1.miR-18a-5p Regulates Colorectal Cancer Proliferation and Progression by Targeting RORA
Yifeng CHEN ; Shuai WANG ; Mingming CHAI ; Di ZHANG ; Chunxia WANG ; Lixia ZHAO ; Honglai ZHANG ; Xiongfei YANG ; Weisheng ZHANG ; Tao WANG
Cancer Research on Prevention and Treatment 2024;51(8):667-677
Objective To investigate the mechanism and clinical significance of miR-18a-5p and retinoid acid receptor-related orphan receptor-α(RORA)in the proliferation and progression of colorectal cancer(CRC)cells.Methods The expressions of miR-18a-5p and RORA in CRC cells and tissues were detected via qRT-PCR,FISH,and IHC.Cell proliferation capability was detected through EdU and CFSE assay,cell apoptosis by flow cytometry assay,and cell migration and invasion abilities by cell scratch and Transwell invasion assays,respectively.The targeted regulation of miR-18a-5p on RORA was further verified via dual-luciferase reporter assay,cell function rescue test,RT-PCR,and Western blot assay.Finally,bioinformatics was used to explore the molecular mechanism of miR-18a-5p promoting malignant proliferation,invasion,and progression of CRC via regulating RORA.Results miR-18a-5p exhibited a high expression in CRC tissues and cells(P<0.05)and promoted the proliferation,migration,and invasion of CRC cells(P<0.05).In addition,RORA served as the target gene of miR-18a-5p,and its overexpression effectively reduced the promoting function of miR-18a-5p in the malignant biological phenotype of CRC cells(P<0.05).The expression of RORA in CRC tissues showed a significantly positively correlation with the infiltration of CD8+T cells and the expression of its surface marker protein CD8A.Conclusion The targeted regulation of RORA by miR-18a-5p promotes the proliferation and progression of CRC.The miR-18a-5p/RORA regulatory pathway possibly contributes to the immune microenvironment of CRC,which can be a potential therapeutic target for CRC.
2.Effects and mechanism of galangin on hepatocyte apoptosis in rats with obstructive jaundice
Lin ZHAO ; Zuowu XI ; Shasha XU ; Xubiao PAN ; Weisheng GUO
China Pharmacy 2024;35(18):2246-2251
OBJECTIVE To investigate the effects and mechanism of galangin (GAL) on hepatocyte apoptosis in rats with obstructive jaundice (OJ) based on the Janus kinase 2 (JAK2)/signal transduction and activator of transcription 3 (STAT3) signaling pathway. METHODS Taking male SD rats as the object, the OJ model was established by double ligation of common bile duct, and 48 rats with successful modeling were randomly separated into OJ model group (model group), low-dose GAL group (GAL-L group), high-dose GAL group (GAL-H group) and high-dose GAL+JAK2 activator colivelin group (GAL-H+ colivelin group), with 12 rats in each group; another 12 SD rats with laparotomy/abdominal closure without ligation were selected as sham operation group (sham group). Each administration group was given relevant medicine intragastrically and/or intraperitoneally, once a day, for 7 consecutive days. After the last medication, the morphology of liver tissue in rats was observed; the serum levels of total bilirubin (TBIL), direct bilirubin (DBIL), alanine transaminase (ALT) and γ -glutamyltransferase (GGT), as well as the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in mail:guoweishengjcy@126.com liver tissue were detected. The apoptotic rate of liver tissue cells, the expression levels of signaling pathway-related proteins (phosphorylated JAK2, JAK2, phosphorylated STAT3, STAT3) and apoptosis-related proteins [B cell lymphoma 2 (Bcl-2), Bcl-2 related X protein (Bax)] were determined. RESULTS Compared with sham group, congestion of liver sinusoids, damage to liver lobules, disordered arrangement and swollen morphology of liver cells, the disappearance of nucleoli, and significant infiltration of inflammatory cells and fibrous tissue proliferation were observed in model group; the serum levels of TBIL, DBIL, ALT and GGT, the level of MDA in liver tissue, the apoptosis rate of liver cells, the protein expression of Bax, and the protein phosphorylation levels of JAK2 and STAT3 in liver tissue of model group were increased significantly (P<0.05); the level of SOD and the protein expression of Bcl-2 in liver tissue were decreased significantly (P<0.05). Compared with the model group, the pathological injuries of liver tissue were relieved in GAL-L group and GAL-H group, all quantitative indicators had significantly improved, and the effect of GAL-H group was more significant (P< 0.05). Colivelin could significantly reverse the improvement effects of GAL on liver injury and related indicators of OJ rats (P< 0.05). CONCLUSIONS GAL may inhibit liver cell apoptosis in OJ rats, improve liver function and alleviate oxidant stress, the mechanism of which may be associated with inhibiting JAK2/STAT3 signaling pathway.
3.2,3-Seco and 3-nor guaianolides fromAchillea alpina with antidiabetic activity.
