA, and down-regulated protein levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin (P<0.05, P<0.01) and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, and β-catenin in the testicular tissue (P<0.05, P<0.01). After treatment, the testicular tissue, blood-testis barrier structure, and score of spermatogenic function (P<0.01) were improved in the Cuscutae Semen-Lycii Fructus granules group, Cuscutae Semen-Lycii Fructus total flavonoids group, and L-carnitine group. Compared with the model group, the treatment groups presented lowered levels of GSH-Px and SOD (P<0.05, P<0.01), and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1, Nrf2, ZO-1, CX43, β-catenin, and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group (P<0.05, P<0.01). ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage, thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.
3.Traditional Chinese Medicine Treats Colorectal Cancer by Regulating PI3K/Akt/mTOR Signaling Pathway: A Review
Yingying SUN ; Pan ZHENG ; Jin DING
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):271-281
Colorectal cancer (CRC) is a prevalent malignant tumor of the digestive tract, with a high incidence and high mortality. The majority of patients are diagnosed at the middle or advanced stage, which severely influences and threatens their physical health. Current treatment modalities such as surgery, radiotherapy, and chemotherapy often encounter challenges including metastasis, recurrence, and drug resistance. The phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway serves as a classical regulator that regulates physiological processes such as cell cycle, autophagy, apoptosis, and proliferation. Overexpression of this pathway is observed in various tumors. In the context of CRC, the activation of this pathway can facilitate the proliferation, invasion, and migration, inhibit the autophagy and apoptosis, promote the epithelial-mesenchymal transition of CRC cells, enhance angiogenesis within the tumor, and contribute to chemotherapy resistance and radiation resistance in CRC. Traditional Chinese medicine (TCM) treatment can exert an anti-CRC effect by inhibiting this pathway, thereby improving clinical efficacy and safety. This article retrieves relevant research literature published domestically and internationally regarding the regulation of the PI3K/Akt/mTOR signaling pathway by TCM in the treatment of CRC and conducts detailed classification and summary. The active components of TCM include glycosides, flavonoids, alkaloids, terpenoids, polyphenols, and naphthoquinones. The volatile oils and extracts of TCM include Angelicae Sinensis Radix volatile oil, Astragali Radix polysaccharides, Caryophylli Flos extract, Forsythiae Fructus extract, Curcumae Longae Rhizoma extract, and Celastrus orbiculatus extract. The compound formulas of TCM include Banxia Xiexin decoction, Jianpi Qingre Huoxue formula, and Chanling Plaster. Through summary and analysis, it is discovered that the abovementioned TCM can produce effects such as blocking the cell cycle, inducing autophagy and apoptosis, inhibiting angiogenesis, suppressing proliferation and migration, and reversing chemotherapy resistance and radiotherapy resistance by inhibiting the PI3K/Akt/mTOR pathway in CRC cells. TCM holds promise in the research and application of targeting the PI3K/Akt/mTOR signaling pathway for CRC treatment. The summary and conclusion of this article aim to provide references for subsequent research and the development of new drugs.
