1.Rehmanniae Radix Praeparata Improves Neurological Function of Ischemic Stroke Rats by Inhibiting Autophagy and Ferroptosis
Saifei LI ; Peipei YUAN ; Yaxin WEI ; Liyuan GAO ; Panying LI ; Yuan RUAN ; Yi CHEN ; Yang FU ; Xiaoke ZHENG ; Weisheng FENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):26-33
ObjectiveTo investigate the effect of Rehmanniae Radix Praeparata on neurological function injury in ischemic stroke rats and explore its mechanism. MethodMale SD rats were randomized into sham operation, model, low- and high -dose (3.5 g·kg-1 and 7 g·kg-1) Rehmannia Radix Praeparata, and nimodipine (0.01 g·kg-1) groups. The rat model of middle cerebral artery occlusion (MCAO) was established with the modified suture occlusion method. Zea-Longa 5-point scoring was employed to evaluate the neurological function of rats. The cerebral infarction volume was detected by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Hematoxylin-eosin staining and Nissl staining were employed to observe the morphology and damage of the brain tissue. Meanwhile, the serum levels of lactate dehydrogenase (LDH), oxidative stress-related indicators superoxide dismutase (SOD), glutathione peroxidase 4 (GPX4), and malondialdehyde (MDA), and the iron (Fe) content in the brain tissue were determined. To explore the mechanism of Rehmanniae Radix Preparata in mitigating the neurological damage in ischemic stroke rats, Western blotting was employed to determine the expression levels of proteins in the ischemic brain tissue. The autophagy-associated proteins included autophagy effector (beclin-1), microtubule-associated protein light chain 3 (LC3B), and ubiquitin-binding protein p62 (p62). The ferroptosis-associated proteins included transferrin (TF), transferrin receptor 1 (TFR1), ferritin heavy chain 1 (FTH1), and ferropotin (FPN1). The neurological function injury-associated proteins included brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB). ResultCompared with the sham operation group, the model group showed increased neurological function score, cerebral infarction volume, and appearance of nuclear pyknosis and vacuole of cells in the cerebral cortex. In addition, the model group presented elevated levels of LDH, MDA, and Fe (P<0.01) and lowered levels of SOD and GPX4 (P<0.01). Compared with the model group, Rehmanniae Radix Praeparata decreased the content of LDH, MDA, and Fe (P<0.05, P<0.01) and elevated the levels of SOD and GPX4 (P<0.05, P<0.01). Compared with the sham operation group, the modeling promoted the expression of beclin-1,LC3B Ⅱ/Ⅰ, TF, and TFR1 and inhibited the expression of p62, FTH1, FPN1, BDNF, and TrkB (P<0.01). The expression levels of these proteins were recovered after the treatment with Rehmanniae Radix Praeparata. ConclusionRehmanniae Radix Praeparata may inhibit ferroptosis and improve the neurological function in ischemic stroke rats by down-regulating the autophagy level in the brain tissue.
