1.Key Information Research and Modern Clinical Application of Xiaofengsan
Weilu NIU ; Mengjie YANG ; Chengqi LYU ; Cuicui SHEN ; Ningli WANG ; Huangchao JIA ; Liyun WANG ; Xuewei LIU ; Mingsan MIAO ; Xiaomeng WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):238-249
Employing bibliometric methods and adhering to principles of textual research, this study systematically investigated prescription source, formula name, composition evolution, dose evolution, origin, processing, ancient and modern applications of Xiaofengsan. Xiaofengsan, also known as Renshen Xiaofengsan and Chantui Xiaofengsan, was first recorded in the Taiping Huimin Hejijufang(hereafter referred to as Jufang) of the Southern Song dynasty. The formula composition included Schizonepetae Spica, Glycyrrhizae Radix et Rhizoma, Chuanxiong Rhizoma, Notoptery Rhizoma et Radix, Bombyx Batryticatus, Saposhnikoviae Radix, Poria, Cicadae Periostracum, Pogostemonis Herba, Ginseng Radix et Rhizoma, Magnoliae Officinalis Cortex and Citri Reticulatae Pericarpium, a total of 12 medicinal materials. In terms of the evolution of formula composition, formulas across dynasties largely aligned with those recorded in Jufang, with only minor variations in application. The results of the formula dosage research indicated that one dose of medication in Jufang corresponded to the following modern dosages:Schizonepetae Spica of 82.6 g, Glycyrrhizae Radix et Rhizoma of 82.6 g, Chuanxiong Rhizoma of 82.6 g, Notoptery Rhizoma et Radix of 82.6 g, Bombyx Batryticatus of 82.6 g, Saposhnikoviae Radix of 82.6 g, Poria of 82.6 g, Cicadae Periostracum of 82.6 g, Pogostemonis Herba of 82.6 g, Ginseng Radix et Rhizoma of 82.6 g, Magnoliae Officinalis Cortex of 20.65 g and Citri Reticulatae Pericarpium of 20.65 g, the origins of all the constituent drugs were consistent with the 2020 edition of Pharmacopoeia of the People's Republic of China. The results of the investigation into the decoction method indicated that the aforementioned drugs should be finely ground into powder(pass through the No.5 sieve), and 8.26 g was taken for each dose, which was taken with the clear liquid obtained by steeping tea leaves in boiling water for several minutes. This mixture was administered three times daily, 30 min after meals. The ancient functional indications of this formula mainly involved dispelling wind-heat, eliminating pathogenic factors and regulating the middle Jiao. It primarily treated all wind-heat syndromes manifesting as skin diseases, predominantly affecting the upper body, especially the head and face. The diseases involved in modern applications were mostly dermatological diseases, including urticaria, eczema, atopic dermatitis and others. In this paper, by combing the relevant ancient literature, the key information of Xiaofengsan was textual researched, in order to provide reference for the modern application and development of this formula.
2.Screening of the Monkeypox Virus Key Proteins Regulating Inflammation Based on NOD-like Receptor Family Pyrin Domain Containing 3 Inflammasome Signaling
Mingxing LUO ; Chenchen PEI ; Liyun GONG ; Xinlei ZHENG ; Jun QIAN ; Quan LIU ; Jun ZHENG
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(2):259-268
ObjectiveTo screen and identify the key proteins of Monkeypox Virus (MPXV) that regulate the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. MethodsUsing the genomic sequence of the domestic epidemic strain of MPXV as a template to construct an expression plasmid library containing 82 viral genes. To screen for viral genes that significantly inhibit the production of interleukin-1β (IL-1β), the expression plasmids encoding NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteine protease-1(Caspase-1), and pro-IL-1β-Luc reporter gene were co-transfected into HEK293T cells, along with the viral gene plasmids. The signal of the NLRP3 inflammasome was monitored by measuring luciferase activity in the cell supernatant. The inhibition effects were further confirmed by Western Blot (WB), and the interactions between the viral proteins and the components of the NLRP3 inflammasome were determined by Co-immunoprecipitation (Co-IP). Additionally, site-directed mutagenesis was performed to identify the specific interaction motifs. ResultsAn NLRP3 inflammasome signal screening system were successfully established and optimized. Screening the MPXV plasmids using this system revealed that B10R most significantly inhibits IL-1β maturation in a dose-dependent manner (P<0.000 1). Further characterization demonstrated that B10R interacts directly with pro-IL-1β. Notably, the C-terminal motif mutant of B10R (B10Rmut-C, T96A, Y97A, I98A) lost its ability to interact with pro-IL-1β. ConclusionsThe MPXV B10R protein interacts with pro-IL-1β, thereby inhibiting the IL-1β maturation. The C-terminal motif (T96, Y97, I98) of the B10R protein is crucial for function. This study provides important clues and a foundation for further elucidating the molecular mechanisms by which MPXV regulates the NLRP3 inflammasome.
