1.Determination of Eucalyptol in Chimonanthus Salicifolius S. Y. Hu by GC
Weiying WANG ; Juhua MAO ; Huali YU ; Le YU ; Kejun CHENG
China Pharmacist 2015;(1):154-155
Objective:To establish the quantitative method for eucalyptol in Chimonanthus salicifolius S. Y. Hu by GC. Methods:The GC method was performed on a Zebron ZB-WAX column (60 m × 0. 32 mm,0. 5 μm) with programmed temperature of 60-200℃, an FID detector was used, the detector temperature was 250℃, the inlet temperature was 220℃, and the carrier gas was nitrogen with high purity. Results:A good linearity of eucalyptol was within the range of 0. 012 6-0. 503 4 mg·ml-1(r=0. 999 8), and the average recov-ery was 100. 68%(RSD=1. 51%,n=6). Conclusion:The method is simple, accurate and quick with good reproducibility, and suitable for the quality control of Chimonanthus salicifolius S. Y. Hu.
2.Study on Methodolgy of Content Determination of Three Flavonoids in Traditional She Medicine Shi Liang Cha
Juhua MAO ; Weiying WANG ; Le YU ; Kejun CHENG
China Pharmacist 2014;(11):1808-1810
Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Shi Liang Cha. Methods:HPLC was applied with the chromatographic conditions as follows: the chromatographic column was Agilent Zorbax SB-C18 (250 mm × 4. 6 mm, 5μm) at 30℃;acetonitrile-0. 1% H3 PO4 solution was used as the mobile phase with gradient elution; the flow rate was 1. 0 ml·min-1, and the detection wavelength was 360nm. Results: Good linearity of rutin, quercetin and kaempferol was within the range of 0.0409-1.637 0mg·ml-1(r=0.999 2), 0.44-88.00μg·ml-1(r=0.999 8) and 0.41-77.63μg·ml-1(r=0. 999 2), respectively; the average recovery was 99.35%(RSD =1. 64%),101. 14% (RSD =1. 88%) and 99. 69% (RSD =1. 92%) , respectively. Conclusion:The method is simple, accurate and repeatable, and suitable for the simultaneous determination of rutin, quercetin and kaempferol in Shi Liang Cha.
3.Study on chemical constituents of Drosera peltata var. multisepala.
Lin LI ; Jin HUANG ; Xianghua XU ; Yao ZHANG ; Kejun CHENG ; Peizhong YU
China Journal of Chinese Materia Medica 2012;37(2):222-225
Chemical investigatation of Drosera peltata var. multisepala led to the isolation of eleven compounds using various chromatographic techniques. The structures of these compounds were elucidated as isoshinanolone-4-O-beta-D-glucoside (1), isoshinanolone (2), epi-isoshinanolone (3), plumbagin (4), droserone (5), droserone-5-O-glucoside (6), quercetin (7), kaempferol (8) , gossypetin-8-O-glucoside (9), 3,3'-dimethoxy ellagic acid (10), and ellagic acid (11) by their physicochemical properties and spectral data analysis. Compound 1 was a new compound. Compounds 3, 8, 10, and 11 were isolated from this plant for the first time.
