1.Effect of Liangxue Tuizi Formula (凉血退紫方) on RAF/MEK/ERK Pathway in Skin Tissue and Serum NETs Biomarkers in Henoch-Schönlein Purpura Model Rats with Blood Heat Syndrome
Yingying JIANG ; Manxiang YANG ; Zhenhua YUAN ; Leying XI ; Mingyang CAI ; Diya MA ; Yifan LI ; Yuhang NIU ; Runze LIU ; Jiawen CAO ; Xilin CHEN ; Xianqing REN
Journal of Traditional Chinese Medicine 2025;66(23):2475-2483
ObjectiveTo investigate the potential mechanism of Liangxue Tuizi Formula (凉血退紫方, LXTZF) in treating Henoch-Schönlein Purpura (HSP) by examining its regulatory effect on neutrophil extracellular trap (NETs) dysregulation via the rapidly accelerated fibrosarcoma kinase (RAF)/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodsSeventy Wistar rats were randomly allocated into a blank control group (n=14) and a modeling group (n=56). Rats in the modelling group underwent an eight-week modelling period to establish HSP rat models with blood-heat syndrome via modified ovalbumin (OVA) induction method combined with oral administration of heat-property Chinese herbal medicine. Fifty successfully modeled rats were subsequently randomly divided into five groups (n=10 per group), model group, compound glycyrrhizin group, LXTZF group, RAF inhibitor group, and LXTZF + RAF agonist group. Additionally, 10 rats were selected from the original blank control group for the final experiment. From the 11th week of modelling, rats in the blank control group and the model group received 1 ml/(100 g·d) ultrapure water via oral administration, in addition to 0.5 ml/(kg·d) 0.9% sodium chloride solution via intraperitoneal injection. The LXTZF group and the compound glycyrrhizin group received 7.5 g/(kg·d) LXTZF granule suspension via gavage, 13.5 mg/(kg·d) compound glycyrrhizin suspension via gavage, respectively. The RAF inhibitor group received 1 mg/(kg·d) GW5074 suspension via intraperitoneal injection and ultrapure water via oral administration; the LXTZF + RAF agonist group received 7.5 g/(kg·d) LXTZF granule suspension via gavage and 1 mg/(kg·d) paclitaxel suspension via intraperitoneal injection. All administrations were performed once daily for 4 weeks. After intervention, skin tissue histopathology was examined by hematoxylin and eosin (H&E) staining, immunoglobulin A (IgA) deposition was assessed via immunofluorescence, serum levels of neutrophil elastase (NE), tumor necrosis factor-α (TNF-α), and vascular cell adhesion molecule-1 (VCAM-1) were measured using enzyme-linked immunosorbent assay (ELISA), serum myeloperoxidase (MPO) level was determined by a colorimetric assay; the mRNA expression levels of RAF, MEK, and ERK in skin tissue were detected by real-time quantitative polymerase chain reaction (RT-qPCR); and the protein expression of RAF, MEK, ERK, as well as phosphorylated MEK (p-MEK) and phosphorylated ERK (p-ERK), were analyzed by Western Blot. ResultsSkin tissue in the blank control group rats remained normal, whereas the model group exhibited neutrophil infiltration and haemorrhage with red blood cell rupture. In all drug intervention groups, neutrophil infiltration and haemorrhagic exudation reduced markedly, with LXTZF group demonstrating the most pronounced improvement. Compared with the blank control group, rats in the model group exhibited enhanced IgA fluorescence intensity in skin tissue, elevated serum levels of NE, MPO, TNF-α and VCAM-1, increased mRNA expression of RAF, MEK, ERK1 and ERK2, as well as heightened RAF protein levels and p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with the model group, the drug intervention groups exhibited reduced IgA fluorescence intensity in skin tissue, along with decreased serum levels of NE, MPO, TNF-α, and VCAM-1 (P<0.05). In LXTZF group and RAF inhibition groups, reduced mRNA expression of RAF, MEK, ERK1, and ERK2 was observed in rat skin tissue, alongside decreased RAF protein levels and reduced p-MEK/MEK and p-ERK/ERK ratios (P<0.05). Compared with LXTZF + RAF agonist group, the compound glycyrrhizin group, LXTZF group, and RAF inhibitior group exhibited reduced IgA fluorescence intensity in skin tissue, decreased serum NE, MPO, TNF-α, and VCAM-1 levels, and decreased MEK mRNA expression and p-MEK/MEK ratio (P<0.05). ConclusionThe potential mechanism by which LXTZF treats Henoch-Schönlein purpura with blood heat syndrome may involve blocking the RAF/MEK/ERK signaling pathway in skin tissue, and suppressing excessive formation of NETs, thereby reducing IgA deposition in dermal microvessels and attenuating systemic inflammatory responses.
