1.Gas Chromatographic Fingerprint of Oleic Acid of Whitmania pigra Whitman
Yong HUANG ; Zhili ZUO ; Yilin YANG ; Yueyun ZHANG ; Fan WEI ; Chengjian ZHAO ; Jianhua MIAO
Journal of Guangzhou University of Traditional Chinese Medicine 2016;33(5):716-719
Objective To establish the gas chromatographic(GC) fingerprint of oleic acid of Whitmania pigra Whitman for its quality control. Methods Ten batches of Whitmania pigra from different sources and processed by different methods were analyzed with Agilent 6890N gas chromatography detector on DB-WAX(30 mm × 0.32 mm × 0.25μm)column at the vaporizing temperature of 270℃, column temperature of 130℃and flame ionization detector (FID) temperature of 280℃. We used a software of Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version of 2004A) published by Chinese Pharmacopoeia Commission to calculate GC similarity. Results Oleic acid content of Whitmania pigra processed by different methods had significant differences (F2,7 = 7.350, P = 0.019). The oleic acid content of samples dried after washing with clean water significantly differed from that of the samples processed by alumen or the slices dried naturally(P = 0.021, P= 0.009). The similarity of the fingerprints was in the range of 0.458 - 0.998. The similarity of samples from Lipu of Guangxi Province was the lowest. Conclusion The fingerprints of most of the samples have very high similarity. The established GC fingerprints can be used to effectively identify the qualified or inferior Whitmania pigra products, which will provide some reference for the quality control of Whitmania pigra.
2.Role of homocysteine to promote the vascular smooth muscle cell proliferation by MiR-125 b methylation
Xianmei LIU ; Chengjian CAO ; Jue TIAN ; Li ZHAO ; Fanqi KONG ; Longxia ZHOU ; Jiukai CHEN ; Yanhua WANG ; Xiaoling YANG ; Yuexia JIA ; Yideng JIANG
Chinese Pharmacological Bulletin 2015;(7):1023-1027
Aim To investigate the role of miR-125 b and its DNA methylation in homocysteine ( Hcy )-in-duced vascular smooth muscle cells( VSMCs) prolifera-tion. Methods VSMCs were stimulated with 0,50, 100, 200, 500 μmol · L-1 Hcy respectively. Then qRT-PCR was used to detect the mRNA levels of miR-125b,and nested-touchdown methylation-specific PCR ( ntMS-PCR) was used to detect the methylation levels of miR-125b. VSMCs were transfected with miR-125b precursor or the inhibitor of miR-125b ,then 3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide ( MTT ) assay was used to reflect the proliferation of VSMCs. The distribution of CpG islands of miR-125b promoter region was analyzed by bioinformatics meth-ods. VSMCs were stimulated with 100 μmol·L-1 Hcy and transfected with or without DNA methylation inhib-itors 5-nitrogen impurity cytidine ( AZC) , then the ex-pression of miR-125b was detected by qRT-PCR. Re-sults The mRNA levels of miR-125 b were decreased in 100,200,500 μmol·L-1 Hcy group compared with 0 μmol·L-1 Hcy group. The precursor of miR-125b could inhibit the proliferation activity and the inhibitor of miR-125 b could increase the proliferation activity of VSMCs cells. Bioinformatics analysis indicated that MiR-125 b promoter region had a CpG island whose length was 792 bp ( 1881-2672 ) . The miR-125 b pro-moter region methylation levels increased after Hcy in-tervention ( P <0. 01 ) . The expression level of miR-125 b increased after AZC intervention ( P <0. 05 ) . Conclusions ① Hcy promotes vascular smooth mus-cle cell proliferation maybe by down-regulating the ex-pression of miR-125b. ② Hcy down-regulates the ex-pression of miR-125 maybe by up-regulating the methy-lation levels of miR-125b promoter region.