3.Antiviral research and drug development targeting the solute carrier superfamily (SLCs)
Liyun DING ; Dongyu LIU ; Ting ZHOU ; Wei PAN ; Jinlei BIAN
Journal of China Pharmaceutical University 2026;57(4):411-421
The solute carrier superfamily (SLCs), comprising over 450 transporter proteins, plays pivotal roles in nutrient absorption, transmembrane transport regulation, and immunometabolism, and is intimately linked to various diseases. This review systematically summarizes the latest progress of SLCs in the antiviral field, focusing on three classification mechanisms: firstly, SLCs function as critical receptors or cofactors for viral entry, mediating viral attachment to and fusion with host cell membranes; secondly, SLCs modulate host innate immune signaling pathways, thereby positively or negatively regulating antiviral immune responses; and thirdly, SLCs serve as transmembrane transporters for antiviral agents, determining their absorption, distribution, and metabolism, which significantly influence therapeutic efficacy. Furthermore, host-targeted antiviral strategies based on these mechanisms and the chemical structures of relevant bioactive molecules are systematically elucidated. Finally, the potential of targeting SLCs for novel antiviral drug development and the associated druggability challenges are discussed in depth, providing a theoretical basis for the molecular design and development of next-generation antiviral agents.
4.Exploration of the Body-Mind Harmonization Strategy in Treatment of Psoriasis with Anxiety and Depression Based on the "Depression-Blood-Spirit" Transformation Theory
Wenjie WANG ; Huaiwei LIU ; Shuying ZHA ; Liyun SUN
Journal of Traditional Chinese Medicine 2025;66(13):1395-1399
It is believed that psoriasis with anxiety and depression presents the pathogenesis of "depression-blood-spirit", with qi depression as the initiating factor, and that the disease mechanism is understood from three aspects, including "qi depression transforming into fire, blood stasis disturbing the spirit", "depressed fire consuming yin, causing blood dryness and spirit agitation", "qi stagnation leading to blood stasis, resulting in obstructed channels and spirit depression". The treatment proposes three methods, firstly, cooling blood and resolving depression, secondly, nourishing blood and soothing depression, and thirdly, activating blood and regulating depression, which applies prescriptions as modified Liangxue Huoxue Decoction (凉血活血汤) combined with Zhiqiao (Poncirus trifoliata), Foshou (Citrus medica), and Lianqiaoxin (Forsythia suspensa) for cooling blood and resolving depression; modified Yangxue Jiedu Decoction (养血解毒汤) combined with Baishao (Paeonia lactiflora Pall), Zhenzhumu (Margaritifera Concha), and calcined Muli (Ostreidae) for nourishing blood and soothing depression; and modified Huoxue Sanyu Decoction (活血散瘀汤) combined with Yujin (Curcuma aromatica) and Hehuanpi (Albiziae Cortex) for activating blood and regulating depression. These three methods integrate the "depression-blood-spirit" pathogenesis to achieve harmony of body and spirit by clearing blood heat, nourishing yin and blood, and removing blood stasis, complemented with medicinals that soothing the liver, calming the spirit, and regulating depression.