2.Expert consensus on antiviral therapy of COVID-19
Fujie ZHANG ; Zhuo WANG ; Quanhong WANG ; Qing MAO ; Jinsong BAI ; Hanhui YE ; Jia TIAN ; Tianxin XIANG ; Jihong AN ; Zujiang YU ; Wenjie YANG ; Xingxiang YANG ; Xiaoju ZHANG ; Jie ZHANG ; Lina ZHANG ; Xingwang LI ; Jiabin LI ; Manxiang LI ; Zhiwei LI ; Hourong ZHOU ; Yi SHI ; Xiaoling XU ; Xiaoping TANG ; Hong TANG ; Xixin YAN ; Wenxiang HUANG ; Chaolin HUANG ; Liang DONG ; Baosong XIE ; Jiandong JIANG ; Bin XIONG ; Xuemei WEI ; Jifang SHENG ; Ronghua JIN
Chinese Journal of Clinical Infectious Diseases 2023;16(1):10-20
COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.
3.Effect of Catalpol on the Activity and Apoptosis of Osteoclast in the Osteoblasts-osteoclasts Co-culture Sys-tem and Its Mechanism Study
Manxiang LAI ; Xia CHEN ; Juan FENG ; Lixia HE ; Li YANG
China Pharmacy 2016;27(10):1318-1321
OBJECTIVE:To study the effect of catalpol on the activity and apoptosis of osteoclasts (OC) in the osteoblasts (OB)-OC co-culture system and its mechanism. METHODS:OB and OC were isolated respectively from the SD rats of 1-3 days and 5-7 days old to establish OB-OC co-culture system. After treated with 0(blank control),0.05,0.5,5,50 and 100 mg/L catal-pol for 48,72 and 96 h,the number of bone absorption lacuna for OC was observed by inverted microscope to reflect OC activity. After treated with 0(blank control)and 0.05 mg/L catalpol for 48,72 and 96 h,the activity of tartrate resistant acid phosphatase (TRACP)in OC was detected,and the apoptosis rate of OC was calculated. After treated with 0(blank control)and 0.05 mg/L ca-talpol,mRNA expression of osteoprotegerin(OPG)in OB was detected. RESULTS:In OB-OC co-culture system,the number of bone absorption lacuna in 0.05-50 mg/L catalpol groups was significantly lower than blank control group(P<0.01),indicating ca-talpol could inhibit OC activity,especially 0.05 mg/L catapol. Compared with blank control,0.05 mg/L catapol lowered the activity of TRACP but increased the apoptosis rate of OC(P<0.05);mRNA expression of OPG was up-regulated in OB(P<0.01). CON-CLUSIONS:In OB-OC co-culture system,catalpol can inhibit the activity of OC and induce the apoptosis of OC,and its mecha-nism may be associated with the mRNA expression up-regulation of OPG in OB.