Guimin XUE ; Chenguang ZHAO ; Jinfeng XUE ; Jiangjing DUAN ; Hao PAN ; Xuan ZHAO ; Zhikang YANG ; Hui CHEN ; Yanjun SUN ; Weisheng FENG
Chinese Journal of Natural Medicines (English Ed.) 2023;21(8):610-618
In this study, we presented the isolation and characterization of eight novel seco-guaianolide sesquiterpenoids (1-8) and two known guaianolide derivatives (9 and 10), from the aerial part of Achillea alpina L.. Compounds 1-3 were identified as guaianolides bearing an oxygen insertion at the 2, 3 position, while compounds 4-8 belonged to a group of special 3-nor guaianolide sesquiterpenoids. The structural elucidation of 1-8, including their absolute configurations, were accomplished by a combination of spectroscopic data analysis and quantum electronic circular dichroism (ECD) calculations. To evaluate the potential antidiabetic activity of compounds 1-10, we investigated their effects on glucose consumption in palmitic acid (PA)-mediated HepG2-insulin resistance (IR) cells. Among the tested compounds, compound 7 demonstrated the most pronounced ability to reverse IR. Moreover, a mechanistic investigation revealed that compound 7 exerted its antidiabetic effect by reducing the production of the pro-inflammatory cytokine IL-1β, which was achieved through the suppression of the NLRP3 pathway.
Humans
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Hypoglycemic Agents/pharmacology*
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Circular Dichroism
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Cytokines
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Glucose
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Hep G2 Cells
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Insulin Resistance
4.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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5.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.
6.The relationship between the neuromyelitis optica astrocyte damage and the patients with functional defect
Xiaoling CHEN ; Yin LEI ; Yulan TANG ; Chengcheng YANG ; Weisheng ZHAO
The Journal of Practical Medicine 2016;32(11):1767-1771
Objective To explore the relationship between the damage of neuromyelitis optica (NMO) astrocytes (AS) and the onset of NMO, and investigate the relevance of AS damage with the severity of the patients with functional defect. Methods The levels of aquaprin4-antibody (AQP4 -Ab), glial fibrillary acidic protein (GFAP), apolipoprotein(ApoE),interleukins-6(IL-6), interleukins-10(IL-10), tumor necrosis factor-α(TNF-α) in cerebrospinal fluid (CSF ) and serum of 30 acute NMO patients were tested by means of ELISA. The results were later compared with control group. And analysis of the relevance of the various index of the levels in CSF with the CSF AQP4-Ab level, the acute phase expanded disability status scale(EDSS) score of the NMO group were made. Results (1)The NMO group in CSF and serum AQP4-Ab, GFAP, IL-6 levels were higher than the control group (P < 0.05), and ApoE, IL-10 levels were lower than the control group (P < 0.05). (2)The CSF GFAP, ApoE, IL-6 in NMO group is higher than the serum (P < 0.05), and CSF AQP4-Ab, IL-10 levels were lower than the serum ( P < 0 . 05 ) . ( 3 ) The CSF GFAP , IL-6 levels and the CSF AQP4-Ab level were positively correlated (r=0.749, r=0.526, P<0.05), and the CSF ApoE, IL-10 levels were negatively correlated with CSF AQP4-Ab level(r = -0.571, r = -0.676, P < 0.05). (4)The CSF AQP4-Ab, GFAP, IL-6 levels and the acute phase EDSS score were positively correlated (P < 0.05), the CSF ApoE, IL-10 levels were negatively correlated with the acute phase EDSS score (P < 0.05). Conclusion The AS damage exists in the NMO and the damage severity may correlate with patient function defect. AQP4-Ab, GFAP, IL-6 may play important roles in the onset of NMO and the disease aggravating. The decrease of the ApoE and IL-10 may exacerbate NMO damage.
7.Cloning, Sequence Analysis and Prokaryotic Expression ofGGPS Gene fromLepidium apetalum
Ligang MA ; Le ZHAO ; Yingchao LI ; Weisheng FENG ; Haixue KUANG ; Xiaoke ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):485-491
This study was aimed to clone the GGPS (geranylgeranyl pyrophosphate synthase) gene from Lepidium apetalum, to analyze its sequence, and to express the protein in E.coli expression system. Specific PCR cloning primers were designed for GGPS gene from Lepidium apetalum according to the full-length sequence from a previous transcriptome sequencing project. PCR amplification was performed with this primer pair on a leaf cDNA template. TA cloning, sequencing and sequence analysis were performed.GGPS gene from Lepidium apetalum was expressed in the E.coli expression system. The results showed that the full-lengthGGPS cDNA from Lepidium apetalum was 1 146 bp coding a protein of 381 amino acids. The LaGGPS protein had an isoprenoid synthase domain. According to a phylogenetic tree constructed with multiple alignment of GGPS protein sequences from various plant species, GGPS protein from Lepidium apetalum was the closest to Arabidopsis thaliana and Sinapis alba. The prokaryotic expression vectorpET-32a-LaGGPS was also constructed successfully. The protein was expressed in E.coli BL21 strain. It was concluded that the cloning and prokaryotic expression of LaGGPS gene provided a foundation for a follow-up research of its function with protein purification and activity analysis.
