1.Methyl ferulic acid ameliorates ethanol-induced L02 cell steatosis through microRNA-378b-mediated CaMKK2-AMPK pathway.
Ping HUANG ; Xing CHEN ; Rong-Hua MENG ; Jun LU ; Yan ZHANG ; Li LI ; Yong-Wen LI
China Journal of Chinese Materia Medica 2023;48(1):193-201
Alcoholic liver disease(ALD), with its increasing morbidity and mortality, has seriously and extensively affected the health of people worldwide. Methyl ferulic acid(MFA) has been proven to significantly inhibit alcohol-induced lipid production in L02 cells through the AMP-activated protein kinase(AMPK) pathway, but its in-depth mechanism remains unclear. This study aimed to further clarify the mechanism of MFA in improving lipid accumulation in L02 cells through the microRNA-378b(miR-378b)-mediated calcium/calmodulin-dependent protein kinase kinase 2(CaMKK2)-AMPK signaling pathway based on existing researches. L02 cells were induced by 100 mmol·L~(-1) ethanol for 48 h to establish the model of ALD in vitro, and 100, 50, and 25 μmol·L~(-1) concentration of MFA was treated. MiR-378b plasmids(containing the overexpression plasmid-miR-378b mimics, silence plasmid-miR-378b inhibitor, and their respective negative control-miR-378b NCs) were transfected into L02 cells by electroporation to up-regulate or down-regulate the levels of miR-378b in L02 cells. The levels of total cholesterol(TC) and triglyceride(TG) in cells were detected by commercial diagnostic kits and automatic biochemical analyzers. The expression levels of miR-378b in L02 cells were detected by real-time quantitative polymerase chain reaction(qRT-PCR). CaMKK2 mRNA levels were detected by PCR, and protein expressions of related factors involved in lipid synthesis, decomposition, and transport in lipid metabolism were detected by Western blot. The results displayed that ethanol significantly increased TG and TC levels in L02 cells, while MFA decreased TG and TC levels. Ethanol up-regulated the miR-378b level, while MFA effectively inhibited the miR-378b level. The overexpression of miR-378b led to lipid accumulation in ethanol-induced L02 cells, while the silence of miR-378b improved the lipid deposition induced by ethanol. MFA activated the CaMKK2-AMPK signaling pathway by lowering miR-378b, thus improving lipid synthesis, decomposition, and transport, which improved lipid deposition in L02 cells. This study shows that MFA improves lipid deposition in L02 cells by regulating the CaMKK2-AMPK pathway through miR-378b.
Humans
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Ethanol/toxicity*
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AMP-Activated Protein Kinases/metabolism*
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Fatty Liver
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Triglycerides
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MicroRNAs/genetics*
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Calcium-Calmodulin-Dependent Protein Kinase Kinase/genetics*
2.Determination of linarin in Yuye Jiedu granule by RP-HPLC.
Dan LU ; Gao-rong LU ; Jie-ming ZOU
China Journal of Chinese Materia Medica 2005;30(3):191-192
OBJECTIVETo determine the content of linarin in Yuye Jiedu granule.
METHODA HPLC method was developed. The chromatographic conditions are as follows Luna C18 column and acetonitrile-0.05 mol x L(-1) phosphate buffer-phosphoric acid (30:70:0.06) as mobil phase, detection wavelenth at 327 nm.
RESULTThe linear range of linarin was 0.025-0.50 microg. The average recovery was 98.7% and RSD 2.7%.
CONCLUSIONThe method is simple and accurate, with good repeatability, and can be used for determination of linarin in Yuye Jiedu granule.
