1.AKT/MMP3 activation, not EMT pathway, participates in CCL20-induced invasion and migration of colon cancer SW480 cells
CHENG Xianshuo ; YANG Fang ; DONG Jian ; LI Yunfeng ; YANG Zhibin ; ZHANG Hongtao ; SHEN Tao ; LIU Ping ; YIN Zhengfeng ; LI Qiang
Chinese Journal of Cancer Biotherapy 2019;26(6):650-65
Objective: To investigate the molecular mechanism of chemokine CCL20/CCR6 in promoting invasion and migration of colon cancer SW480 cells. Methods: Colorectal cancer SW480 cells with high expression of CCR6 receptor were screened by immunochemistry (IHC). After co-culture with recombinant human CCL20, the invasion and migration of SW480 cells were detected by Transwell assay and Wound-Healing assay, respectively. Expressions of EMT markers, AKT signal protein and target protein MMP3 were detected by immunofluorescence (IF) and WB. AKT signaling pathway as the key mechanism was confirmed by MK2206 blocking assay. The expressions of CCL20 and MMP3 in colorectal cancer tissues as well as their correlation were analyzed by TCGAdatabase resources (https://portal.gdc.cancer.gov/). Results: CCL20 promoted the invasion and migration ability of SW480 cells significantly (all P <0.01), and this was induced by activation of AKT signaling and up-regulation of downstream target protein MMP3, instead of EMT. Blocking AKT signaling could significantly inhibit the invasion and migration of SW480 cells, and down-regulate MMP3 expression (P<0.05). TCGA platform data showed that the expressions of CCL20 and MMP3 in colorectal cancer tissues were significantly higher than those in normal mucosa tissues (P<0.05 or P<0.01), and an evidently positive correlation was found between CCL20 and MMP3 (r =0.051, P<0.01). Conclusion: The chemokine CCL20 promotes the invasion and migration of SW480 cells throughAKT/MMP3 signal axis, but not the EMT.
2.Chemical constituents of Fructus Gleditsiae Abnormalis
Lin MA ; Rongfei ZHANG ; Shule YU ; Zhengfeng WU ; Shouxun ZHAO ; Lei Wang ; Wencai YE ; Jian ZHANG ; Zhiqi YIN
Journal of China Pharmaceutical University 2015;46(2):188-193
Twelve compounds were isolated from the ethanol extract of Fructus Gleditsiae Abnormalis by macroporous resin, silica gel, Sephadex LH-20, MCI and ODS column chromatographies. Their structures were identified on the basis of physicochemical properties and spectral data as gleditsioside A(1), gleditsioside B(2), gleditsioside H(3), gleditsioside I(4), gleditsioside J(5), gleditsioside K(6), gleditsia saponins C′(7), tamarixetin-7-O-β-D-glucopyranoside(8), neohesperidin(9), chrysoeirol-7-O-neohesperidoside(10); syringaresinol- O-β-D-glucopyranoside(11), liriodendrin(12). Compounds 8-12 were firstly isolated from this genus.
3.Non-C21 steroids from the Rhizome of Cynanchum stauntonii
Shule YU ; Lin MA ; Zhengfeng WU ; Shouxun ZHAO ; Lei WANG ; Wencai YE ; Jian ZHANG ; Zhiqi YIN
Journal of China Pharmaceutical University 2015;46(4):426-430
Twelve compounds were isolated and purified from ethyl acetatefraction of Cynanchum stauntonii by silica gel, Sephadex LH-20 and ODS column chromatography. Their structures were identified by spectral techniques and physicochemical properties as syringaresinol(1), (-)-(7R, 7′R, 7″R, 8S, 8′S, 8″S)-4′, 4″-dihydroxy-3, 3′, 3″, 5-tetramethoxy-7, 9′ ∶7′, 9-diepoxy-4, 8″-oxy-8, 8′-sesquineolignan-7″, 9″-diol(2), prinsepiol(3), 4-hydroxyacetophenone(4), baishouwubenzophenone(5), 2, 4-dihydroxyacetophenone(6), benzoic acid(7), 1, 4-benzenediol(8), 6-O-[E]-sinapoyl-α-D-glucopyranoside(9a), 6-O-[E]-sinapoyl-β-D-glucopyranoside(9b), 1-O-methyl-α-D-cymadropyranoside(10), β-daucosterol(11), and β-sitosterol(12). Compounds 1-3, 5 and 7-11 were firstly isolated from this plant.