2.Retrospective analysis of 350 cases with dissection of lymph nodes posterior to right recurrent laryngeal nerve in endoscopic thyroidectomy through gasless axillary posterior approach
Zhicheng ZHANG ; Tingting LI ; Shitong YU ; Junna GE ; Zhigang WEI ; Baihui SUN ; Weisheng CHEN ; Jie TAN ; Shangtong LEI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(1):21-26
Objective:To evaluated the safety and feasibility of dissection of lymph nodes posterior to right recurrent laryngeal nerve (ⅥB compartment) in endoscopic thyroidectomy through gasless axillary posterior approach.Methods:A total of 350 cases with right lobe papillary thyroid carcinoma (PTC) who underwent endoscopic lobectomy, isthmusectomy and central compartment neck dissection via gasless axillary posterior approach based at the Department of General Surgery, Nanfang Hospital, Southern Medical University from June 2020 to December 2022 were retrospectively analyzed. Summarize the clinical, pathological characteristics, and postoperative complications of the patients. SPSS 25.0 was used for statistical analysis of the data.Results:All 350 patients underwent endoscopic surgery successfully, with no conversion to open surgery. There were 303 females and 47 males, with an average age of (36.3±9.2) years. Of those, 287 patients were in pT1a stage, 62 in pT1b stage, and one patient in pT2 stage. There was no T3 or T4 stage patient. The mean numbers of yielded lymph nodes in right central compartment and ⅥB compartment were 8.11±4.65 (range, 1-31) and 2.62±1.86 (range, 1-12), respectively. ⅥB compartment metastasis was detected in 52 (14.86%) of 350 patients. The incidence of transient recurrent laryngeal nerve injury was 0.86%(3/350). Postoperative hematoma occurred in three patients (0.86%).Conclusion:The dissection of ⅥB compartment in endoscopic thyroidectomy through gasless axillary posterior approach is safe and feasible in selected PTC patients
3.Study on comprehensive quality evaluation of Sargentodoxae Caulis from different habitats
Weisheng LYU ; Cuijie WEI ; Zhenyu LI ; Hui LIANG ; Binbin SHEN ; Xiangdong CHEN ; Congyou DENG ; Xiaozhou JIA
International Journal of Traditional Chinese Medicine 2023;45(8):1004-1010
Objective:To comprehensively evaluated the quality of Sargentodoxae Caulis from different habitats with a combination of indexes and characteristic chromatogram method from Chinese Pharmcopoeia (Edition 2020). Methods:The contents of water content, total ash, ethanolic extract, sulfur dioxide residue, heavy metals and harmful elements, total phenols, chlorogenic acid, salidroside and characteristic chromatogram of 17 batches of Sargentodoxae Caulis were determined. The quality of Sargentodoxae Caulis was comprehensively evaluated by combining chemical pattern recognition method. Results:The water content, total ash content, extracts, and content determination of 17 batches of Sargentodoxae Caulis from different habitats complyed with the provisions of the Chinese Pharmcopoeia (Edition 2020). There were differences in the contents of extracts, chlorogenic acid, and salidroside, among which the content of Anhui origin was higher. A total of 8 common peaks were identified from the 17 batches samples. Conclusion:Comprehensive evaluation of multiple indicators can demonstrate the quality of Sargentodoxae Caulis more correctly, and shows that the quality of Sargentodoxae Caulis from different habitats is different. The quality of Sargentodoxae Caulis from Anhui is better than that from other habitats.
4.Quality study of Perillae caulis formula granules based on standard decotion
Jie YANG ; Weisheng LYU ; Fang CHEN ; Liping DENG ; Ruoyun SHI ; Zhiwen DUAN ; Minyou HE ; Mei WEI ; Zhenyu LI
International Journal of Traditional Chinese Medicine 2022;44(10):1144-1149
Objective:To establish the quality evaluation method of Perillae caulis formula granules based on the three kind of quality indexes of standard decoction. Methods:Eighteen batches of Perillae caulis were collected from different habitats according to different technical requirements, eighteen batches of standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The content of Caffeic acid and Rosmarinic acid were determined and calculated by Ultra High Performance Liquid Chromatography (UPLC). Then the fingerprints of standard decoction of and formula granules of Perillae caulis were established by UPLC . The similarity values of fingerprints between formula granules and standard decoction were calculated. Results:The average paste-forming rate of standard decoction was (7.16±1.97)%. The paste-forming rates of three batches of formula granules were 5.52%, 5.25% and 5.34%, respectively. The average content of Caffeic acid and Rosmarinic acid in standard decoction was (12.06±3.37)mg/g. The contents of three batches of formula granules were 5.52, 5.82, 5.77 mg/g, respectively. Seven common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, three of which were identified as Caffeic acid, N-Feruloyl Octopus amine and Rosmarinic acid by comparison of reference substance. The fingerprints similarity of Perillae caulis dispensing granules and standard decoction were 1.000, 0.995 and 0.997, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method can provide basis for the establishment of quality standard of Perillae caulis dispensing granules.