Chromatography, Liquid
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methods
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Drosera
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chemistry
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Drugs, Chinese Herbal
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chemistry
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Ellagic Acid
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analogs & derivatives
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analysis
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isolation & purification
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Glucosides
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analysis
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isolation & purification
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Kaempferols
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analysis
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isolation & purification
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Magnetic Resonance Spectroscopy
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methods
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Molecular Structure
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Naphthoquinones
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analysis
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isolation & purification
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Plant Extracts
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analysis
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isolation & purification
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Quercetin
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analysis
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isolation & purification
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Spectrometry, Mass, Electrospray Ionization
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methods
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Tetrahydronaphthalenes
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analysis
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isolation & purification
4.mRNA Expression of Core Molecules in the Downstream of mTOR Pathway and Its Relationship to Clinical ;Features in Primary Acute Myeloid Leukemia
Kejun CHEN ; Hui CHENG ; Jianmin NG WA ; Jianmin NG YA ; Shuqing LÜ
Chinese Journal of Clinical Medicine 2016;23(1):17-21
Objective:To explore the relationship between the mRNA expression of mammalian target of rapamycin (mTOR) , eukaryotic translation initiation factor 4E (eIF‐4E) and eukaryotic translation initiation factor 4E binding protein 1 (4E‐BP1) in bone marrow mononuclear cells of primary acute myeloid leukemia (AML) patients and clinical features of AML .Methods :The mRNA expression levels of 4E‐BP1 ,eIF‐4E and mTOR in bone marrow mononuclear cells were detected in 59 patients with primary AML and 15 normal human as a control group by reverse transcription‐polymerase chain reaction (RT‐PCR) . Results:The percentages of mTOR ,eIF‐4E and 4E‐BP1 mRNA high expression in 59 cases of AML were 50 .9% ,59 .4% and 45 .8% ,respectively .The platelet count in high 4E‐BP1 mRNA expression group was significantly lower than that in low group (P=0 .044) .The percentage of bone marrow blast cells in high eIF‐4E mRNA expression group was higher than that in low group ,but there was no significant difference between the two groups (P= 0 .068) .The percentage ,and extramedullary invasion of bone marrow blast cells in high mTOR expression group was significantly higher than that in low group (P=0 .040 , P=0 .029) .The percentage of anemia in high mTOR expression group was higher than that in low group ,but there was no significant difference between the two groups (P=0 .082) .Conclusions :Core protein mRNA in mTOR pathway downstream were closely relative to the proliferation of bone marrow blast cells ,PLT count ,anemia and extramedullary invasion in AML . They can help to choose effective and safe chemotherapy regimen ,as well as to prevent side effects of chemotherapy such as anemia and bleeding .
5.Effect of hybrid intensity modulated radiotherapy on the immune function of patients with locally advanced breast cancer surgery and efficacy observation
Kejun DAI ; Xujing LU ; Xifa ZHOU ; Mingming FANG ; Ling CHEN ; Jun LIU ; Yuqiong DING ; Cheng GU
Cancer Research and Clinic 2020;32(11):766-771