4.Advances on hypoglycemic activity and mechanism of triterpenoids
Kuiniu ZHU ; Zhengfeng WU ; Cuihua JIANG ; Lanjie CAO ; Jian ZHANG ; YIN YIN
Journal of China Pharmaceutical University 2015;46(6):764-770
The research of triterpenoids on hypoglycemic and anti-diabetic activities have made great progress. Findings indicated that triterpenoids could reduce blood glucose via different mechanisms, including increasing insulin secretion, enhancing insulin sensitivity, promoting glucose uptake by activation of AMP-activated protein kinase(AMPK), decreasing glycogenolysis and gluconeogenesis, and inhibiting protein tyrosine phosphates 1B(PTP1B), α-glycosidase, aldose reductase(AR)and dipeptidyl peptidase-4(DPP-4). This article reviews the hypoglycemic effects and mechanisms of triterpenoids, providing the reference for further research and development of triterpenoids.
6.Observation of intermediate-term curative effect of percutaneous laser disc decompression for the treatment of cervical spondylotic radiculopathy
Longxi REN ; Han GUO ; Tongtong ZHANG ; Zhengfeng HAN ; Jian YIN ; Xibin LIANG ; Xiaggfei ZHANG
Chinese Journal of Orthopaedics 2011;31(10):1044-1048
ObjectiveTo observe the 2-8 years' follow-up results of percutaneous laser disc decompression(PLDD) on cervical spondylotic radiculopathy,and to identify factors affecting the outcome of PLDD.MethodsEighty-seven patients with cervical spondylotic radiculopathy were treated consecutively by PLDD in our hospital from December 2002 to June 2009,who were followed up for 2-8 years.There were 32males and 26 females,with the mean age of 51.8 years (range,26-74).The results were evaluated according to the Japanese Orthopedic Association's score of cervical spondylotic radiculopathy(JOA 20 score) and numeric rating scales (NRS) after surgery.Two years after the operation,the excellent and good rate of JOA score of patients with different genders,ages and duration of each subgroup will be compared.ResultsThe excellent and good rate were 37.9%,51.7%,65.5%,81.0%,82.8%,80.9%,72.4% and 72.7% at 1,3,6months and 1,2,4,6,8 years after operation respectively.Significant difference was found between the time points of 6 months to eight years after operation and that of one month after operation.The NRS score of the pain symptoms at the final follow-up was significantly reduced(P<0.05).The excellent and good rate of patients whose course of disease (93.3%) was less than 18 months was significantly higher than that of those whose course of disease was over than 18 months(71.4%) two years after operation (P<0.05).Conclusion The intermediate-term curative effect of PLDD for the treatment of cervical spondylotic radiculopathy is reliable and stable,and the postoperative curative effect may be influenced by patients' duration of disease.
7.Extrahepatic recurrence after curative resection of hepatocellular cancer
Yanming ZHOU ; Jiamei YANG ; Bin LI ; Zhengfeng YIN ; Feng XU ; Tong KAN ; Feng XIE
Chinese Journal of General Surgery 2010;25(5):370-372
Objective To investigate the risk factors of extrahepatie recurrences after curative resection of primary hepatocellular carcinoma (HCC). Methods Clinicopathologic data of 238 curative resected cases of primary HCC were retrospectively reviewed for parameters affecting postoperative extrahepatic recurrences. Results During a median follow-up of 34 months (7 - 78 months), extrahepatic recurrences were observed in 32 out of 238 patients (13.4%). According to univariate analysis, the risk factors for extrahepatic recurrences were preoperative serum a-fetoprotein (AFP) level of > 1000 ng/ml,aspartate aminotransferase level of > 50 IU/L, presence of hepatic vein invasion, neighboring organ invasion, tumor satellites, and lack of tumor capsule formation. Preoperative serum AFP levels of > 1000 ng/ml, hepatic vein invasion, neighboring organ invasion proved to be independent risk factors by multivariate analysis. Conclusions Extrahepatic recurrences after curative resection of HCC was found to be related to preoperative serum AFP level of > 1000 ng/ml, hepatic vein invasion, and neighboring organ invasion.