5.Key Information Research and Modern Clinical Application of Famous Classical Formula Yanghetang
Weilu NIU ; Mengjie YANG ; Chengqi LYU ; Shunxi WANG ; Ziwen WANG ; Huangchao JIA ; Liyun WANG ; Xuewei LIU ; Mingsan MIAO ; Xiaomeng WANG ; Yawei YAN ; Chunyong LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):238-246
Through data collection and collation combined with bibliometrics, this study conducted a series of textual research on Yanghetang, such as the name and origin, the evolution of prescription composition and modern clinical application. Yanghetang was first recorded in Bencao Yidu of WANG Ang in the Qing dynasty. In addition to Yanghetang, there were 3 bynames of Jiawei Yanghetang, Quanshengji Yanghetang and Zhenjun Yanghetang. Regarding the composition of the formula, a total of 4 versions of Yanghetang were collected. The first version is the 5 medicines version of Cervi Cornus Colla, Rehmanniae Radix Praeparata, Cinnamomi Cortex, Zingiberis Rhizoma and Ephedrae Herba in Bencao Yidu. The second version is the 7 medicines version of Waike Zhengzhi Quanshengji, changing Zingiberis Rhizoma to Zingiberis Rhizoma Praeparatum Carbonisata(ZRPC) and adding Sinapis Semen and Glycyrrhizae Radix et Rhizoma(GRR) on the basis of Bencao Yidu, and most of the Yanghetang is of this version. The third version is the 6 medicines version of Wushi Yifang Huibian, that is, on the basis of Bencao Yidu, Zingiberis Rhizoma is changed into ZRPC, and Sinapis Semen is added. The fourth version is the 6 medicines version in Yifang Jiedu, that is, on the basis of Bencao Yidu, Zingiberis Rhizoma is changed into Zingiberis Rhizoma Praeparatum, and GRR Praeparata cum Melle is added. Regarding the dose of Yanghetang, the doses of the medicines in Waike Zhengzhi Quanshengji was converted into the modern doses as follows:37.3 g of Rehmanniae Radix Praeparata, 1.87 g of Ephedrae Herba, 11.19 g of Cervi Cornus Colla, 7.46 g of Sinapis Semen, 3.73 g of Cinnamomi Cortex, 3.73 g of GRR, and 1.87 g of ZRPC. The origins of the above medicines are consistent with the 2020 edition of Chinese Pharmacopoeia. The processing specification of Rehmanniae Radix Praeparata is steaming method, ZRPC is ginger charcoal, Sinapis Semen is the fried products, and the rest of the medicines are raw products. The decoction method was verified by the decoction method in Chonglou Yuyao, which is similar in the time, and it is recommended that the above medicines should be added with 600 mL of water, decocted to 100 mL, and taken warmly 30 min after meal. For each dose, it is recommended to use 1-3 doses per day according to the doctor's advice in combination with clinical practice. The diseases involved in the ancient applications involved 42 diseases in 11 departments, including orthopedics, dermatology and gynecology, which were dominated by Yin-cold syndrome. However, the diseases involved in modern research also include 148 related diseases in 10 departments, such as orthopedics, obstetrics and gynecology, which is consistent with the ancient books. In recent years, the research hotspots of Yanghetang have focused on more than 10 fields, including osteoblasts, malignant tumors, wound healing, traditional Chinese medicine fumigation and so on, which are widely used. It is suitable for comprehensive research and development because of its rational formula composition, clear origin, processing and decoction method, and wide clinical application.