4.Traditional Chinese Medicine Treats Colorectal Cancer by Regulating PI3K/Akt/mTOR Signaling Pathway: A Review
Yingying SUN ; Pan ZHENG ; Jin DING
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):271-281
Colorectal cancer (CRC) is a prevalent malignant tumor of the digestive tract, with a high incidence and high mortality. The majority of patients are diagnosed at the middle or advanced stage, which severely influences and threatens their physical health. Current treatment modalities such as surgery, radiotherapy, and chemotherapy often encounter challenges including metastasis, recurrence, and drug resistance. The phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway serves as a classical regulator that regulates physiological processes such as cell cycle, autophagy, apoptosis, and proliferation. Overexpression of this pathway is observed in various tumors. In the context of CRC, the activation of this pathway can facilitate the proliferation, invasion, and migration, inhibit the autophagy and apoptosis, promote the epithelial-mesenchymal transition of CRC cells, enhance angiogenesis within the tumor, and contribute to chemotherapy resistance and radiation resistance in CRC. Traditional Chinese medicine (TCM) treatment can exert an anti-CRC effect by inhibiting this pathway, thereby improving clinical efficacy and safety. This article retrieves relevant research literature published domestically and internationally regarding the regulation of the PI3K/Akt/mTOR signaling pathway by TCM in the treatment of CRC and conducts detailed classification and summary. The active components of TCM include glycosides, flavonoids, alkaloids, terpenoids, polyphenols, and naphthoquinones. The volatile oils and extracts of TCM include Angelicae Sinensis Radix volatile oil, Astragali Radix polysaccharides, Caryophylli Flos extract, Forsythiae Fructus extract, Curcumae Longae Rhizoma extract, and Celastrus orbiculatus extract. The compound formulas of TCM include Banxia Xiexin decoction, Jianpi Qingre Huoxue formula, and Chanling Plaster. Through summary and analysis, it is discovered that the abovementioned TCM can produce effects such as blocking the cell cycle, inducing autophagy and apoptosis, inhibiting angiogenesis, suppressing proliferation and migration, and reversing chemotherapy resistance and radiotherapy resistance by inhibiting the PI3K/Akt/mTOR pathway in CRC cells. TCM holds promise in the research and application of targeting the PI3K/Akt/mTOR signaling pathway for CRC treatment. The summary and conclusion of this article aim to provide references for subsequent research and the development of new drugs.
5.Development and research of an AI-assisted decision-making platform in treatment of insomnia with acupuncture of Tongdu Yangxin acupoint prescription.
Chi WANG ; Chengyong LIU ; Xiaoqiu WANG ; Enqi LIU ; Juguang SUN ; Jin LU ; Min DING ; Wenzhong WU
Chinese Acupuncture & Moxibustion 2025;45(7):881-888
OBJECTIVE:
To construct and validate a predictive model for the therapeutic effect of acupuncture at Tongdu Yangxin prescription (acupoint prescription for promoting the circulation of the governor vessel and nourishing the heart) on insomnia, so as to develop an open-access interactive artificial intelligence (AI)-assisted decision-making platform.
METHODS:
Clinical data of 139 insomnia patients treated with Tongdu Yangxin acupuncture therapy were included. All the patients had received acupuncture at Baihui (GV20), Yintang (GV24+), bilateral Shenmen (HT7), and bilateral Sanyinjiao (SP6); and electric stimulation was attached to Baihui (GV20) and Yintang (GV24+), using a continuous wave and a frequency of 2 Hz. The treatment was delivered once every other day, 3 treatments a week, and for 2 consecutive weeks. Patients with Pittsburgh sleep quality index (PSQI) score reduction rate <50% were classified as the "no response group", and those with ≥50% were as the "response group". Outliers were addressed using the 1.5×IQR rule, and missing values were imputed via predictive mean matching. Key features were selected by intersecting the feature importance results from eXtreme Gradient Boosting (XGBoost) and random forest algorithms. After balancing class distribution using the Synthetic Minority Over-sampling Technique (SMOTE), 20% of the data was reserved as a validation set. The remained data underwent the stratified sampling iterations to generate 200 pairs of 3∶1 training-test sets, which was employed for training and internal validation of 8 machine learning algorithms. The optimal algorithm and data partitioning strategy were selected to construct the final model, followed by external validation. The best-performing model was deployed online via Streamlit to create an interactive AI platform.
RESULTS:
Key predictive features for model construction included insomnia duration, the total PSQI score, PSQI sleep efficiency subscore, the proportion of N1 and N2 sleep stages in total sleep duration, and the maximum pulse rate during sleep. The CatBoost-based model achieved an AUC of 0.92, the average precision of 0.77, and accuracy, average recall, and average F1-score of 0.75 on the test set. On the validation set, it attained an AUC of 0.84, with accuracy, average precision, average recall, and average F1-score all at 0.72, demonstrating robust predictive performance. An interactive AI platform was subsequently developed (https://tdyx-catboost.streamlit.app/).