Objective:To investigate the effect of hybrid intensity modulated radiotherapy (Hy-IMRT) on immune function in patients with locally advanced breast cancer surgery and the treatment efficacy.Methods:A total of 94 patients with locally advanced breast cancer who underwent modified radical mastectomy for breast cancer and required postoperative radiotherapy in Changzhou Cancer Hospital in Jiangsu Province from January 2015 to January 2017 were selected. The patients were divided into Hy-IMRT group (observation group, 47 cases) and three-dimensional conformal radiotherapy (3DCRT) group (control group, 47 cases) according to the random number table method. The dose and related radiophysical parameters of the respective target areas of the two groups, adverse reactions during and after radiotherapy, cytokines and T lymphocyte subsets before and after radiotherapy, 3-year local recurrence rate, distant metastasis rate and mortality were observed and compared between the two groups.Results:The dose obtained by 95% (D 95%) [(4 945.6±36.1) Gy vs. (4 754.0±35.6) Gy] and target area conformity (CI) of the target volume (0.7±0.1 vs. 0.5±0.1) in the observation group were greater than those in the control group, and the differences were statistically significant (both P<0.05); the target volume of 110% of the prescription dose (V 110%) [(1.6±0.5) cm 3 vs. (8.4±1.2) cm 3], the target volume of more than 105% of the prescription dose (V 105%) [(19.3±3.5) cm 3 vs. (26.6±5.6) cm 3] and the heterogeneity index (HI) (1.1±0.1 vs. 1.3±0.1) in the observation group were all smaller than those of the control group, and the differences were statistically significant (all P < 0.05). The incidence of acute skin adverse reactions [53.2% (25/47) vs. 74.5% (35/47)] and the incidence of bone marrow suppression [40.4% (19/47) vs. 70.2% (33/47)] in the observation group were lower than those in the control group, and the differences were statistically significant (both P < 0.05). There were no significant differences in levels of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), CD4 +, CD8 +, and CD4 +/CD8 + between the two groups before radiotherapy (all P > 0.05). At the end of radiotherapy, the levels of IL-6, TNF-α and CD8 + were higher in both groups than before radiotherapy (all P < 0.05), and CD4 + and CD4 +/CD8 + were lower than before radiotherapy (both P < 0.05). The levels of IL-6, TNF-α and CD8 + in the observation group were lower than those in the control group, while the CD4 + and CD4 +/CD8 + were higher than those in the control group (all P < 0.05). The 3-year local recurrence rate [34.04% (16/47) vs. 42.55% (20/47)], distant metastasis rate [25.53% (12/47) vs. 38.30% (18/47)] and mortality rate [14.89% (7/47) vs. 19.15% (9/47)] in the observation group were lower than those in the control group, but the differences were not statistically significant (all P > 0.05). Conclusion:Compared with 3DCRT, the Hy-IMRT has less effect on the immune function of locally advanced breast cancer patients after modified radical resection, and the incidences of acute skin reaction and bone marrow adverse reaction are low.
6.Qingshen granules regulate mitophagy through miR-4516/SIAH3/PINK1 to attenuate renal fibrosis in rats
Kejun REN ; Hua JIN ; Dong WANG ; Yanping MAO ; Rong DAI ; Meng CHENG ; Yiping WANG ; Lei ZHANG
Chinese Journal of Pathophysiology 2024;40(8):1479-1487
AIM:To explore the mechanism by which Qingshen granules(QSG)intervene in the microRNA-4516(miR-4516)targeted regulation of the SIAH3/PINK1 axis,enhancing mitophagy and inhibiting renal fibrosis.METHODS:Male SD rats were randomly divided into three groups:control,model,and QSG groups.The QSG aqueous solution was administered via gavage once daily,4 mL each time,for 8 consecutive weeks.Blood creatinine levels were measured in each group.Hematoxylin-eosin(HE)and Masson staining were utilized to assess the degree of renal patholog-ical damage.Western blot analysis was performed to determine the expression levels of β-actin,PINK1,Parkin,SIAH3,VDAC1,Mfn1,Mfn2,OPA1,LC3B,and P62 proteins in renal tissue.RT-qPCR was used to detect the mRNA expres-sion level of SIAH3 in rat kidney tissue,and transmission electron microscopy was employed to observe mitochondrial dam-age in renal tissue.QSG-containing serum and transforming growth factor-β1(TGF-β1)were used to induce an HK-2 cell fibrosis model.The cells were divided into the following groups:normal cell(NC)group,model cell(MC)group,MC+miR-4516 mimics group,MC+miR-4516 NC+QSG group,MC+miR-4516 mimics+QSG group,and MC+QSG group.Cell activity in each group was detected using the CCK-8 method,and Western blot analysis was performed to determine E-cad-herin and α-SMA protein expression levels.The regulation of SIAH3 by miR-4516 was verified using a dual luciferase re-porter assay.RT-qPCR was used to detect the mRNA expression of miR-4516,SIAH3 mRNA,and PINK1.RESULTS:The results indicated that QSG intervention reduced fibrosis in rat renal tissue and HK-2 cells,decreased SIAH3 mRNA expression,increased PINK1 expression,and activated mitophagy in renal tissue.In vitro results confirmed that QSG can elevate miR-4516 expression,inhibit SIAH3 mRNA expression,promote PINK1 expression in HK-2 cells,and reduce the expression of the fibrosis marker protein α-SMA.CONCLUSION:In summary,this study preliminarily clarified the mechanism by which QSG intervention targets miR-4516 to regulate the SIAH3/PINK1 axis,thereby enhancing mitophagy and inhibiting renal fibrosis.