Chromatography, High Pressure Liquid ; methods ; Chrysanthemum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Glycosides ; analysis ; Lonicera ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control
3.Study on p-Hydroxybenzoic Acid on Rheumatoid Arthritis Via Inhibiting NF-κB/caspase-1 Signaling Pathway
MA Yihan ; WEI Chengqiong ; XU Xiaotian ; LU Xi ; WANG Yuhui ; DUAN Xiaoqun
Chinese Journal of Modern Applied Pharmacy 2023;40(18):2519-2525
OBJECTIVE To investigate the therapeutic effect of natural phenolic compound p-hydroxybenzoic acid(HA) on adjuvant arthritis(AA) induced by the complete Freund's adjuvant(CFA), and to clarify the mechanism of HA preliminary. METHODS Apart from the normal group, all rats received 0.1 mL CFA by plantar subcutaneous injection to induce AA rats model. And rats with AA were randomized to the model group, the low-, medium-, and high-dose HA group(2.5, 5, and 10 mg·kg-1, respectively), the positive control group(indomethacine, 5 mg·kg-1). The rats in each group were orally treated with the corresponding drugs for therapeutic intervention. The volume and leg thickness of the rats in each group were recorded and an inflated articular score was obtained. Inflammation cytokine expression(TNF-α, IL-1β and IL-6) was determined by ELISA and qPCR. Radiographs and HE staining were used to observe histopathological and pathological changes in the foot. The expressions of caspase-1 and NF-κB were examined in Western blotting. RESULTS HA could significantly alleviate joint swelling in AA rat(P<0.05), inhibited the production of inflammatory factors(TNF-α, IL-1β and IL-6) from protein and mRNA levels(P<0.05), and decreased the expression levels of caspase-1 and NF-κB at protein level(P<0.05). HA alleviated ankle injury in rats by X-ray examination, and HE staining showed that HA could significantly inhibit the infiltration of inflammatory cells and the destruction of cartilage surface(P<0.05), and these results were dose-dependent. CONCLUSION HA may relieve rheumatoid arthritis by inhibiting the NF-κB/casepase-1 signaling pathway.
4. Mogroside V inhibits H202-induced oxidative damage in MIN6 cells by PI3K/Akt pathway
Meng-Jie HAN ; Juan WANG ; Guo-Xiang LIU ; Ting LI ; Lu-Wei ZHOU ; Xu CHEN
Chinese Pharmacological Bulletin 2022;38(9):1363-1368
Aim To investigate the protective effect of mogroside V on hydrogen peroxide ( H,02 )-induced oxidative stress response in mouse islet (3 cells MIN6 and the relation of its mechanism to PI3K/Akt signa¬ling pathway.Methods MIN6 cells were treated with 500 (jimol • L_1 H,(), after mogroside V,and cell via¬bility was detected by MTT.The release of reactive ox¬ygen species ( ROS) and apoptotic percentage of MIN6 cells were determined by flow cytometry.The expres¬sions of apoptosis-related factor Bel-2 , proliferation-re¬lated factor PCNA, protein Akt and p-Akt were deter¬mined by Western blot.Results H,02 restrained the proliferation of MIN6 cells obviously, induced ROS pro¬duction and apoptosis, and reduced the expression of Bel-2 and PCN A.The expressions of protein Akt and p-Akt decreased.After treatment of mogroside V , the release of ROS decreased, and the apoptosis of MIN6 cells was inhibited.The expression levels of apoptosis- related protein Bcl-2 and proliferation-related protein PCN A were reversed.The expressions of protein Akt and P-Akt increased.The viability of MIN6 cells in¬duced by H,0, increased.In addition, mogroside V partly reversed the apoptosis induction and ROS pro¬duction of Akt inhibitor MK2206 (5 jjimol • L"1 ) on MIN6 cells.Conclusions Mogroside V has protec¬tive effect on H202-induced oxidative damage in MIN6 cells and its mechanism is related to PI3K/Akt signa¬ling pathway.
5.Clinicopathological analysis of 23 cases of NK/T-cell lymphoma of nasal type.
Mingshen LU ; Fancai LI ; Qiaoyan HOU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(2):57-60
OBJECTIVE:
To improve the understanding of nasal NK/T-cell lymphoma by analyzing its phenotypic and clinicopathological features.