5.Establishment of the fingerprint of Cnidium monnieri and a method for the content determination of 4 kinds of coumarins
Xiaojun PAN ; Weisheng LYU ; Wenhui YANG ; Yuqin LUO ; Yueyi LIANG ; Mei WEI ; Dongmei SUN ; Xiangdong CHEN ; Wenjie HUO ; Zhenyu LI
China Pharmacy 2022;33(2):185-190
OBJECTIVE To establi sh the fingerprint of Cnidium monnieri and a method for the content determination of 4 kinds of coumarins. METHODS Ultra-high performance liquid chromatography (UPLC) method was adopted to establish the fingerprints of 21 batches of C. monnieri ; their similarities were evaluated with Similarity Evaluation System of TCM Chromatographic Fingerprint (2012 edition);common peaks were identification by comparison with reference substance. Using 10 common peak areas as variables ,cluster analysis was performed for 21 batches of C. monnieri by the method of between groups. The relative correction factors of xanthotoxin ,bergapten and imperatorin were calculated by the same UPLC method with osthole as the internal reference. The contents of them were calculated by quantitative analysis of multi-components by single marker (QAMS),and compared with the results of external standard method. RESULTS Totally 10 common peaks were identified in the fingerprints of 21 batches of C. monnieri ;the similarities ranged from 0.997 to 1.000. Peak 4 was identified as xanthotoxin ,peak 8 as bergapten ,peak 9 as imperatorin and peak 10 as osthole. A total of 21 batches of samples were divided into 3 categories,of which S 7 was clustered into one category ,S14 was clustered into one category ,and the other 19 batches were clustered into one category. The relative deviations of the contents of xanthotoxin ,bergapten and imperatorin determined by QAMS and external standard method were in the range of 0.88% -1.07% ,2.22% -2.29% ,0.67% -2.93% ,respectively. CONCLUSIONS UPLC fingerprint of C. monnieri is successfully established ,and QAMS method for content determination of 4 coumarins is also established.
6.Optimization of Extraction Technology of Total Triterpenes from the Leaves of Cornus officinalis by Box-Behnken Design-response Surface Methodology
Denghui ZHU ; Jingke ZHANG ; Meng LI ; Jingya SHI ; Juanjuan LIU ; Junjun WEI ; Xiaoke ZHENG ; Weisheng FENG
China Pharmacy 2021;32(1):46-50
OBJECTIVE:To optimize the extraction technology for total triterpenes from the leaves of Cornus officinalis . METHODS:Based on the full swelling of the leaves of C. officinalis ,total triterpenes was extracted with heating reflux method. The effects of ethanol concentration ,liquid-solid ratio ,extraction time and extraction times on the contents of total triterpenes from the leaves of C. officinalis were investigated by single factor test. Using oleanolic acid as control ,the contents of total triterpenes were detected by UV spectrometry. On the basis of single factor test ,fixing the times of extraction a s 3 times,taking the contents of total triterpenes as response value ,using ethanol volume fraction ,solid-liquid ratio and extraction time as factors , Box-Behnken design-response methodology was used to optimize the extraction technology of total triterpenes from the leaves of C. officinalis,and the optimized extraction technology was validated. RESULTS :The optimal extraction technology of total triterpenes from the leaves of C. officinalis were as follows as ethanol concentration of 73%,liquid-to-material ratio of 38 ∶ 1(mL/g), extraction time of 60 min. Results of 3 validation tests showed that the contents of total triterpenes from the leaves of C. officinalis were 6.92%,6.91%,6.84%;the average content was 6.89%(RSD=0.63%),relative error of which with the predicted value (7.28%)was 5.36%. CONCLUSIONS :The optimized technology is stable and reliable ,and can be used for the extraction of total triterpenes from leaves of C. officinalis .
7.Expert Consensus for Thermal Ablation of Pulmonary Subsolid Nodules (2021 Edition).