7.Study on the Content Determination and Accumulation Law of Saponins in Paris Polyphylla Smith Var. Chinensis (Franch.) Hara
Jie FANG ; Wenliang CHENG ; Junmei ZHOU ; Junjie PAN ; Wei CHENG ; Qundan LYU ; Kejun CHENG
Chinese Journal of Modern Applied Pharmacy 2024;41(2):236-241
OBJECTIVE
To analyze the accumulation law of saponins during growth in Paris polyphylla Smith var. chinensis (Franch.) Hara, determine the content of the main saponins in different cultivation years, age groups, cultivation modes and provenances.
METHODS
The content of 5 kinds saponins(I, II, VI, VII, H) was simultaneous determined by HPLC.
RESULTS
The total saponins in P. polyphylla Smith var. chinensis (Franch.) Hara were mainly composed of saponin VII and H, supplemented by saponin VI, I and II. The content of saponins(I, II, VII) was significantly different among different cultivation years rhizome, while it reached the standard of Chinese Pharmacopoeia 2020 Edition after 6 years old, 8-year-old rhizome was the highest. The saponins(I, II, VII) content in 4a rhizome and 5a rhizome was significant higher than others, and it ranged from 0.354% to 0.765% in different cultivation modes, from high to low as follows: coniferous forest>bamboo forest>broadleaf forest>greenhouse. In different provenances, it ranged from 0.592% to 0.741%, reached the highest level in Qingyuan Baikan, and was slightly lower than the standard of Chinese Pharmacopoeia 2020 Edition in Sanming Fujian.
CONCLUSION
There are remarkable correlations among saponins accumulation amounts and cultivation years, age groups, cultivation modes and provenances, which can provide reference for the artificial culvication of P. polyphylla Smith var. chinensis (Franch.) Hara.
8.Determination and Comparison of 26 Components in Different Parts of Two Base Plants of Shiliang Tea Based on UHPLC-MS/MS
Juhua MAO ; Junjie PAN ; Zhangjin CHEN ; Chaqing WU ; Weiying WANG ; Kejun CHENG
Chinese Journal of Modern Applied Pharmacy 2024;41(4):489-495
OBJECTIVE
To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS) method for determination and comparison of 26 components in different parts of two base plants of Shiliang tea(Chimonanthus salicifolius S.Y.Hu and Chimonanthus zhejiangensis M.C.Liu), and screen quality markers of different parts.
METHODS
The UHPLC method was performed on an Agilent RRHD Eclipse Plus C18 (2.1 mm×50 mm, 1.8 μm) column with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.3 mL·min−1, the column temperature was 35 ℃, and the injection volume was 0.5 μL; the multiple reaction monitoring mode was employed for the quantification of 26 components with electrospray ionization(ESI) source polarity in negative and positive mode.
RESULTS
Good linear relationship(r >0.999) were observed in the test ranges for 26 compounds, and the average recovery was 88.5%−111.7% with RSD was 3.4%−9.8%. There was no significant difference between the two base plants of Shiliang tea, and all of these samples were divided into two categories by hierarchical cluster analysis. The main components in leaves was flavonoids, among them, the content of kaempferol 3-O-rutinoside was the highest, reaching 12.902 mg.g−1; the main components in stems and roots was coumarins, and the content of alkaloids in roots was higher, relatively; 7 quality markers of difference were screened by OPLS-DA, which were kaempferol 3-O-rutinoside, chimonanthine, rutin, fraxetin, calycanthoside, scopolin, neochlorogenic acid.
CONCLUSION
These study elucidates the differences of chemical components in the different parts of two base plants of Shiliang tea, which providing basis for the research of pharmacodynamic substances and references for the comprehensive utilization of Chimonanthus salicifolius S.Y. Hu and Chimonanthus zhejiangensis M.C.Liu resources.
9.Expression of heme oxygenase-1 in nasal polyps and regulation by glucocorticoid.
