1.In Vitro and in vivo Component Analysis of Total Phenolic Acids from Gei Herba and Its Effect on Promoting Acute Wound Healing and Inhibiting Scar Formation
Xixian KONG ; Guanghuan TIAN ; Tong WU ; Shaowei HU ; Jie ZHAO ; Fuzhu PAN ; Jingtong LIU ; Yong DENG ; Yi OUYANG ; Hongwei WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):156-167
ObjectiveBased on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), to identify the in vivo and in vitro chemical components of total phenolic acids in Gei Herba(TPAGH), and to clarify the pharmacological effects and potential mechanisms of the effective part in promoting acute wound healing and inhibiting scar formation. MethodsUPLC-Q-Orbitrap-MS was used to identify the chemical components of TPAGH and ingredients absorbed in vivo after topical administration. A total of 120 ICR mice were randomly divided into the model group, recombinant human epidermal growth factor(rhEGF) group(4 mg·kg-1), and low, medium, and high dose groups of TPAGH(3.5, 7, 14 mg·kg-1), with 24 mice in each group. A full-thickness skin excision model was constructed, and each administration group was coated with the drug at the wound site, and the model group was treated with an equal volume of normal saline, the treatment was continued for 30 days, during which 8 mice from each group were sacrificed on days 6, 12, and 30. The healing of the wounds in the mice was observed, and histopathological changes in the skin tissues were dynamically observed by hematoxylin-eosin(HE), Masson, and Sirius red staining, and enzyme-linked immunosorbent assay(ELISA) was used to dynamically measure the contents of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), vascular endothelial growth factor A(VEGFA), matrix metalloproteinase(MMP)-3 and MMP-9 in skin tissues. Network pharmacology was used to predict the targets related to the promotion of acute wound healing and the inhibition of scar formation by TPAGH, and molecular docking of key components and targets was performed. Gene Ontology(GO) biological process analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were carried out for the related targets, so as to construct a network diagram of herbal material-compound-target-pathway-pharmacological effect-disease for further exploring its potential mechanisms. ResultsA total of 146 compounds were identified in TPAGH, including 28 phenylpropanoids, 31 tannins, 23 triterpenes, 49 flavonoids, and 15 others, and 16 prototype components were found in the serum of mice. Pharmacodynamic results showed that, compared with the model group, the TPAGH groups showed a significant increase in relative wound healing rate and relative scar inhibition rate(P<0.05), and the number of new capillaries, number of fibroblasts, number of new skin appendages, epidermal regeneration rate, collagen deposition ratio, and Ⅲ/Ⅰ collagen ratio in the tissue were significantly improved(P<0.05, 0.01), the levels of IL-6, TNF-α, MMP-3 and MMP-9 in the skin tissues were reduced to different degrees, while the level of VEGFA was increased. Network pharmacology analysis screened 10 core targets, including tumor protein 53(TP53), sarcoma receptor coactivator(SRC), protein kinase B(Akt)1, signal transducer and activator of transcription 3(STAT3), epidermal growth factor receptor(EGFR) and so on, participating in 75 signaling pathways such as advanced glycation end-products(AGE)-receptor for AGE(AGE/RAGE) signaling pathway, phosphatidylinositol 3-kinase(PI3K)/Akt signaling pathway, mitogen-activated protein kinase(MAPK) signaling pathway. Molecular docking confirmed that the key components genistein, geraniin, and casuariin had good binding ability to TP53, SRC, Akt1, STAT3 and EGFR. ConclusionThis study comprehensively reflects the chemical composition of TPAGH and the absorbed components after topical administration through UPLC-Q-Orbitrap-MS. TPAGH significantly regulates key indicators of skin healing and tissue reconstruction, thereby clarifying its role in promoting acute wound healing and inhibiting scar formation. By combining in vitro and in vivo component identification with network pharmacology, the study explores how key components may bind to targets such as TP53, Akt1 and EGFR, exerting therapeutic effects through related pathways such as immune inflammation and vascular regeneration.