8.Effects of troglitazone on the proliferation and differentiation of HepG2 cells
Yanming ZHOU ; Yinghao WEN ; Xiaoyan KANG ; Haihua QIAN ; Dianqi LI ; Jiamei YANG ; Zhengfeng YIN
Chinese Journal of Digestive Surgery 2008;7(3):200-202
Objective To examine the effects of a peroxisome proliferator-activated receptor γ ligand troglitazone on the proliferation and differentiation of HepG2 cells. Methods After the pretreatment of HepG2 cells with troglitazone, MTT and flow cytometry were used to analyze the proliferation and cell cycle of HepG2 cells, respectively. Immunocytochemistry, bromocresol green dye-binding method and chemiluminessence immunosorbent assay was used to determine E-cadherin, albumin and AFP, respectively. The expression of cyclin D1 and c-myc protein were detected by Western blot. Results Troglitazone inhibited the proliferation of HepG2 cells in a concentration-dependent manner and arrested HepG2 ceils at the G0>/G1> phase. After pretreated with troglitazone, HepG2 cells showed E-cadherin expression, a decreased expression of cyclin D1 and c-myc protein, a reduction of AFP level and a dramatic increase of albumin level. Conclusions Troglitazone inhibits proliferation and induces differentiation of HepG2 cells, the mechanism of which might be attributable to the down-regulation of cyclin D1 and c-myc expression.
9.The correlation between hepatitis virus infection and p53 expression in cholangiocarcinoma tissues
Yanming ZHOU ; Jiamei YANG ; Bin LI ; Zhengfeng YIN ; Feng XU ; Dianqi LI
Chinese Journal of General Surgery 2008;23(3):216-218
Objective To assess hepatitis B virus(HBV)and hepatitis C virus(HCV)infections in different anatomic location of cholangiocarcinoma(CC)and relationship with abnormal p53 expression.Methods A total of 411 CC samples including intrahepatic cholangiocarcinoma(ICC 312 cases);perihilar cholangiocarcinoma(PHC,73 cases)and distal cholangiocarcinoma(DC,26 cases)underwent serologic test for HBsAg and anti-HCV using microparticle enzyme immunoassay.Abnormal p53 expression was detected in formalin-fixed.paraffin-embedded CC tissues by immunohistochemistry.Results Seropositivity for HBsAg and anti-HCV were found in 48.4%(151/312)and 2.9%(9/312)of ICC cases,and in 10.9%(8/73)and zero of PHC,and in 7.7%(2/26)and zero of DC,respectively.Abnormal p53 expression was detected in 30.1%(94/312)of ICC cases.23.3%(17/73)of PHC cases and 19.2%(5/26)of DC cases.There was no correlation between seropositivity of HBsAg and anti-HCV and p53 overexpression among three groups of CC. Conclusions HBV but not HCV infection may be associated with the development of ICC.p53 abnormality may not play a significant role in HBV-associated carcinogenesis of ICC.
10.A strategy for searching antigenic regions in the SARS-CoV spike protein.
Yan REN ; Zhengfeng ZHOU ; Jinxiu LIU ; Liang LIN ; Shuting LI ; Hao WANG ; Ji XIA ; Zhe ZHAO ; Jie WEN ; Cuiqi ZHOU ; Jingqiang WANG ; Jianning YIN ; Ningzhi XU ; Siqi LIU
Genomics, Proteomics & Bioinformatics 2003;1(3):207-215
In the face of the worldwide threat of severe acute respiratory syndrome (SARS) to human life, some of the most urgent challenges are to develop fast and accurate analytical methods for early diagnosis of this disease as well as to create a safe anti-viral vaccine for prevention. To these ends, we investigated the antigenicity of the spike protein (S protein), a major structural protein in the SARS-coronavirus (SARS-CoV). Based upon the theoretical analysis for hydrophobicity of the S protein, 18 peptides were synthesized. Using Enzyme-Linked Immunosorbent Assay (ELISA), these peptides were screened in the sera from SARS patients. According to these results, two fragments of the S gene were amplified by PCR and cloned into pET-32a. Both S fragments were expressed in the BL-21 strain and further purified with an affinity chromatography. These recombinant S fragments were confirmed to have positive cross-reactions with SARS sera, either by Western blot or by ELISA. Our results demonstrated that the potential epitope regions were located at Codons 469-882 in the S protein, and one epitope site was located at Codons 599-620. Identification of antigenic regions in the SARS-CoV S protein may be important for the functional studies of this virus or the development of clinical diagnosis.
Antigens, Viral
;
immunology
;
Chromatography, High Pressure Liquid
;
Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
;
Enzyme-Linked Immunosorbent Assay
;
Genetic Vectors
;
Humans
;
Mass Spectrometry
;
Membrane Glycoproteins
;
genetics
;
immunology
;
metabolism
;
Molecular Weight
;
Peptide Fragments
;
chemistry
;
Recombinant Proteins
;
genetics
;
immunology
;
SARS Virus
;
genetics
;
immunology
;
metabolism
;
Spike Glycoprotein, Coronavirus
;
Viral Envelope Proteins
;
genetics
;
immunology
;
metabolism

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