6.Exploration on the Influence of Jiedu Huayu Granules on the Clinical Efficacy of Patients with Liver Failure from the Perspective of Endoplasmic Reticulum Stress and Inflammatory Response
Wu XIE ; Minming YI ; Liyun LIN ; Shousong YANG ; Huihua LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(3):593-599
Objective To investigate the clinical efficacy of Jiedu Huayu Granules(mainly composed of the Chinese herbal medicines of Artemisiae Scopariae Herba,Rhei Radix et Rhizoma,Hedyotis Diffusae Herba,Paeoniae Radix Rubra,Curcumae Radix and Acori Tatarinowii Rhizoma)in the treatment of liver failure(LF)and to observe their effects on endoplasmic reticulum stress and inflammatory response.Methods From May 2022 to May 2024,a total of 102 patients with LF of toxin-heat-stasis stagnation syndrome were collected from Hepatology Department of Yulin Traditional Chinese Medicine Hospital,and were divided into the control group and the study group by using a table of random numbers,with 51 cases in each group.The two groups were given comprehensive western medicine treatment for inhibiting inflammatory response and protecting liver,regulating intestinal microecology,counteracting viral infection,decreasing transaminase level and treating jaundice.Additionally,the study group was treated with Jiedu Huayu Granules.The course of treatment for the two groups covered eight weeks.Before and after treatment,the two groups were observed in the changes of traditional Chinese medicine(TCM)syndrome scores,biochemical indicators,inflammatory factors and endoplasmic reticulum stress indicators.After treatment,the clinical efficacy and the incidence of adverse reactions of the two groups of patients were compared.Results(1)After eight weeks of treatment,the total effective rate of the study group was 86.27%(44/51)and that of the control group was 68.63%(35/51),and the intergroup comparison(tested by chi-square test)showed that the efficacy of the study group was significantly superior to that of the control group(P<0.05).(2)After treatment,the TCM syndrome scores in both groups were significantly decreased compared with those before treatment(P<0.01),and the decrease in the study group was significantly superior to that in the control group(P<0.05).(3)After treatment,the serum levels of biochemical indicators of total bilirubin(TBIL),total bile acid(TBA),alanine transaminase(ALT),and aspartate transaminase(AST)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group was significantly superior to that in the control group(P<0.05 or P<0.01).(4)After treatment,the serum levels of inflammatory factors of interleukin 6(IL-6),tumor necrosis factor α(TNF-α),transforming growth factor β(TGF-β),and C-reactive protein(CRP)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group was significantly superior to that in the control group(P<0.01).(5)After treatment,the levels of endoplasmic reticulum response indicators such as glucose-regulated protein 78(GRP78),CCAAT enhancer-binding protein homologous protein(CHOP),and cysteine aspartic acid specific protease 12(caspase-12)in the two groups were significantly decreased compared with those before treatment(P<0.05),and the decrease in the study group were significantly superior to those in the control group(P<0.01).(6)The incidence of adverse reactions in the study group was 15.69%(8/51)and that in the control group was 17.65%(9/51),and the comparison between the two groups showed that the difference was not statistically significant(P>0.05).Conclusion The combined use of Jiedu Huayu Granules with comprehensive western medicine treatment exerts certain efficacy in treating patients with LF of toxin-heat-stasis stagnation syndrome.The combined therapy is effective on relieving clinical symptoms and improving liver function,and is helpful for relieving endoplasmic reticulum stress and inflammatory response.
7.Relationships of serum procalcitonin,α1-acid glycoprotein and cluster of differentiation 64 index with prognosis in patients with Candida infection after lung cancer surgery
Huifeng ZHANG ; Tianwen HOU ; Yunzhou ZHENG ; Liyun AN ; Jing CHEN ; Ye LIU ; Juntao LI ; Rongying TIAN
Journal of Clinical Medicine in Practice 2025;29(10):52-56
Objective To investigate the relationships of serum procalcitonin(PCT),α1-acid glycoprotein(α1-AGP)and cluster of differentiation 64(CD64)index with the severity of illness and prognosis in patients with pulmonary Candida infection after lung cancer surgery.Methods A pro-spective study was conducted in 120 patients with pulmonary Candida infection after lung cancer sur-gery as infection group,and 60 patients without infection after lung cancer surgery in the same period were selected as control group.Baseline characteristics and laboratory indicators were compared be-tween the two groups.Differences in serum PCT,α1-AGP,CD64 index in peripheral blood,and plasma 1,3-β-D glucan test(BG)were compared among patients with different severities of illness and prognoses in the infection group.The relationship between serum indicators and prognosis as well as their predictive efficiencies were analyzed,and external validation was performed.Results The levels of serum PCT,α1-AGP,CD64 index in peripheral blood,and plasma BG in the infection group were significantly higher than those in the control group(P<0.05).In the infection group,the levels of serum PCT,α1-AGP,and CD64 gradually decreased significantly in patients with severe,moderate,and mild illness,and the levels of these indicators in dead patients were significantly higher than those in surviving patients(P<0.05).Survival curve analysis showed that patients with low levels of serum PCT,α1-AGP,and CD64 index before treatment had higher survival rates than those with high levels of these indicators.Receiver operating characteristic(ROC)curve analysis revealed that the combined prediction of PCT,α1-AGP,and CD64 index for mortality was superior to individual indicators or their pairwise combinations,and external validation confirmed the good predictive efficacy of the combined prediction.Conclusion PCT,α1-AGP,and CD64 index are closely related to pulmonary Candida infection after lung cancer surgery,and the combined detection of these indicators has certain predictive value for 28-day prognosis.