2.Investigation on the basic situation of pre-analytical quality management in blood station laboratories in North China
Jing SUN ; Hongwei GE ; Zhengmin LIU ; Qianqian QIN ; Wei HAN ; Tong PAN ; Dongli JIAO ; Xiaolan DONG ; Rui WANG
Chinese Journal of Blood Transfusion 2025;38(11):1514-1520
Objective: To investigate the basic situation of pre-analytical quality management in blood station laboratories in North China, and to provide baseline data for promoting the homogenization and standardization of these pre-analytical processes in each blood station laboratory. Methods: A cross-sectional status survey was designed based on the quality management regulations of blood stations, ISO15189 standards and relevant quality management requirements. This survey covering various aspects including laboratory general situation, sample collection and temporary storage, transportation, reception, and quality continuous improvement situations. Data analysis was performed on the survey results of each laboratory. Results: All the 38 blood station laboratories in North China had established a pre-analytical quality management system framework and implemented basic pre-analytical quality control activities; however, there were differences in implementation. 1) Among the 12 basic quality items, 3 items were monitored by all the investigated laboratories (100%), 6 items were monitored by the vast majority of laboratories (about 90%), and 3 items were monitored by a portion of laboratories (about 60%). There were no significant differences in the monitoring index among the three regions and among different types of laboratories (P>0.05). 2) Among the total of 26 items in the three key processes before testing (sample collection and storage, transportation, reception and processing), 12 items were monitored by all laboratories (100%), 11 items were monitored by the vast majority of laboratories (about 90%), and 3 items were monitored by a portion of laboratories (about 75%). There were no significant differences in monitoring index among different regions and types of laboratories (P>0.05). Conclusion: This survey provides a reference and basis for the gap analysis of the pre-analytical process quality management in 38 blood station laboratories across North China. It facilitates laboratories in identifying pre-analytical quality problems, resolving problems, preventing errors, and ensuring that the quality of blood samples before testing meets the established requirements. It lays a foundation for the homogenization of pre-analytical quality management in regional blood stations.
3.Analysis of unqualified pre-analytical samples in blood station laboratories in North China
Zhengmin LIU ; Hongwei GE ; Qianqian QIN ; Wei HAN ; Tong PAN ; Dongli JIAO ; Xiaolan DONG ; Rui WANG
Chinese Journal of Blood Transfusion 2025;38(11):1521-1528
Objective: To determine the frequency and main reasons of unqualified samples by analyzing the quality of pre-analytical samples in blood stations in North China, thereby providing a reference and basis for gap analysis in the implementation of pre-analytical process quality management for participating laboratories and ensuring that only high-standard and high-quality blood samples proceed to testing. Methods: Data on the quality of pre-analytical samples from blood station laboratories in North China was collected via questionnaire. Statistical analysis were performed on: 1) the basic information of samples quality monitoring in the laboratories; 2) the distribution of the overall pre-analytical unqualified rate of samples and the pre-analytical unqualified rate of samples in each laboratory; 3) the distribution of reasons for sample disqualification. Results: 1) The overall pre-analytical unqualified rate of samples in blood station laboratories in North China was 4.55, with a total sigma level of 5.39σ. The 25th, 50th and 75th percentiles (P25, P50, P75) for the total unqualified rate were 0.00, 1.10 and 5.96, respectively. The corresponding percentiles for the Sigma level were 5.34σ, 5.71σ, and 6.00σ, respectively. The pre-analytical unqualified rate of serological and nucleic acid samples (4.89 vs 4.22) showed a significant difference (χ
=9.575, P<0.05). 2) The average unqualified rate of samples in region A, B and C was 1.71, 9.50 and 12.64 (χ
=1 590.721, P<0.05), and the sigma level was 5.66σ, 5.21σ and 5.16σ, respectively. 3) The main reasons for unqualified serological samples were chylous blood (72.65%), hemolysis (17.39%), abnormal hematocrit (5.80%), and insufficient volume (3.50%). The main reasons for the unqualified nucleic acid samples were chylous blood (78.26%), hemolysis (8.84%), failure to centrifuge as required (5.01%), abnormal hematocrit (4.66%), and insufficient volume (1.92%). Conclusion: In North China, the quality indicators for the pre-analytical processes in blood station laboratories are generally well-managed. Laboratories in region A outperformed the national average in pre-analytical specimen quality control. However, participating laboratories exhibit gaps in implementing pre-analytical quality management. Through effective analysis of pre-analytical process quality metrics and inter-laboratory comparisons, laboratories can identify discrepancies and address shortcomings. By establishing clear quality objectives, they can achieve continuous improvement and ensure the validity of test results.