3.Role of ERO1αand its DNA methylation in homocysteine-induced inhibition of hepatocyte proliferation
Li ZHAO ; Chengjian CAO ; Xianmei LIU ; Fanqi KONG ; Wenbin MA ; Longxia ZHOU ; Jiukai CHEN ; Minghao ZHANG ; Yun JIAO ; Xiaoling YANG ; Yideng JIANG
Chinese Pharmacological Bulletin 2014;(12):1743-1747
Aim To explore the role of ERO1 αand its DNA methylation in homocysteine (Hcy)-induced in-hibition of hepatocytes proliferation.Methods The hepatocytes stimulated with 0 μmol·L -1 Hcy were set as the normal group (NC group)and the hepatocytes stimulated with 1 00 μmol·L -1 Hcy as the experimen-tal group (Hcy group).Methyl thiazolyl tetrazolium (MTT)reduction assay was used to reflect the prolifer-ation of the hepatocytes;qRT-PCR and Western blot were used to detect the mRNA and protein levels of ERO1 α;the expression of green fluorescence protein was observed in hepatocytes after the recombinant plas-mid of ERO1 α was constructed,which was used to confirm if the recombinant plasmid into hepatocytes was successful,then the mRNA and protein levels of ERO1 αwere assayed and the proliferation of the hepa-tocytes was also detected;ntMSP was used to detect the change of ERO1 αDNA methylation.Results The mRNA and protein levels of ERO1 αwere decreased in Hcy group compared with NC group,and the prolifera-tion activity of hepatocytes in Hcy group was de-creased.Sequencing result showed that the recombi-nant plasmid of ERO1 αwas constructed successfully. QRT-PCR and Western blot revealed that ERO1 αwas overexpressed. The result of MTT suggested that ERO1 αoverexpression restored hepatocyte proliferation inhibited by Hcy.Hcy caused ERO1 αDNA hyperm-ethylation.Conclusions Hcy inhibits hepatocyte pro-liferation by downregulating the expression of ERO1 α, and methylation of ERO1 αpromoter may play a role in this process.
4.Content Determination of Cantharidin in Lytta caraganae by HPLC
Tengjiao WANG ; Chengjian ZHAO ; Yingle GU ; Yongli XU ; Li LI
China Pharmacy 2018;29(7):930-932
OBJECTIVE:To establish the method for content determination of cantharidin in Lytta caraganae,and to use the result as the extract screening evidence of L. caraganae source material. METHODS:Ultrasonic extraction method was used to extract cantharidin from L. caraganae using acetone as solvent. HPLC method was adopted to determine the content of cantharidin. The determination was performed on C18column with mobile phase consisted of methanol-water(23:77)at flow rate of 1.0 mL/min. The detection wavelength was set 230 nm,the column temperature was set at 35 ℃,and sample size was 10 μL. The content of cantharidin in L. caraganae was determined and compared with the results of content determination by the method stated in Chinese Pharmacopeia(using chloroform as extraction solvent). RESULTS:The liner range of cantharidin were 0.2-1.0 mg/mL (r=0.998 8)with average methodology recovery rates of 101.1%(RSD=1.7%,n=6). The average content of cantharidin in L. caraganae was 0.932%(n=3),while the content of cantharidin was 0.793%(n=3)determined by the method stated in Chinese Pharmacopeia. Both were higher than the requirement of Chinese Pharmacopeia that the content of cantharidin in scource material of cantharidin was higher than 0.35%. CONCLUSIONS:Established method is accurate and reliable for the content determination cantharidin in L. caraganae. The content of cantharidin is up to the standard of Chinese Pharmacopeia,and can be used as source material for exacting cantharidin.
5.Clinical application of preset balloon in abdominal aorta in assisting cesarean section for patients with placenta previa complicated by placenta accreta
Hongyu WANG ; Chengjian SUN ; Yanhua WANG ; Liming SHAO ; Shui YU ; Xiaolong ZHAO ; Haofei TAN
Journal of Interventional Radiology 2017;26(10):922-926
Objective To investigate the clinical value of preset balloon in abdominal aorta in assisting cesarean section for patients with placenta previa complicated by placenta accreta.Methods Preset balloon in abdominal aorta was performed in 72 patients with placenta previa complicated by placenta accreta before cesarean section was carried out.Of the 72 patients,pernicious placenta previa (type A) was seen in 33,scar uterus,placenta previa with placenta accreta (type B) in 36 patients,and placenta previa complicated by placenta accreta with no cesarean section history (type C) in 3.The amount of blood loss during cesarean section,the course of uterine resection,the X-ray radiation dose in performing placement of balloon,and the procedure-related complications were recorded.Results The average amount of intraoperative blood loss in type A,B and C group was 1461 ml,947 ml and 533 ml,respectively.Subtotal hysterectomy was adopted in 9 patients and uterine repair in 32 patients.The average radiation doses in 17 patients who received preset balloon procedure in 2015 and in 55 patients who received preset balloon procedure in 2016 were (28.5±14.1) mGy and (3.7±2.5) mGy,respectively.During hospitalization period,one patient developed right superficial femoral artery thrombosis,one patient developed venous thrombosis of right lower extremity,and subcutaneous hematoma occurred in two patients.Conclusion Temporary occlusion of the abdominal aorta with preset balloon to assist the cesarean section for patients with placenta previa complicated by placenta accreta can effectively reduce the amount of intraoperative blood loss and markedly reduce hysterectomy rate.The procedure of preset balloon is simple,and the X-ray exposure time is short.Skilled and experienced manipulation can further reduce the incidence of complications.