Yu WANG ; Zhijian YU ; Jun SHI ; Lan CHENG ; Kejun ZUO ; Guozhen MENG ; Weiping WEN ; Huabin LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2016;51(3):169-173
OBJECTIVETo evaluate the expression and possible modulation of heme oxygenase-1 (HO-1) in nasal polyps of patients with chronic rhinosinusitis with nasal polyps (CRSwNP).
METHODSNasal polyps and uncinate process tissues were collected from 25 CRSwNP patients and 19 healthy controls with nasal septal deviation. HO-1 expression was examined using qRT-PCR, immunohistochemistric staining and Western blot analysis. Moreover, additional uncinate process mucosal samples of 15 healthy controls with nasal septal deviation were harvested for nasal explant culture experiments. HO-1 expression was measured in cultured nasal explant in response to specific inflammatory and glucocorticoid stimulation. SPSS 20.0 software was used to analyze the data.
RESULTSThe mRNA and protein expression of HO-1 was significantly increased in polyp tissues, 1.220±0.397 in mRNA and 1.409±0.701 in protein, compared with healthy controls 0.464±0.318 in mRNA and 0.017±0.1147 in protein (U=22.00 in mRNA and U=1.00 in protein, both P< 0.05). The immunohistochemical results showed that HO-1 was mainly distributed in the epithelial layer, submucosal glands and inflammatory cells in nasal tissues. Nasal explant culture experiments demonstrated that HO-1 mRNA was upregulated by IL-17A. The HO-1 mRNA level before the stimulation was 1.000, and 17.264±4.275 after the stimulation of 1 ng/ml IL-17A (U=0, P<0.05), 19.128±4.605 after the stimulation of 10 ng/ml IL-17A (U=0, P<0.05), but was significantly suppressed after stimulation with glucocorticoids (dexamethasone, DEX). The mRNA level after the glucocorticoids stimulation was 0.370±0.101 (U=0, P<0.05) and 0.316±0.167 (U=0, P<0.05) respectively. Furthermore, the HO-1 mRNA was inhibited by TGF-β1, the mRNA level was 0.217±0.322 (U=0, P<0.05), 0.070±0.070 (U=0, P<0.05), respectively.
CONCLUSIONIncreased HO-1 expression may play a role in the pathogenesis of CRSwNP, which may be considered as the therapeutic target.
Blotting, Western ; Case-Control Studies ; Chronic Disease ; Dexamethasone ; pharmacology ; Glucocorticoids ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Humans ; Interleukin-17 ; pharmacology ; Nasal Polyps ; complications ; metabolism ; RNA, Messenger ; metabolism ; Rhinitis ; complications ; metabolism ; Sinusitis ; complications ; metabolism ; Tissue Culture Techniques ; Transforming Growth Factor beta1 ; metabolism
10.Research progress on the pathological mechanism of meibomian gland dysfunction in diabetic patients
Yize HAN ; Kejun LI ; Qingmin MA ; Fang FAN ; Wenwen TANG ; Jing CHENG ; Linwei YAN ; Yafeng WANG
International Eye Science 2024;24(7):1098-1101
Meibomian gland dysfunction is a chronic and diffuse disease of the meibomian glands, characterized by obstruction and(or)abnormal secretion of the terminal ducts. Clinically, it can lead to tear film abnormalities and inflammation of the ocular surface, resulting in symptoms of ocular irritation and potential corneal damage that may impact visual function. Meibomian gland dysfunction can be classified into two types based on meibomian gland secretion: low secretion type and high secretion type. The low secretion type further includes acinar atrophy type and obstruction type. In recent years, research has revealed that patients with diabetes experience chronic damage to their meibomian gland tissue in the early stages of the disease, leading to structural and functional changes. The incidence and severity of meibomian gland dysfunction are higher in diabetic patients. However, there are numerous complex factors contributing to this condition in diabetes patients, and mechanisms remain unclear at present. This article reviews both domestic and international research progress on the pathological mechanism underlying meibomian gland dysfunction in diabetes.