4.Investigation on the management of hemolytic and lipemic samples in the preanalytical phase in blood station laboratories in North China
Jing SUN ; Hongwei GE ; Zhengmin LIU ; Qianqian QIN ; Wei HAN ; Tong PAN ; Dongli JIAO ; Xiaolan DONG ; Rui WANG
Chinese Journal of Blood Transfusion 2025;38(11):1529-1534
Objective: To investigate the assessment criteria and subsequent handling practices of hemolytic and lipemic blood samples before testing in blood screening laboratories in North China, and to provide data to support the standardization of their management in blood station laboratories. Methods: Data on the preanalytical management of hemolytic and lipemic samples from 38 laboratories were collected. The details of management on the criteria and verificatioon for assessment, the assessment methods, and subsequent handling procedures of hemolytic and lipemic samples in blood station laboratories were analyzed. Results: 1) All 38 blood station laboratories monitored serological and nucleic acid samples for hemolysis and lipemia in pre-analytical phase. 2) The criteria and methods for assessing hemolytic and lipemic samples varied among the laboratories of the 38 blood stations. 15 laboratories (39.47%) followed manufacturer's instructions, 9 laboratories (23.68%) formulated their own criteria, and 14 laboratories (36.84%) referred to the criteria of other laboratories. 16 laboratories (42.11%) verified the criteria for assessing hemolytic and lipemic samples, with significant variations in verification rate across laboratories from different regions (P<0.05). For the assessment methods, visual inspection was used by 28 laboratories (73.68%) for hemolytic samples and by 27 laboratories (71.05%) for lipemic samples; the colorimetric card method was used by 10 laboratories (26.32%) for assessing both hemolytic and lipemic samples; the instrumental method was used by 1 laboratory (2.63%) for assessing lipemic samples.3) The handling procedures for hemolytic and lipemic samples varied significantly and followed a gradient distribution pattern among 38 laboratories (including accepting samples for testing, accepting samples for concession testing, re-collecting samples, and rejecting samples and halting testing). With increasing severity of hemolysis and lipemia, more laboratories halted testing, and relatively fewer laboratories accepted samples for normal testing. 5 laboratories (13.16%) applied different handling procedures on serological and nucleic acid samples. Conclusion: This survey provides a reference and basis for analyzing gaps in the management of hemolytic and lipemic samples during the preanalyical phase in blood station laboratories in North China. It enables laboratories to identify the problems and deficiencies in the management of hemolytic and lipemic samples, to ensure preanalytical samples quality meets the established requirements, and to lay a foundation for promoting the homogenization and standardization of the regional sample quality management mode.
6.Photodynamic enhancement of PROTAC prodrug activation in hypoxic tumors.
Zhongliang FU ; Chunrong YANG ; Yuchen YANG ; Meichen PAN ; Hongwei HOU ; Jinghong LI
Acta Pharmaceutica Sinica B 2025;15(9):4945-4960
Proteolysis-targeting chimeras (PROTACs) have emerged as a promising therapeutic strategy for targeted protein degradation. However, the clinical application of PROTACs may be hindered by off-target toxicity resulting from non-tissue-specific protein degradation and ingenious prodrug strategies may open new avenues to addressing this concern. Herein, we propose a light-induced positive feedback strategy to use photodynamic therapy (PDT) to improve the activation efficiency of PROTAC prodrugs, monitor PROTAC release, and combine PROTAC to induce tumor cell apoptosis. In the hypoxic tumor microenvironment, the azo bond in AZO-PRO selectively cleaves, triggering the release of the potent protein degrader PRO and the multifunctional photosensitizer. Once activated, the fluoresce signal of the photosensitizer dramatically recovers, allowing monitoring of prodrug activation. Additionally, upon irradicating the tumor site using near-infrared (NIR) laser, PDT exacerbates tumor hypoxia, further promoting AZO-PRO activation. Our work introduces a novel approach to efficiently track and activate PROTAC prodrugs, enhance their antitumor efficacy, and mitigate off-target systemic toxicity.