6.Quantitative analysis of five compounds in gallnut lotion by UPLC-MS/MS
Qun ZHANG ; Yu CHEN ; Xiaowei ZHI ; Qiuping XU ; Yingnan YANG ; Liang ZHAO ; Chengjian LI
Journal of Pharmaceutical Practice 2021;39(5):437-441
Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously detect the contents of gallic acid, syringin, phellodendrine, aesculin and rhein in the gallnut lotion. Methods An UPLC- MS/MS method was established. Separation was performed on an Agilent Poroshell 120 EC-C18(2.1 mm×150 mm, 2.7 μm)with a gradient mobile phase system of 0.2% formic water-acetonitrile solution. The flow rate was 0.3 ml/min. The temperature of column was 30 ℃. The injection volume was 2 μl. The MS detection was in dynamic MRM mode. Results gallic acid, syringin, phellodendrine, aesculin and rhein were successfully separated using this method, with good linear relationship as the ranges of 153.8-15380、10.31-1031、5.265-526.5、50.70-5070、1.054-105.4 ng/ml, respectively. The precision, repeatability, stability and recovery were good. Conclusion This UPLC-MS/MS method is stable, rapid, and reproducible., It is suitable for detecting the contents of gallic acid, syringin, phellodendrine, esculetin and in the gallnut lotion.
7.Radiation level analysis of some non-medical institutions in Hangzhou
Zhixin ZHAO ; Qiang WANG ; Chengjian CAO ; Yong YANG ; Haihua WANG ; Hezheng ZHAI
Chinese Journal of Radiological Health 2021;30(1):59-62
Objective The purpose of this study is to grasp the current situation of radiation protection in some non-medical institutions in Hangzhou, and to provide basis and reference for the relevant authorities departments to make the radioactive hazard factors monitoring plan for non-medical institutions. Methods The configuration of the personnel protective equipment and radiation level of radiation sources and radiation devices in 5 non-medical monitoring institutions in Hangzhou were investigated and analyzed by means of questionnaire survey and on-site detection. Results The radiation workers of 5 monitoring institutions have carried out personal dose monitoring, and their annual individual dose equivalent meets the requirements of relevant national standards. The ambient dose equivalent rates around the radiation source and radiation device room are lower than the minimum detection limit of the instrument. When the source is stored, the maximum ambient dose equivalent rates at 5 cm and 100 cm away from the external surface of the source container were 22.2 μSv/h and 2.0 μSv/h, respectively. When the source is in use, the maximum ambient dose equivalent rates at 5 cm and 100 cm away from the external surface of the source container were 44.3 μSv/h and 5.0 μSv/h, respectively. Conclusion Under normal operation, the radiation dose level of some non-medical monitoring institutions in Hangzhou is at a low level, while the ambient dose equivalent rates around the external surface of the source container is at a high level. Therefore, non-medical institutions should improve their awareness of radiation protection and strengthen radiation management.