METHOD:
Twenty-three cases of nasal NK/T-cell lymphoma diagnosed between 2003 and 2007 in the department of pathology of Guilin Medical College were included in the study. The expression level of TIA-1, CD56, CD3, CD20, CK and EBV markers was determined by immunohistochemistry. The results were correlated with clinicopathological features.
RESULT:
69.9% (16/23) of the nasal NK/T-cell lymphoma occurred in the nasal cavity. All the 23 cases displayed necrosis, ulceration and nose bleeding. 39.1% (9/23) showed angiodestructive growth pattern. 21.74% (5/23) were accompanied by squamous cell carcinoma-like epitheliomatous hyperplasia. All the cases were positive for TIA-1 and CD3. 95.7% (22/23) of the cases were positive for CD56, while 21.7% (5/23) were weakly positive for EBV. None of the cases was positive for either CD20 or CK.
CONCLUSION
Nasal NK/T-cell lymphoma is characterized by multiple clinicopathological features. Attention is needed to differentiate the tumor from inflammatory lesions and low grade squamous cell carcinoma. Understanding of various morphological and phenotypic features (i.e. expression of TIA-1, CD56 and CD3, and lack of CD20 and CK) is the key for the diagnosis of nasal NK/T-cell lymphoma.
Adolescent
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Adult
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Aged
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Humans
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Lymphoma, Extranodal NK-T-Cell
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pathology
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Middle Aged
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Nose Neoplasms
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pathology
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Young Adult
6.Silenced ANP32A inhibits the growth, invasion and migration of colorectal cancer in vitro via the inactivation of AKT pathway.
Hong Fang DING ; Xiao Juan LI ; Lu Wei ZHOU ; Zhi CUI ; Hai De MENG ; Juan WANG
Journal of Southern Medical University 2023;43(1):52-59
OBJECTIVE:
To investigate the effect of ANP32A silencing on invasion and migration of colon cancer cells and the influence of the activity of AKT signaling pathway on this effect.
METHODS:
Colorectal cancer HCT116 and SW480 were transfected with a small interfering RNA targeting ANP32A via a lentiviral vector. At 24, 48 and 72 h after the transfection, the changes in cell proliferation and AKT activity in the cells were detected using MTT assay and Western blotting, respectively. HCT116 and SW480 cells were treated with the AKT agonist SC79 or its inhibitor MK2206 for 24, 48, 72 and 96 h, and the changes in cell migration and invasion ability were analyzed using Transwell chamber assay and cell proliferation was assessed using MTT assay. The effects of SC79 and MK2206 on migration and invasion abilities of HCT116 and SW480 cells with or without ANP32A silencing were examined using wound healing and Transwell chamber assays, and the changes in the expression of metadherin (MTDH), a factor associated with cells invasion and migration, was detected with Western blotting.
RESULTS:
Lentivirus-mediated ANP32A silencing significantly down-regulated the activity of AKT and inhibited the proliferation of both HCT116 and SW480 cells (P < 0.01). The application of AKT inhibitor MK2206 obviously inhibited the proliferation, invasion and migration of the colorectal cancer cells (P < 0.05), while the AKT agonist SC79 significantly promoted the invasion and migration of the cells (P < 0.01). In HCT116 and SW480 cells with ANP32A silencing, treatment with MK2206 strongly enhanced the inhibitory effects of ANP32A silencing on cell invasion and migration (P < 0.05) and the expression of MTDH, while SC79 partially reversed these inhibitory effects (P < 0.01).
CONCLUSION
ANP32A silencing inhibits invasion and migration of colorectal cancer cells possibly by inhibiting the activation of the AKT signaling pathway.