4.Effect of catalpol on activity of osteoblasts/osteoclasts and osteoblast ERα/βmRNA expression in osteoblast-osteoclast co-culture system
Manxiang LAI ; Li YANG ; Ronghua ZHANG
Chinese Journal of Pathophysiology 2015;(7):1242-1246
[ ABSTRACT ] AIM: To investigate the effect of catalpol on the activity of osteoblasts ( OB ) and osteoclasts ( OC) , and OB estrogen receptor ( ER) α/βmRNA expression in the OB-OC co-culture system.METHODS: OB and OC were isolated from the SD rats of 1 and 5 days old.In the OB-OC co-culture system, different concentrations of catalpol including low dosage (0.05 , 0.1, 0.5 and 1 mg/L), middle dosage (2, 5 and 10 mg/L), and high dosage (20, 50 and 100 mg/L) were added into the culture medium to detect the changes of OB proliferation by MTT assay.The catalpol at maximal dosage was added to OB section to detect the alkaline phosphatase ( ALP) activity of OB by pNPP method.The mRNA expression of ERα/βin the OB treated with catalpol in the co-culture system was detected by RT-PCR.The catalpol at maximal dosage was added to OC group to detect the activity of OC by microscopy and tartrate-resistantacid phosphatase ( TRAP) activity detection.RESULTS:In 0.05~2 mg/L catalpol groups, the proliferation of OB was significantly in-creased as compared with control group in the co-culture system, and it reached the maximum value when catalpol was at 0.05 mg/L, while in 5~100 mg/L catalpol groups, the proliferation of OB was not increased.The ALP activity of OB in 0.05 mg/L catalpol group was higher than that in control group.The catalpal at 0.05 mg/L promoted the mRNA expression of ERβin OB in the co-culture system, but did not increase the mRNA expression of ERαas compared with control group. Catalpol at 0.05 mg/L obviously inhibited the bone resorption and the TRAP activity in OC.CONCLUSION: Catalpol stimulates the proliferation and activity of OB, inhibits the bone resorption and activity of OC, and increases the mRNA ex-pression of ERβin OB in the OB-OC co-culture system, suggesting that high mRNA expression of ERβmay be the regula-tory pathway of catalpol in response to bone metabolism.
5.The change of PAI-1 and TNF-α in old patients with metabolic syndrome and the effect of metformin
Manxiang DENG ; Huixia LIU ; Ji ZHANG ; Youbo YANG
Journal of Chinese Physician 2012;(12):1606-1608
Objective To explore the correlation of PAI-1 and TNF-α and the pathophysiology of the metabolic syndrome (MS) and coronary heart disease,and explore the role of metformin in the MS.Methods Sixty cases of old patients with the MS were chosen.These patients were divided into two groups at random.One group interfered with living style and metformin,the other group only interfered with living style.The activity of PAI-1 was detected by chromogenic substrate method,and the level of TNF-α was detected by ELISA assay.Results (1) The levels of PAI-1 and TNF-α in the MS patients [(0.95 ± 0.05) AU/ml,(24.81 ± 3.87)ng/ml] were significantly higher than in normal old people[(0.66 ± 0.10)AU/ml,(10.76 ±2.00) ng/ml] (P <0.001) ;(2)The levels of PAI-1 and TNF-α in the MS patients with CHD [(0.96 ± 0.05) AU/ml,(26.12 ± 2.83) ng/ml] were significantly higher than those in the patients without CHD [(0.94 ± 0.03) AU/ml,(23.71 ± 4.27) ng/ml] (P < 0.05) ;(3)The activity of PAI-1 and the level of TNF-α in the metformin group was decreased significantly [△ was (0.20 ± 0.17)AU/ml,(4.42 ± 0.85ng/ml),P <0.01],and metformin can improve the components of the MS.Conclusions The old patients with MS is prone to develop cardiac vascular disease.PAI-1 and TNF-α participate in pathophysiology of the MS and its complication.Metformin can inhibit the expression of PAI-1 and TNF-α to suppress the components of the MS,and block the complication of the MS.
6.The experience of the treatment of prostatic hyperplasia(BPH)with bladder calculus
Chinese Journal of Primary Medicine and Pharmacy 2012;19(2):211-212
Objective To evaluate the clinical effect and safety of ureteroscopic pneumatic lithotripsy combined with transurethral resection of prostate(TURP)for treatment of benign prostatic hyperplasia(BPH)with bladder calculus.Methods 68 patients of BPH with bladder calculus treated by ureteroscopic pneumatic lithotripsy combined with transurethral resection of prostate were reviewed.Results Satisfactory effects were achieved in all these 68 cases without serious complications.Conclusion Ureteroscopic pneumatic lithotripsy combined with TURP is a minimally invasive,safe and effective method in the treatment of BPH and bladder calculus.

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