7.Stewed Polygoni Multiflori Radix Treats Androgenic Alopecia in Mice by Activating Wnt/β-catenin Signaling Pathway
Fuzhu PAN ; Mingxia CHEN ; Bin YI ; Yanhua XUE ; Qiuping YU ; Fayun WU ; Enhui JI ; Hongwei WU ; Jing XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):246-253
ObjectiveTo evaluate the therapeutic effect of stewed Polygoni Multiflori Radix on androgenic alopecia (AGA) and study the treatment mechanism. MethodNinety-nine SPF-grade male C57BL/6J mice were randomized into control, model, positive drug (finasteride, 0.65 mg·kg-1), low (0.78 g·kg-1), medium (1.56 g·kg-1), and high (3.12 g·kg-1)-dose stewed Polygoni Multiflori Radix, and Polygoni Multiflori Radix Praeparata groups by the random number table method. The mouse model of AGA was constructed by subcutaneous multi-point injection of testosterone propionate diluent for 60 days, and the mice were administrated with corresponding drugs by gavage from day 11. The therapeutic effects of stewed Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata on AGA were evaluated by newly hair area, hair length, hair weight in the hair removal area, and hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was employed to determine the levels of testosterone (T), dihydrotestosterone (DHT), and 5α-reductase (5-AR) in the skin tissue of mice. Western blot was employed to determine the expression levels of key proteins in the Wnt/β-catenin signaling pathway. ResultCompared with the control group, the model group (after 60 days of modeling) showed reductions in the newly hair area, hair length and weight in the back hair removal area, and ratio of hair follicles containing melanin to total hair follicles (P<0.05, P<0.01), elevated levels of T, DHT, and 5-AR, up-regulated expression level of glycogen synthase kinase-3β (GSK-3β) (P<0.05, P<0.01), and down-regulated expression levels of β-catenin, phospho-glycogen synthase kinase-3β (p-GSK-3β), and p-GSK-3β/GSK-3β (P<0.05, P<0.01) in the skin tissue. Compared with the model group, the positive drug, low-, medium-, and high-dose stewed Polygoni Multiflori Radix, and low-, medium-, and high-dose Polygoni Multiflori Radix Praeparata improved the newly hair area and hair length of mice (P<0.01), and stewed Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata at low and medium doses improved the weight of newly formed hair in mice (P<0.05, P<0.01). The positive drug, low-, medium-, and high-dose stewed Polygoni Multiflori Radix, and low- and high-dose Polygoni Multiflori Radix Praeparata increased the ratio of hair follicles containing melanin to total hair follicles in the skin tissue (P<0.05, P<0.01). Compared with Polygoni Multiflori Radix Praeparata at the same doses, the medium and high doses of stewed Polygoni Multiflori Radix increased the ratio of melanin-containing hair follicles to total hair follicles (P<0.05). Compared with the model group, stewed Polygoni Multiflori Radix lowered the levels of T and DHT, down-regulated the expression level of GSK-3β (P<0.01), and up-regulated the expression levels of β-catenin, p-GSK-3β, and p-GSK-3β/GSK-3β (P<0.05, P<0.01) in the skin tissue of the mice. ConclusionStewed Polygoni Multiflori Radix can ameliorate androgenic alopecia in mice by reducing the androgen level and promoting Wnt/β-catenin signaling.