Humans
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Proto-Oncogene Proteins c-akt
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Cell Proliferation
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Blotting, Western
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Cell Movement
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Colonic Neoplasms
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Membrane Proteins
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RNA-Binding Proteins/genetics*
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Nuclear Proteins
7.Agreement of corneal high-order aberrations measured by three wavefront aberrometers in myopic adults
Hua-Xian ZOU ; Qi CHEN ; Li-Li LI ; Die-Feng WEI ; He-Juan MAO ; Yan-Yan HUANG ; Peng-Fei LU ; Hai-Bin ZHONG
International Eye Science 2023;23(10):1723-1731
AIM:To evaluate the agreement of corneal high-order aberrations from Topcon KR-1W, i.Profiler and OPD-Scan Ⅲ wavefront aberrometers in myopic adults.METHODS:A prospective clinical study. A total of 92 adult patients(92 eyes)with myopia in the department of optometry, the People's Hospital of Guangxi Zhuang Autonomous Region from June to August 2022 were enrolled. The third-order and fourth-order corneal aberrations at the pupil diameter of 4 and 6mm were measured by Topcon KR-1W, i.Profiler, and OPD-Scan Ⅲ, respectively. The difference and agreement of the three aberrometers were evaluated.RESULTS: The measurements at 6mm pupil diameter were all greater than those at 4mm pupil diameter. Although there were no statistical differences in the measurements of Z-44、Z-24 by the three aberrometers at 4 pupil diameter(P>0.05), there were statistical differences in other measurements(P<0.05). The aberration results measured by the three aberrometers were statistically different at the 6mm pupil diameter(P<0.05). The 95% limit of agreement(95%LoA)of the measurements of higher-order aberration, including the third-order aberrations at 4mm pupil diameter and the third-order and fourth-order aberrations at 6mm pupil diameter(except for the Z-24)were greater than 0.1μm. The concordance correlation coefficient(Pc)was lower than 0.90, indicating a poor consistency. The correlation coefficients of corneal higher-order aberrations were significantly different among the three aberrometers at 4 and 6mm pupil diameter(r4mm=0.215~0.805, P4mm<0.05; r6mm=0.561~0.916, P6mm<0.001).CONCLUSION:There were significant differences in the measurements of the third- and fourth-order corneal aberrations at 4 and 6mm pupil diameter among Topcon KR-1W, i.Profiler, and OPD-Scan Ⅲ, and the agreements were poor, so they are not interchangeably in clinical applications.
8.Effect of early inhibition of TLR4 on hippocampal immune function to adolescence after neonatal HIBD
Xiaoli Huang ; Zhicui Ouyang ; Xianghong Wu ; Yan Li ; Yun Huang ; Guoqiong Liu ; Shiwei Lu ; Zhen Tang
Acta Universitatis Medicinalis Anhui 2023;58(8):1317-1322
Objective :
To investigate the role of early inhibition of Toll⁃like receptor 4 (TLR4) in regulating hippampal neuroimmune responseto adolescent ratsafter neonatal hypoxic⁃ischemic brain damage(HIBD) .
Methods:
Postnatal day 7 rats were randomized into controlgroup , hypoxic ischemia (HI) group , and HI + TAK⁃242( the specific inhibitor of TLR4)(TAK⁃242) group. The expression of TLR4 in rat hippocampus was detected by immunohistochemistry at 3 days after HI. Immunofluorescence were used to determine the number of Iba⁃1 + , GFAP + , CD161 + , MPO + and CD3 + cells in the hippocampus at 21 days after HI. Immunohistochemistry was used to detect ICAM⁃1 and C3a expression in the hippocampal CA1 region ; and Western blot was used to detect tumor necrosis factor interleukin IL⁃1β , TNF⁃α and IL⁃10 expression.
Results :
Compared with control group , significantly raised TLR4 expression was observed in the left hippocampal CA1 , CA3 and DG regions(P < 0. 01 or P < 0. 05) , while the expression in the TAK⁃242 group lowered compared to the HI group (P < 0. 05) . The number of GFAP + cells in the CA1 area of the hippocampus in the TAK⁃242 group of neonatal rats decreased compared to which in the HI group at 21 days after HI(P < 0. 05) , but the number of CD3 + T lymphocytes in the hippocampal CA1 area of new born rats in the HI group increased compared to which in the Control group (P < 0. 05) , but the difference between TAK⁃242 and the Control group was not statistically significant. The number of Iba⁃1 + cells , MPO + cells , CD161 + cells , the expression of ICAM⁃1 and C3a in hippocampal CA1 region , and the expression of TNF⁃α , IL⁃1β and IL⁃10 in hippocampus of rats were not different among groups at 21 days after HIBD.