CONCLUSION
This study successfully establishes and validates a CatBoost-based efficacy prediction model for Tongdu Yangxin acupuncture therapy in treatment of insomnia. The developed AI platform provides data-driven decision support for acupuncture-based insomnia management.
Humans
;
Sleep Initiation and Maintenance Disorders/physiopathology*
;
Acupuncture Therapy
;
Male
;
Acupuncture Points
;
Female
;
Middle Aged
;
Adult
;
Artificial Intelligence
;
Aged
;
Young Adult
6.De novo patients with high-volume metastatic hormone-sensitive prostate cancer can benefit from the addition of docetaxel to triplet therapy: Network-analysis and systematic review.
Hanxu GUO ; Chengqi JIN ; Li DING ; Jun XIE ; Jing XU ; Ruiliang WANG ; Hong WANG ; Changcheng GUO ; Jiansheng ZHANG ; Bo PENG ; Xudong YAO ; Jing YUAN ; Bin YANG
Chinese Medical Journal 2025;138(2):231-233
7.Analysis of effectiveness of Holosight robot navigation-assisted percutaneous cannulated screw fixation in treatment of femoral neck fractures.
Weizhen XU ; Zhenqi DING ; Hui LIU ; Jinhui ZHANG ; Yuanfei XIONG ; Jin WU
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(6):673-679
OBJECTIVE:
To investigate the effectiveness of Holosight robotic navigation-assisted percutaneous cannulated screw fixation for femoral neck fractures.
METHODS:
A retrospective analysis was conducted on 65 patients with femoral neck fractures treated with cannulated screw fixation between January 2022 and February 2024. Among them, 31 patients underwent robotic navigation-assisted screw placement (navigation group), while 34 underwent conventional freehand percutaneous screw fixation (freehand group). Baseline characteristics, including age, gender, fracture side, injury mechanism, Garden classification, Pauwels classification, and time from injury to operation, showed no significant differences between the two groups ( P>0.05). The operation time, intraoperative blood loss, fluoroscopy frequency, fracture healing time, and complications were recorded and compared, and hip function was evaluated by Harris score at last follow-up. Postoperative anteroposterior and lateral hip X-ray films were taken to assess screw distribution accuracy, including deviation from the femoral neck axis, inter-screw parallelism, and distance from screws to the femoral neck cortex.
RESULTS:
No significant difference was observed in operation time between the two groups ( P>0.05). However, the navigation group demonstrated superior outcomes in intraoperative blood loss, fluoroscopy frequency, deviation from the femoral neck axis, inter-screw parallelism, and distance from screws to the femoral neck cortex ( P<0.05). No incision infections or deep vein thrombosis occurred. All patients were followed up 12-18 months (mean, 16 months). In the freehand group, 1 case suffered from cannulated screw dislodgement and nonunion secondary to osteonecrosis of femoral head at 1 year after operation, 1 case suffered from screw penetration secondary to osteonecrosis of femoral head at 5 months after operation; and 1 case suffered from nonunion secondary to osteonecrosis of femoral head at 6 months after operation in the navigation group. All the 3 patients underwent internal fixators removal and total hip arthroplasty. There was no significant difference in the incidence of complications between the two groups ( P>0.05). The fracture healing time and hip Harris score at last follow-up in the navigation group were significantly better than those in the freehand group ( P<0.05).
CONCLUSION
Compared to freehand percutaneous screw fixation, Holosight robotic navigation-assisted cannulated screw fixation for femoral neck fractures achieves higher precision, reduced intraoperative radiation exposure, smaller incisions, and superior postoperative hip function recovery.
Humans
;
Femoral Neck Fractures/diagnostic imaging*
;
Bone Screws
;
Fracture Fixation, Internal/instrumentation*
;
Male
;
Female
;
Retrospective Studies
;
Robotic Surgical Procedures/methods*
;
Middle Aged
;
Aged
;
Adult
;
Treatment Outcome
;
Operative Time
;
Fracture Healing
;
Surgery, Computer-Assisted/methods*
;
Fluoroscopy
8.Comparative study on lengthened proximal femoral nail antirotation and InterTan in the treatment of subtrochanteric femur fractures in the elderly.