8.Efficacy of Tanzhuo Decoction in the treatment of early diabetic nephropathy in patients with type 2 diabetes mellitus and its effect on cystatin C, C-reactive protein, urinary albumin excretion rate, and creatinine clearance rate
Hongwei YU ; Zhongliang PAN ; Shaorong FAN ; Guoqiang XU
Chinese Journal of Primary Medicine and Pharmacy 2024;31(2):218-222
Objective:To investigate the efficacy of Tanzhuo Decoction in the treatment of early diabetic nephropathy in patients with type 2 diabetes mellitus and its effect on cystatin C (Cys-C), C-reactive protein (CRP), urinary albumin excretion rate (UAER), and creatinine clearance rate (CCr). Methods:Eighty patients with type 2 diabetes mellitus complicated by early diabetic nephropathy who received treatment at Maanshan Hospital of Traditional Chinese Medicine from 2019 to 2021 were included in this randomized controlled study. They were divided into a control group ( n = 40) and a treatment group ( n = 40) using the random number table method. Patients in the control group received conventional therapy including blood glucose and blood pressure control, while those in the treatment group received Tangzhuo Decoction in addition to the same treatment as that given to the control group. Both groups of patients were treated for 30 days. The clinical efficacy as well as pre- and post-treatment Cys-C, CRP, UAER, and CCr were compared between the two groups. Results:The total response rate in the treatment group was 92.5% (37/40), which was significantly higher than 75.0% (30/40) in the control group ( χ2 = 4.50, P < 0.05). After treatment, Cys-C, CRP, and UAER in the treatment group were (2.04 ± 0.08) mg/L, (3.97 ± 1.71) mg/L, and (91.18 ± 18.68) μg/min, respectively, which were significantly decreased compared with those before treatment ( t = 12.14, 5.59, 4.73, all P < 0.05). After treatment, CCr in the treatment group was (56.3 ± 5.01) mL/min, which was significantly increased compared with that before treatment ( t = -8.56, P < 0.05). After treatment, Cys-C, CRP, and UAER in the control group were (2.17 ± 0.04) mg/L, (4.66 ± 1.47) mg/L, and (103.93 ± 22.62) μg/min, respectively, which were significantly decreased compared with those before treatment ( t = 4.05, 5.00, 2.24, all P < 0.05). After treatment, CCr in the control group was (45.9 ± 4.9) mL/min, which was significantly increased compared with that before treatment ( t = -3.98, P < 0.05). There were significant differences in Cys-C, UAER, and CCr between the treatment and control groups ( t = -7.42, -2.29, 7.82, all P < 0.05). Conclusion:Tanzhuo Decoction for the treatment of early diabetic nephropathy in patients with type 2 diabetes mellitus has a definite effect. It can effectively reduce levels of Cys-C and UAER, reduce inflammatory reactions, improve kidney function, and delay the progression of kidney injury.
9.Analysis of Risk Factors for Impaired Branch Perfusion After Percutaneous Coronary Intervention of Coronary True Bifurcation Lesions Based on Quantitative Flow Ratio
Yubo LONG ; Ting ZHOU ; Hongwei PAN ; Yuanyuan LI ; Changlu WANG ; Yu ZHANG ; Hu HU ; Zun HU ; Jingjing RONG
Chinese Circulation Journal 2024;39(6):554-561
Objectives:Present study aimed to use quantitative flow ratio based on Murray's law to analyze the risk factors of impaired side branches perfusion without naked eye visible slowing of blood flow in branches after interventional treatment of true bifurcation lesions of the coronary arteries. Methods:A total of 211 patients with non-left main coronary artery true bifurcation coronary artery disease who underwent percutaneous coronary intervention(PCI)in Hunan Provincial People's Hospital from June 2022 to September 2023 were continuously enrolled,with a total of 234 bifurcation lesions.The general clinical indicators,anatomical characteristics of coronary artery bifurcation lesions,branch protection methods,postoperative branch TIMI blood flow and other data were collected,and quantitative flow ratio(μQFR)was measured for postoperative branch blood vessels.Post-PCI μQFR<0.8 was considered as impaired branch perfusion and was included in the postoperative impaired branch perfusion group(n=51,53 branch lesions).Patients with μQFR≥0.8 were included in the postoperative normal branch perfusion group(n=160,181 branch lesions).Multivariate Logistic regression analysis was used to evaluate the effects of various clinical and anatomical factors on branch perfusion after PCI. Results:The post-PCI branch flow grading of all patients was TIMI grade Ⅲ.The postoperative branch μQFR of 53 vessels(22.6%)in the group with impaired postoperative branch perfusion was 0.70±0.10,and 0.93±0.05 in the group with normal postoperative branch perfusion,and the