8.Inhibition of ebracteolatain A in the proliferation of breast cancer cells by interfering with PKD1-mediated MEK/ERK and PI3K/AKT signaling pathways
Jin ZHOU ; Chengjian LI ; Fuli TAN ; Xinying YANG ; Xiaolin ZHANG ; Liang ZHAO
Journal of Pharmaceutical Practice 2020;38(3):241-244
Objective Breast cancer is one of the deadliest malignancies in the world. ebracteolatain A (EA) is a kind of acetylphloroglucinol extracted from ebracteolatain. To explore the specific mechanism of EA inhibiting the proliferation of breast cancer cell MCF-7, so as to provide a new approach for the clinical treatment of breast cancer. Methods EA with different concentrations were added to breast cancer cell MCF-7 to detect changes in PKD1 protein expression. The plasmid with overexpressed PKD1 was constructed and transfected into cells, and the mRNA and protein expression levels of PKD1 were detected by real-time fluorescence quantitative PCR and Western Blot assay. CCK-8 assay was used to detect changes in cell proliferation capacity. Western Blot assay was used to detect the expression level of PKD1 and its related signaling pathways. Results EA inhibited the expression of PKD1 protein in breast cancer cells with a dose-dependent manner (P< 0.05). When transfected with the overexpressed plasmid, PKD1 was significantly increased in mRNA and protein levels (P<0.001). At the same time, PKD1 overexpression significantly reversed inhibition of EA on MCF-7 proliferation (P<0.001). It was confirmed by signaling pathway analysis that EA might affect the proliferation ability of breast cancer cells by inhibiting PKD1-mediated MEK/ERK and PI3K/AKT signaling activity (P<0.05). Conclusion EA could inhibit the proliferation of breast cancer cells by regulating PKD1-mediated MEK/ERK and PI3K/AKT signaling pathways.
9.Determination of propylthiouracil in human plasma by UPLC-MS/MS
Qunying LI ; Lin LING ; Chengjian LI ; Jin ZHOU ; TianTian LI ; Liang ZHAO ; Lei LÜ
Journal of Pharmaceutical Practice 2020;38(4):350-353
Objective To establish a method for the determination of propylthiouracil in human plasma by UPLC-MS/MS and provide methodological basis for therapeutic drug monitoring (TDM) and bioequivalence test (BE) in clinical. Methods The chromatographic separation was performed on an Agilent SB-C18 column (4.6 mm×150 mm, 5 μm), the mobile phase was methanol and water containing 0.1% formic acid (80∶20, V/V), isocratic elution. MS condition was optimized in the positive ion detection mode by multiple reaction monitoring (MRM), along with the Agilent JetStream electrospray source interface (AJS-ESI). The precursors to the product ion transitions were m/z 171.1→112.1 for propylthiouracil and m/z 176.1→117.0 for the internal standard (IS). Results The calibration curve was linear in the range of 10−5 000 ng/ml for propylthiouracil in human plasma, r=0.999 3. The intra-day and inter-day precision and accuracy were good (RSD<10%,RE<±10%). The matrix effect of different concentrations was less than 110% and the coefficient of variation was less than 5%. The average recovery of different concentrations was 101.60%−113.56%, which conformed with the requirement of methodological validation. Conclusion The method is rapid, sensitive and accurate, which can be used for the determination of propylthiouracil in human plasma.
10.Guideline for the diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients (version 2023)
Yuan XIONG ; Bobin MI ; Chenchen YAN ; Hui LI ; Wu ZHOU ; Yun SUN ; Tian XIA ; Faqi CAO ; Zhiyong HOU ; Tengbo YU ; Aixi YU ; Meng ZHAO ; Zhao XIE ; Jinmin ZHAO ; Xinbao WU ; Xieyuan JIANG ; Bin YU ; Dianying ZHANG ; Dankai WU ; Guangyao LIU ; Guodong LIU ; Qikai HUA ; Mengfei LIU ; Yiqiang HU ; Peng CHENG ; Hang XUE ; Li LU ; Xiangyu CHU ; Liangcong HU ; Lang CHEN ; Kangkang ZHA ; Chuanlu LIN ; Chengyan YU ; Ranyang TAO ; Ze LIN ; Xudong XIE ; Yanjiu HAN ; Xiaodong GUO ; Zhewei YE ; Qisheng ZHOU ; Yong LIU ; Junwen WANG ; Ping XIA ; Biao CHE ; Bing HU ; Chengjian HE ; Guanglin WANG ; Dongliang WANG ; Fengfei LIN ; Jiangdong NI ; Aiguo WANG ; Dehao FU ; Shiwu DONG ; Lin CHEN ; Xinzhong XU ; Jiacan SU ; Peifu TANG ; Baoguo JIANG ; Yingze ZHANG ; Xiaobing FU ; Guohui LIU
Chinese Journal of Trauma 2023;39(6):481-493
Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.