Zhen-Shan WANG ; Ming-Cong DING ; Jin JIANG ; Jing-Sheng LIU ; Tong-Qun YANG
China Journal of Orthopaedics and Traumatology 2025;38(7):662-667
OBJECTIVE:
To investigate the clinical efficacy of lengthened proximal femoral nail antirotation (PFNA) and InterTan in the treatment of subtrochanteric femur fractures in the elderly.
METHODS:
A retrospective analysis was performed on the data of 65 elderly patients diagnosed with subtrochanteric femoral fractures who met the inclusion criteria between October 2016 and January 2022. The enrolled patients were categorized into two groups according to the type of internal fixation used: the lengthened proximal femoral nail antirotation(PFNA) group and the InterTan group. There were 32 patients in the PFNA group, comprising 20 males and 12 females, with ages ranging from 60 to 85 years old with an average of (69.3±6.7 ) years old. Among these patients, 15 patients resulted from traffic accidents and 17 patients were caused by falls. According to the Seinsheimer classification system, there were 11 patients as type Ⅱ, 14 patients as type Ⅲ, 6 patients as type Ⅳ, and 1 patient as type Ⅴ. The InterTan group comprised 33 patients, including 20 males and 13 females, aged from 60 to 85 years old with an average of (69.8±7.8 ) years old. Of these, 15 patients resulted from traffic accidents, while 18 patients were caused by falls. According to the Seinsheimer classification system, 10 patients as type Ⅱ, 15 patients as type Ⅲ, 7 patients as type Ⅳ, and 1 patient as type Ⅴ. The intraoperative blood loss, operative duration, and fracture healing time were recorded and compared between two groups. The quality of fracture reduction was assessed using Chang's criteria. Additionally, the Harris hip score was utilized to evaluate hip function in both groups at 3 months postoperatively and at the final follow-up.
RESULTS:
All patients were followed up for a period ranging from 10 to 20 months with an average of (14.6±4.5) months. No statistically significant differences were observed between two groups in terms of operation time, intraoperative blood loss, quality of fracture reduction, or reduction methods (P>0.05). Three months after the surgery, the Harris hip score in the InterTan group was 80.0(78.0, 83.5) points, which was significantly higher than that in the PFNA group, which recorded a score of 77.5(75.0, 81.8) points. This difference was statistically significant (P<0.05). At the final follow-up, the Harris hip score in the InterTan group was 80.0(76.5, 87.0), while that in the PFNA group was 78.0(74.3, 82.8). No statistically significant difference was observed between two groups (P>0.05).
CONCLUSION
The use of lengthened PFNA and InterTan in the treatment of elderly subtrochanteric femur fractures can both achieve good treatment results, with the advantages of simple operation, firm fixation, and low failure rate of internal fixation. The lengthened InterTan can achieve better hip function than PFNA.
Humans
;
Male
;
Female
;
Aged
;
Aged, 80 and over
;
Bone Nails
;
Retrospective Studies
;
Hip Fractures/surgery*
;
Middle Aged
;
Fracture Fixation, Intramedullary/instrumentation*
;
Fracture Fixation, Internal/methods*
;
Femoral Fractures/surgery*
9.Analysis of ABO System Hemolytic Disease of the Newborn in 283 Cases at Yunnan Province.
Jin-Yu ZHOU ; Ru SHEN ; Han-Xin WU ; Ju-Ding GUO ; Hong-Mei LIU ; Li-Li SHU ; Yu ZHU ; Jing-Yue SUN ; Jun CHANG
Journal of Experimental Hematology 2025;33(3):881-885
OBJECTIVE:
To analyze the laboratory detection results of hemolytic disease of the fetus and newborn(HDFN).
METHODS:
Related test results of 283 newborns and their mothers' blood samples from Kunming Maternal and Child Health Hospital from August 2023 to May 2024 were collected, including mother and child ABO blood group, RhD blood group, as well as 3 tests of HDFN, total bilirubin (TBil) and indirect bilirubin (IBil).