8.Multi-center clinical study on the efficacy and safety of combined lienal polypeptide injection therapy in children with Mycoplasma pneumoniae pneumonia
Qi CHENG ; Yunxiao SHANG ; Han ZHANG ; Jiujun LI ; Ning CHEN ; Lishen SHAN ; Nan YANG ; Lihua NING ; Xuemei BAI ; Jianhua LIU ; Yuling HAN ; Jichun WANG ; Jing LI ; Yong FENG ; Liyun LIU ; Li CHEN ; Si LIU ; Qinzhen ZHANG ; Jia ZHENG ; Fengchao LI ; Sukun LU ; Yun ZHANG ; Xiaoyi CHAI
International Journal of Pediatrics 2025;52(3):204-210
Objective:To observe the efficacy and safety of combined lienal polypeptide injection therapy in the treatment of Mycoplasma pneumoniae pneumonia(MPP)in children aged 3 to 14 years old in multiple clinical centers.Methods:A randomized,controlled,multi-center clinical study design was adopted.A total of 240 hospitalized children aged 3 to 14 years old with MPP from 7 hospitals from September 1,2023 to January 31,2024 were included.According to the severity of pneumonia,they were divided into the mild MPP group with 80 cases and the severe MPP/refractory MPP(SMPP/RMPP)group with 160 cases,and then randomly divided into the control group and the experimental group at a ratio of 1 ∶1,using the random number table method.After screening,subjects entered a treatment period of 5 to 7 days.The control group was treated with azithromycin,while the experimental group was treated with azithromycin plus lienal polypeptide injection .The recovery of lung CT,length of hospital stay,duration of fever,cough score,whether mild cases developed into severe or refractory cases,duration of hormone use,use of intravenous immunoglobulin(IVIG),bronchoscopy treatment,and immune function were observed between the two groups to evaluate the efficacy of lienal polypeptide injection.Adverse events after medication,vital signs,blood routine,urine routine,liver function,myocardial enzymes,renal function,and electrocardiogram were observed to evaluate the safety. Results:A total of 231 subjects have completed the trial in the 7 hospitals,including 118 cases in the experimental group and 113 cases in the control group.Main observation index:the rate of lung CT aggravation in the experimental group was lower than that in the control group(2.6% vs 15.3%, P<0.01),and the difference was statistically significant.Secondary indexes:there were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and immunoglobulin between the two groups(all P>0.05).However,the rate of cases of plastic bronchitis(PB)found under bronchoscopy in the experimental group was lower than that in the control group(0 vs 18.8%, P=0.03),and the difference was statistically significant.Among the mild MPP(72 cases),there were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and the improvement rate of lung CT between the two groups(all P>0.05).However,compared with the control group,the rate of cases developing into SMPP/RMPP in the experimental group was less(24.3% vs 48.6%, P=0.03),and the difference in IgG before and after treatment was small[0.53(-0.04,1.18)g/L vs 1.33(0.48,2.25)g/L, P=0.01].Among the SMPP/RMPP cases(159 cases),the rate of cases of PB found under bronchoscopy in the experimental group was less than that in the control group(0 vs 20%, P=0.04),and the rate of cases with aggravated lung CT in the experimental group was less than that in the control group(1.3% vs 19.5%, P<0.01),and the improvement rate of lung CT in the experimental group was higher than that in the control group(88.8% vs 75.3%, P=0.03),with statistically significant differences.There were no statistically significant differences in the length of hospital stay,duration of fever,cough score,duration of hormone use,whether IVIG treatment was used,the number of bronchoscopy treatment cases,and immunoglobulin between the two groups(all P>0.05).Two cases in the experimental group developed rashes,which improved after the drug was discontinued.There were no serious adverse reactions such as abnormal vital signs like dyspnea and cyanosis due to the use of lienal polypeptide injection.There were no obvious changes in blood routine,liver function,myocardial enzymes,renal function,electrocardiogram,and urine routine values before and after medication compared with the baseline. Conclusion:The combined use of lienal polypeptide injection in the treatment of MPP in children can reduce the probability of the transformation from mild cases to SMPP/RMPP,reduce the rate of aggravation of the image findings,promote the absorption of lung inflammation,reduce the rate of PB found under bronchoscopy,and has good safety.