Conclusion
Early inhibition of TLR4 may ameliorate adolescent neuroimmune disorders by reducing the increase of hippocampal astrocytesafter neonatal HIBD.
9.Study on effect of Cordyceps sinensis and artemisinin in preventing recurrence of lupus nephritis.
Chinese Journal of Integrated Traditional and Western Medicine 2002;22(3):169-171
OBJECTIVETo observe the effect of Cordyceps sinensis and artemisinin in preventing recurrence of lupus nephritis (LN).
METHODSSixty-one LN patients, who had no activities by corticosterone and cyclophosphamide (CTX) impacting therapy were randomly divided into two groups. The 31 cases in the treated group were given Cordyceps powder 2-4 g/d before meal and artemisinin 0.6 g/d after meal in three portions orally taken for 3 years. The 30 patients in the control group were treated with tripterygiitotorum and/or Baoshenkang tablet. The consecutive observation lasted for 5 years to monitor the clinical manifestations of lupus and laboratory indexes including blood creatinine, creatinine clearance rate (CCr) and antinuclear antibodies (ANA).
RESULTSThe therapeutic effect showed markedly effective in 26 cases (83.9%), effective in 4 (12.9%) and ineffective in 1 (3.2%) in the treated group, while in the control group, the corresponding numbers were 15 (50.0%), 8 (26.7%) and 7 (23.3%), the difference between the two groups in markedly effective rate was significant (P < 0.01). In the treated group, C3 level was stabilized at above 1.21 +/- 0.20 g/L, which was over the normal range, CCr was unchanged as compared before and after treatment, which was significantly different from that in the control group. Moreover, the side-effects occurred in the treated group was less.
CONCLUSIONCordyceps and artemisinin could prevent the recurrence of LN and protect kidney function.
Adult ; Aged ; Anti-Infective Agents ; therapeutic use ; Artemisinins ; therapeutic use ; Complement C3 ; metabolism ; Cordyceps ; Creatinine ; blood ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lupus Nephritis ; drug therapy ; Male ; Middle Aged ; Phytotherapy ; Secondary Prevention ; Sesquiterpenes ; therapeutic use
10.Rapid molecular assay and strain typing methods of extrapulmonary tuberculosis
Jian GUO ; Qiwen FAN ; Shuihua LU ; Guilin DENG ; Wenjuan WU
Chinese Journal of Laboratory Medicine 2013;36(7):620-624
[Abstract] Objective To evaluate the effect of Mycobacterium tuberculosis Direct Assay (MTD) for rapid detecting Mycobacterium tuberculosis rRNA and Multi-locus PCR for M.bovis BCG strain typing in patients with suspected extra-pulmonary tuberculosis.Methods From June 2010 to December 2011,47 children and 75 adult patients with suspected extra-pulmonary tuberculosis in Shanghai public health clinical center were recruited.Also 48 non-tuberculosis patients were taken as a negative control.Clinical specimens from these patients were collected.Acid fast stain,solid culture,liquid culture,and MTD were used to detect all clinical specimens simultaneously.Screen tuberculosis strains of the culture isolates by MPT64 antigen assay and use Multi-locus PCR for the BCG strain genotyping of the isolates without MPT64 antigen.SPSS16.0 was used to analyse the results.Results The sensitivity for acid fast stain,solid culture,liquid culture and MTD test was 10.7% (13/122),11.5% (14/122),16.4% (20/122) and 37.7% (46/122),respectively.And the specificity of MTD was 100.0%.Six clinical isolates from children were identified as BCG by Multi-locus PCR typing,the same with chemical tests.Conclusions The MTD assay and the MGIT960 liquid culture are effective and reliable method for diagnosing extra-pulmonary tuberculosis.And Multi-locus PCR can be assisted for the early diagnosis of extra-pulmonary tuberculosis patients with suspected BCG infection.