8.Research on Fraction-splitting Technology of Seeds ofDescurainia Sophia
Minghui ZHANG ; Yanli ZHANG ; Wei ZHAO ; Xiaolan WANG ; Weisheng FENG ; Haixue KUANG ; Xiaoke ZHENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(3):476-484
This study was aimed to establish the method for fraction-splitting of seeds of Descurainia sophia, in order to study the stability and unoverlapping property of fractions of seeds of D. sophia. The fraction-splitting of seeds of D. sophia were obtained through the combination of various methods including decoction, distillation, extraction, ethanol precipitation and gradient elution of macroporous adsorption resin. HPLC and chemometrics software were used to analyze the stability and unoverlapping property of the fractions of D. sophia. The results showed that the chemical components from seeds of D. sophia was divided into six fractions. HPLC data and chemometrics analysis showed good stability of technology. The fraction-splitting of seeds of D. sophia was unoverlapping. It complied with the research model of chemical constituents of seeds of D. sophia which can be split and combinatorial. It was concluded that the established method for splitting fractions of seeds of D. sophia had good stability and repeatability. Each splitting fraction uncrossed others.
9.Hepatitis B virus X protein induces epithelial-mesenchymal transition through c-Src activation in liver cancer cells
Shizhong YANG ; Zhiyuan ZHAO ; Weisheng YUAN ; Gen CHEN ; Yujun ZHANG ; Aiqun ZHANG ; Jiahong DONG
Chinese Journal of Hepatobiliary Surgery 2015;21(3):200-203
Objective To examine the role of c-Src activation in hepatitis B virus X (HBx) protein induced epithelial-mesenchymal transition (EMT) in liver cancer.Methods SMMC-7721 liver cancer cells were transfected with HBx gene to induce EMT and the activated c-Src expression was evaluated by Western blot.Both the morphological changes and the epithelial and mesenchymal markers expression (real-time PCR,western blot and immunocytochemistry) of HBx-transfected SMMC-7721 cell treated by c-Src kinase inhibitor PP2 and negative control PP3 were observed and compared,respectively.Results The activated c-Src expression in HBx gene transfected SMMC-7721 cells was significantly increased compared to that in mock transfected cells,c-Src kinase inhibitor PP2 could enable the HBx-transfected SMMC-7721 cells to transmit from spindle-like shape to original epithelial morphology.Western blot and immunocytochemistry confirmed that the expression of epithelial markers and mesenchymal markers almost returned to the levels of parental cells,indicating the mesenchymal-epithelial transition.Conclusions c-Src activation plays a key role in the process of EMT induced by HBx protein in SMMC-7721 cells.
10.Investigation on compliance of forbidden depasturing livestock on marshland with Oncomelania snails in schistosomiasis endemic areas
Chunli CAO ; Jing WANG ; Ziping BAO ; Hongqing ZHU ; Shunxiang CAI ; Yiyi LI ; Dong LI ; Jiachang HE ; Leping SUN ; Xianhong MENG ; Bo ZHONG ; Xiguang FENG ; Zhengming SU ; Jun LI ; Xiaonan GU ; Hao WANG ; Weiwei RU ; Weisheng JIANG ; Shizhu LI ; Qiang WANG ; Xiaonong ZHOU ; Jiagang GUO ; Gengming ZHAO
Chinese Journal of Schistosomiasis Control 2009;21(6):482-485
Objective To study the compliance of forbidden depasturing livestock on the marshland with Oncomelania snails in schistosomiasis endemic areas. Methods According to 3 levels of human infection rates as > 10% ,5%-10% and <5% , 2 204 residents selected randomly from the schistosomiasis endemic villages were sampled with the stratified cluster sampling method in Hunan, Hubei, Jiangxi, Anhui, Jiangsu, Sichuan and Yunnan provinces, and investigated by questionnaire. The contents of the questionnaire included the recognition and implementation of forbidden depasturing livestock on marshland with Oncomelania snails and breeding livestock in bam. Results A total of 78.4% residents agreed forbidden depasturing livestock on marshland with snails, but 3. 7% residents disagreed it. A total of 83. 9% residents considered the relationship between breeding livestock in bam and schistosomiasis control, but 3. 1% residents thought that it was no relationship. The main reasons of depasturing livestock on marshland with Oncomelania snails were the high cost of breeding livestock in bam (36. 2% ) , unaccustomed (26.4% ) and no room for breeding livestock in bam (25.4% ). Conclusion Forbidden depasturing livestock on the marshland with Oncomelania snails should be strengthened according to the local economic, nature environment, agriculture, residents'culture degree and agriculture habit.

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