Xin YE ; Weijun FAN ; Zhongmin WANG ; Junjie WANG ; Hui WANG ; Jun WANG ; Chuntang WANG ; Lizhi NIU ; Yong FANG ; Shanzhi GU ; Hui TIAN ; Baodong LIU ; Lou ZHONG ; Yiping ZHUANG ; Jiachang CHI ; Xichao SUN ; Nuo YANG ; Zhigang WEI ; Xiao LI ; Xiaoguang LI ; Yuliang LI ; Chunhai LI ; Yan LI ; Xia YANG ; Wuwei YANG ; Po YANG ; Zhengqiang YANG ; Yueyong XIAO ; Xiaoming SONG ; Kaixian ZHANG ; Shilin CHEN ; Weisheng CHEN ; Zhengyu LIN ; Dianjie LIN ; Zhiqiang MENG ; Xiaojing ZHAO ; Kaiwen HU ; Chen LIU ; Cheng LIU ; Chundong GU ; Dong XU ; Yong HUANG ; Guanghui HUANG ; Zhongmin PENG ; Liang DONG ; Lei JIANG ; Yue HAN ; Qingshi ZENG ; Yong JIN ; Guangyan LEI ; Bo ZHAI ; Hailiang LI ; Jie PAN
Chinese Journal of Lung Cancer 2021;24(5):305-322
"The Expert Group on Tumor Ablation Therapy of Chinese Medical Doctor Association, The Tumor Ablation Committee of Chinese College of Interventionalists, The Society of Tumor Ablation Therapy of Chinese Anti-Cancer Association and The Ablation Expert Committee of the Chinese Society of Clinical Oncology" have organized multidisciplinary experts to formulate the consensus for thermal ablation of pulmonary subsolid nodules or ground-glass nodule (GGN). The expert consensus reviews current literatures and provides clinical practices for thermal ablation of GGN. The main contents include: (1) clinical evaluation of GGN, (2) procedures, indications, contraindications, outcomes evaluation and related complications of thermal ablation for GGN and (3) future development directions.
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8.Variations of Chemical Components in Inula japonica by UPLC-MS/MS before and after Honey-frying
Weisheng LYU ; Cuijie WEI ; Xiaojun PAN ; Wenhui YANG ; Minyou HE ; Xiangdong CHEN ; Dongmei SUN ; Mei WEI ; Zhenyu LI
China Pharmacy 2021;32(20):2478-2484
OBJECTIVE:To compare the c hemical components differences of Inula japonica before and after honey-frying. METHODS:UPLC-MS/MS method was adopted. The determination was performed on Waters ACQUITY UPLC BEH C 18 column with mobile phase consisted of 0.1% formic acid-acetonitrile (gradient elution )at the flow rate of 0.3 mL/min. The column temperature was set at 30 ℃,and sample size was 5 µL. The electrospray ion source was scanned by positive ion mode. The first order mass spectrometry scanning range was m/z 70-1 050,the second order mass spectrometry scanning range was m/z 50-1 050, and the normalized collision energy was 40,60 eV ;mass spectrum type was the peak figure ,the flow rate of sheath gas was 35 arb,the auxiliary airflow speed was 10 arb,the spray voltage was 3.80 kV,the S-lens voltage was 50 V,the heating temperature was 350 ℃,and the capillary temperature was 350 ℃. The components were identified by Qual Browser 4.1.39.1 software, referring to the online high-resolution database mzCloud and local database OTCML of high-resolution mass spectrometry of TCM , and combined with relevant literature. The principal component analysis (PCA)and orthogonal partial least squared-discriminant analysis(OPLS-DA)of I. japonica before and after honey-fried were performed by using SIMCA 14.1 statistical software ,and variable importance projection (VIP)value greater than 1 was used as the standard to screen the differential components before and after honey-frying. RESULTS :A total of 29 common chemical components were identified from I. japonica and honey-fried I. japonica,including 5 phenolic acids as 1-caffeoylquinic acid ,chlorogenic acid and 3,5-dicaffeoylquinic acid ,12 flavonoids as quercetin,luteolin and evamectin ,as well as 12 sesquiterpene lactones as 1-O-acetylinula diester ,inula bicolor lactone B and 1-O-acetyl-6-O-isobutyryl inulin. The results of PCA showed that I. japonica and honey-fried I. japonica were located on both sides of the score diagram respectively. The results of OPLS-DA showed that the VIP values of 7 components were greater than 1,which were peak 19(britanin),peak 6(quercetagitrin),peak 1(1-caffeoylquinic acid ),peak 21(vitexicarpin),peak 20(tomentosin), peak 13(spinacetin)and peak 3(daphnetin). CONCLUSIONS :After honey-fried ,the content of chemical components of I. japonica changed and decreased to a certain extent. Britanin ,quercetagitrin,1-caffeoylquinic acid ,tomentosin,vitexicarpin, spinacetin and daphnetin may be the differential components of I. japonica and honey-fried I. japonica .