difference between the two groups was statistically significant(P<0.001).Compared with the postoperative group with normal branch perfusion,the postoperative group with impaired branch perfusion was featured with an elevated branch lesion length,branch reference diameter,postoperative branch opening diameter stenosis rate,postoperative branch narrowest lumen diameter stenosis rate,and a lower main branch-to-branch diameter ratio,preoperative branch narrowest lumen diameter stenosis rate,and preoperative main branch μQFR,all of which were statistically significant(all P<0.05).The postoperative branch opening diameter stenosis rate(r=-0.490,P<0.001),postoperative branch narrowest lumen diameter stenosis rate(r=-0.788,P<0.001),preoperative branch narrowest lumen diameter stenosis rate(r=-0.280,P<0.001),branch narrowest lumen diameter(r=-0.469,P<0.001),branch lesion length(r=-0.157,P=0.016)were negatively correlated with postoperative branch μQFR,and branch reference diameter(r=0.173,P=0.008),main branch/side branch diameter ratio(r=0.194,P=0.003),and branch opening diameter(r=0.328,P<0.001)were positively correlated with postoperative branch μQFR,and none of them were significantly correlated with clinical baseline data(all P>0.05).Multifactorial logistic regression analysis showed that following four factors were independent risk factors for impaired branch perfusion:postoperative stenosis of the narrowest branch lumen diameter(OR=1.228,95%CI:1.144-1.318,P<0.001),postoperative stenosis of the branch opening diameter(OR=1.110,95%CI:1.055-1.168,P<0.001),postoperative stenosis of the narrowest lumen diameter of the main branch(OR=1.115,95%CI:1.042-1.192,P=0.001),and length of the branch lesion(OR=1.121,95%CI:1.021-1.231,P=0016). Conclusions:Some of the patients whose branch flow reached TIMI grade Ⅲ after PCI are still faced the risk of hemodynamical impairment and should be functionally evaluated after PCI.The postoperative stenosis rate of the narrowest branch lumen diameter,postoperative stenosis rate of the branch opening diameter,postoperative stenosis rate of the narrowest lumen diameter of the main branch,and branch lesion length are the risk factors of branch perfusion impairment after PCI for coronary bifurcation lesions.
10.A Preliminary Study on the Efficacy of Percutaneous Coronary Intervention for Complex Left Main Stem Combined With Chronic Total Occlusion of the Right Coronary Artery
Zun HU ; Hu HU ; Yubo LONG ; Junshan LI ; Jingjing RONG ; Jin HE ; Changlu WANG ; Yu ZHANG ; Jianqiang PENG ; Hongwei PAN
Chinese Circulation Journal 2024;39(6):562-567
Objectives:to analyze the efficacy of percutaneous coronary intervention(PCI)for complex left main(LM)lesions combined with chronic total occlusion(CTO)of the right coronary artery. Methods:Ninety patients with complex left main lesions hospitalized in Hunan Provincial People's Hospital from January 2019 to December 2022 were consecutively included.According to the coronary angiographic vascular lesions,patients were divided into complex left main lesions combined with right coronary artery CTO(observation group,n=30)and complex left main lesions without right coronary artery CTO(control group,n=60).The baseline clinical data,intraoperative conditions,angiographic results,and postoperative follow-up results of the patients were analyzed and compared between the two groups. Results:Fifty-eight(64.4%)out of the 90 patients were male.There was no statistically significant difference between the two groups in terms of baseline clinical data(all P>0.05),left main lesion condition(P=1.000),left main calcification condition(P=0.249),and preoperative TIMI flow grading(P=1.000).In the comparison between observation group and the control group,intraoperative occurrence of no-reflow(3.3%vs.5.0%,P=1.000),hypotension(10.0%vs.8.3%,P=1.000),pericardial effusion(3.3%vs.0%,P=0.333),the percentage of intravascular ultrasound(IVUS)use(86.7%vs.90.0%,P=0.635),and the use of circulatory assist device(P=0.699),and the proportion of intraoperative coronary spinning(26.7%vs.21.7%,P=0.597)were all similar between the two groups.The median follow-up time was 14.50(11.83,15.85)months,and the differences in the incidence of major adverse cardiovascular events(MACE)such as recurrent angina,acute myocardial infarction,rebleeding,readmission for heart failure,and cardiac death(31.0%vs.32.1%,P=1.000)were not statistically significant between the observation group and the control group. Conclusions:PCI revascularization may be a viable approach for elderly patients with complex LM lesions with multiple underlying disease,and combined right coronary artery CTO,intolerance and reluctance to CABG.

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