9.Research progress on the relationship between the photobiomodulation and amblyopia
Shuxian HU ; Mei LIU ; Jingjing DONG ; Yang YANG ; Li LIU ; Xuan MA ; Liyun GUO
International Eye Science 2025;25(9):1431-1435
Amblyopia is a common visual development disorder and is the main cause of monocular vision impairment in children and adults. Photobiomodulation(PBM), a non-invasive treatment method, has gradually gained attention in the field of ophthalmology. This paper begins with the macroscopic manifestation of light on the animal model of amblyopia. Additionally, it discusses the pathological changes of the amblyopic retina and the human eye's central nervous system, as well as the influence and mechanism of PBM on the visual perception and processing system and its chemical effect on the visual system through dopamine and melatonin. It examines its mechanism of action, current clinical application status, and future development direction in order to provide new ideas and theoretical foundation for amblyopia treatment.
10.Historical Evolution and Modern Clinical Application of Huoxiang Zhengqisan
Weilu NIU ; Mengjie YANG ; Chengqi LYU ; Cuicui SHEN ; Congcong LI ; Huangchao JIA ; Liyun WANG ; Xuewei LIU ; Mingsan MIAO ; Xiaomeng WANG ; Yawei YAN ; Chunyong LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):156-167
In this study, bibliometric methods were used to systematically investigate the name and origin, the evolution of prescription composition, dose evolution, origin and processing method, decoction method, ancient application, modified application, modern application and other information of Huoxiang Zhengqisan. After research, Huoxiang Zhengqisan, also known as Huoxiang Zhengqitang, was first recorded in Taiping Huimin Hejijufang. The original formula is composed of 41.3 g of Arecae Pericarpium, 41.3 g of Angelicae Dahuricae Radix, 41.3 g of Perilla frutescens(actually Perillae Folium), 41.3 g of Poria, 82.6 g of Pinelliae Rhizoma, 82.6 g of Atractylodis Macrocephalae Rhizoma, 82.6 g of Citri Reticulatae Pericarpium(actually Citri Exocarpium Rubbum), 82.6 g of Magnoliae Officinalis Cortex, 82.6 g of Platycodonis Radix, 123.9 g of Pogostemonis Herba, and 103.25 g of Glycyrrhizae Radix et Rhizoma. In this formula, Magnoliae Officinalis Cortex is processed according to the specifications for ginger-processed products, Glycyrrhizae Radix et Rhizoma is processed according to the specifications for stir-fried products, and other herbs are used in their raw products. The botanical sources of the herbs are consistent with the 2020 edition of Pharmacopoeia of the People's Republic of China. The above herbs are ground into a fine powder with a particle size passing through a No. 5 sieve. For each dose, take 8.26 g of the powdered formula, add 300 mL of water, along with 3 g of Zingiberis Rhizoma Recens and 3 g of Jujubae Fructus, and decoct until reduced to 140 mL. The decoction should be administered hot, with three times daily. To induce sweating, the patient should be kept warm under a quilt, and an additional dose should be prepared and taken if needed. This formula is traditionally used to relieve the exterior and resolve dampness, regulate Qi and harmonize the middle, which is mainly used to treat a series of diseases of digestive and respiratory systems. However, potential adverse reactions, including allergies, purpura and disulfiram-like reactions, should be considered during clinical use. Huoxiang Zhengqisan features a rational composition, extensive clinical application, and strong potential for further research and development.

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