RESULTS:
283 were ABO incompatibility, among which 187 were HDFN positive, with a positive rate of 66.08%; the positive rate of HDFN in neonates with antigen-A incompatibility was 74.12%(126/170), the positive rate of HDFN in neonates with antigen-B incompatibility was 53.57%(60/112), which was the highest in neonates with O/A incompatibility [75.45%(126/167)], followed by O/B incompatibility[54.55%(60/110)]. Group by age, the positive rates of HDFN in the ≤1 d group, 2 d group, 3 d group, 4 d group, 5 d group and ≥6 d group were 76.03%(111/146), 67.86%(38/56), 57.14%(24/42), 38.46%(5/13), 46.15%(6/13) and 23.08%(3/13), respectively. With the increase of age, the positive rates of HDFN gradually decreased, there was a statistically significant difference between the ≤3 day age group and >3 day age group ( P <0.05). There was no statistically significant difference in TBil and IBil levels between the "direct antibody+indirect antibody+release+" group and the HDFN negative group in newborns. HDFN infants exhibited a rapid increase in bilirubin levels within the first day after birth, with significantly higher TBil and IBil values compared to Non ABO-HDFN infants in the ≤1 day group ( P <0.01). However, the difference of bilirubin levels between the two groups gradually narrowed from 2-6 days after birth, and the difference was not statistically significant (P >0.05). The peak value of TBil and IBil occurred on the 4th day after birth in HDFN infants.
CONCLUSION
ABO-HDFN is most commonly seen in newborns whose mothers are type-O, and the positive rate was the highest in newborns with O/A incompatibility. The detection rate of HDFN is affected by the age of the newborns, and the two were correlated inversely. ABO-HDFN group developed more rapidly with a higher peak. Therefore, HDFN tests should be carried out as soon as possible for mothers and newborns with incompatible blood types, and appropriate treatment should be provided to prevent complications.
Humans
;
Infant, Newborn
;
ABO Blood-Group System
;
Erythroblastosis, Fetal/epidemiology*
;
Female
;
China/epidemiology*
;
Blood Group Incompatibility
;
Male
;
Bilirubin/blood*
10.A chromosome-level Dendrobium moniliforme genome assembly reveals the regulatory mechanisms of flavonoid and carotenoid biosynthesis pathways.
Jiapeng YANG ; Qiqian XUE ; Chao LI ; Yingying JIN ; Qingyun XUE ; Wei LIU ; Zhitao NIU ; Xiaoyu DING
Acta Pharmaceutica Sinica B 2025;15(4):2253-2272
Dendrobium moniliforme (D. moniliforme) is a traditional medicinal herb widely cultivated in Asia. Flavonoids, one of the largest groups of secondary metabolites in plants, are significant medicinal components in Dendrobium species. Several subgroups of R2R3-MYB proteins have been validated to directly regulate flavonoid biosynthesis. Using PacBio sequencing technology, we assembled a high-quality chromosome-level D. moniliforme genome with a total length of 1.20 Gb and a contig N50 of 3.97 Mb. The BUSCO assessment of genome annotation was 91.4%. By integrating the genome and transcriptome, we identified biosynthesis pathway enzyme genes related to flavonoids, polysaccharides, carotenoids, and alkaloids. A total of 90 R2R3-MYBs were identified in D. moniliforme and classified into 21 subgroups. Studies on the functions of R2R3-MYB transcription factors revealed that R2R3-MYB in SG6 can up-regulate flavonoid biosynthesis. Various validation experiments, including subcellular localization, transient overexpression, UPLC-MS/MS, HPLC, yeast one-hybrid, and dual-luciferase assays, demonstrated that DMYB69 directly up-regulates the expression of enzyme genes involved in flavonoid biosynthesis, increasing the content of flavonoids such as anthocyanin, flavone, and flavonol. Additionally, DMYB44 was shown to directly up-regulate the expression of carotenoid biosynthesis enzyme genes, thereby increasing carotenoid content. This study provides an essential genome resource and theoretical basis for molecular breeding research in D. moniliforme.

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