9.Quality Evaluation of Polygonum cuspidatum by UPLC Fingerprint and QAMS
Hui LIANG ; Xiaojun PAN ; Wenhui YANG ; Weisheng LYU ; Yuqin LUO ; Jiaxuan WU ; Mei WEI ; Xueren CHENG ; Xiaoyong LAN ; Zhenyu LI
China Pharmacy 2021;32(15):1842-1848
OBJECTIVE:To establish the UPLC fingerprint of Polygonum cuspidatum ,and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS :The determination was performed on Waters BEH C 18 column(100 mm×2.1 mm,1.7 μm)with mobile phase consisted of acetonitrile- 0.2% formic acid (gradient elution )at flow rate of 0.4 mL/min. The column temperature was 40 ℃,and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation,cluster analysis and orthogonal partial least square discriminant analysis (OPLS-DA). Using polydatin as internal standard ,relative calibration factors of resveratrol ,emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS:UPLC fingerprints of 15 batches of P. cuspidatum were established ,and 12 common peaks were confirmed. Five components were identified ,i.e. polydatin ,resveratrol,emodin-8-O-β-D-glucoside,emodin,emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ,15 batches of P. cuspidatum were classified into 4 categories,showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7>emodin-8-O-β-D-glucoside>resveratrol>peak 8>polydatin>peak 1> peak 10. The relative deviation among the contents of resveratrol ,emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0%,indicating that there was no significant difference between the two methods. CONCLUSIONS :UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ;the quality of P. cuspidatum produced in Chongqing and Anhui province is better.
10.miR-375 affects the proliferation and invasion of hepatoma cells via targetingYAP
ZHANG Xiaolu ; DU Meihong ; CHEN Hongyue ; ZHANG Jiangang ; WEI Guangya ; ZHAO Lin ; GUO Weisheng
Chinese Journal of Cancer Biotherapy 2019;26(1):79-84
Objective: To investigate the mechanisms of miR-375 affecting the proliferation and invasion of hepatoma cells via targeting YAP (Yes-associated protein). Methods: The cancerous tissues and corresponding para-cancerous tissues of 70 patients with hepatocellular carcinoma (HCC) who underwent surgical resection at the Second Affiliated Hospital of Henan University of Traditional Chinese Medicine from January 2015 to December 2016, as well as the hepatoma cell lines SMMC-7721, Hb611, HepG2 and BEL-7405 were collected for this study. qPCR method was used to detect the expression level of miR-375 in collected HCC tissues and different hepatoma cell lines; Dual luciferase reporter gene assay was used to verify the interaction between miR-375 and YAP; The relationship between miR-375 and clinicopathological features of HCC patients was also analyzed; MTT assay was used to detect the effect of miR375 on the proliferation of hepatoma cells; Transwell invasion assay was used to detect the invasive ability of hepatoma cells after inhibiting the expression of miR-375; Western blotting was used to detect the expression of YAP in HepG2 cells. The nude mouse model of subcutaneously transplanted xenograft was established, and the tumor volume and mass of transplanted hepatoma cells were detected after inhibiting the expression of miR-375. The expression of YAP in xenograft of nude mice was detected by immunohistochemistry and Western blotting. Results: The expression of miR-375 and YAP in HCC tissues was significantly higher than that in para-cancerous tissues (all P<0.05). The expression of miR-375 in HepG2 cells was the highest (P<0.05). miR-375 could specifically bind to the 3' UTR of YAP and regulate the expression activity of YAP. After inhibiting the expression of miR-375, the proliferation and invasion abilities of HepG2 cells were reduced (all P<0.05); The tumor volume and mass of transplanted xenografts were significantly reduced (both P<0.05); The expression of YAP protein in the transplanted xenografts was down-regulated (P<0.05). Conclusion: miR-375 plays an important role in the occurrence and development of liver cancer, and can influence the malignant biological behaviors of hepatoma cells by targeting and regulating the expression ofYAP.

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