2.Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer
Gen-Shen ZHONG ; Ai-Ping YU ; Ji-De JIN ; Zhong-Hua JIANG ; Zu-Ze WU ;
China Biotechnology 2006;0(02):-
The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.
3.Dihydroartemisinin enhances antitumor effect of 5-fluorouracil against gastric cancer by down-regulating SIRT1 expression
Bin WAN ; bin Heng CAO ; hua Gen YU
Chinese Journal of Pathophysiology 2017;33(12):2195-2201
AIM:To investigate the effect of dihydroartemisinin ( DHA) adjuvant treatment on enhancing the antitumor effect of 5-fluorouracil (5-FU) against gastric cancer .METHODS:The gastric cancer BGC-823 cells were di-vided into control group , DHA group, 5-FU group, 5-FU+DHA group and 5-FU+DHA+SIRT1 plasmid group.The via-bility of BGC-823 cells treated with DHA and 5-FU was measured by MTT assay .The expression of SIRT1 and NADPH ox-idase, activation of caspase-9 and caspase-3, and phosphorylation of ASK1 and JNK in the BGC-823 cells treated with DHA and 5-FU were determined by Western blot .The production of ROS and the apoptosis of the BGC-823 cells treated with DHA and 5-FU were analyzed by flow cytometry .RESULTS:Dihydroartemisinin significantly inhibited the expression of SIRT1 and increased NADPH oxidase protein level (P<0.05).DHA increased the sensitivity of BGC-823 cells to 5-FU, thus decreasing the IC50 of 5-FU to the gastric cancer cells.However, transfection with SIRT1 plasmid decreased the cytotoxicity of DHA and 5-FU co-treatment to the BGC-823 cells.DHA promoted the production of ROS and phosphoryla-tion of ASK1 and JNK induced by 5-FU in the BGC-823 cells ( P<0.05 ) .However , ROS scavenger N-acetylcysteine ( NAC) or JNK specific inhibitor SP600125 inhibited the cell death and activation of caspase-9 and caspase-3 induced by DHA and 5-FU co-treatment (P<0.05).In addition, NAC significantly inhibited the phosphorylation of JNK in the BGC-823 cells co-treated with DHA and 5-FU.However, treatment with SP600125 did not influence the ROS production in the BGC-823 cells, indicating that JNK was the downstream target of ROS pathway .CONCLUSION: Combination of DHA with 5-FU induces caspase-dependent apoptosis in gastric cancer cells through the SIRT 1/NADPH oxidase/ROS/JNK sig-naling pathway .
4.Quantification of pentoxyverine citrate in human plasma by LC-ESI/MS method and its application.
Ke-Hua YU ; Jin-Hua WEN ; Fu-Gen NIE ; Yu-Qing XIONG
Acta Pharmaceutica Sinica 2009;44(12):1402-1405
A rapid and sensitive liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI/ MS) method for quantification of pentoxyverine citrate in human plasma has been developed and applied for the bioequivalence and pharmacokinetics study. After extracted from plasma samples with ethyl acetate, analysis was performed in selected ion monitoring (SIM) mode with a positive electrospray ionization (ESI) interface with a mobile phase consisted of methanol and water (0.4% glacial acetic acid and 4 mmol x L(-1) ammonium acetate, 43 : 57, v/v). The linear concentration range of the calibration curves was 1.0-160.0 ng x mL(-1) for pentoxyverine citrate, inter- and intra-precision (RSD) was less than 12.5%, accuracy (RE) was in +/- 13.5% and absolute recovery was more than 80%. The method was proved simple, rapid, sensitive, specific and suitable for pharmacokinetic and bioequivalence study of Yufenweilin capsule containing pentoxyverine citrate.
Chromatography, Liquid
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methods
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Cyclopentanes
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blood
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pharmacokinetics
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Humans
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Male
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Sensitivity and Specificity
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Spectrometry, Mass, Electrospray Ionization
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methods
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Therapeutic Equivalency
5.Literature survey on botanical origin and clinical application of traditional Tibetan medicine "Shengdeng".
De-Dao CHEN ; Xian-Hua MENG ; Ying-Shan ZHANG ; Gen-Ping CHEN ; Yu-Lan HUANG
China Journal of Chinese Materia Medica 2012;37(20):3147-3149
"Shengdeng" is its Tibetan transliteration referring to many medicines. Tibetan doctors and pharmacists in different areas use different drugs in formulation and clinical application, which are easily confused. In order to grasp the formula and clinical application accurately, we conduct a literature survey on history and current state of botanical origin and clinical application of "Shengdeng", making clear the application of various herbs named "Shengdeng" and providing reference to all Tibetan researchers and clinical workers in formulation and clinical application.
Drug Therapy
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history
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Drugs, Chinese Herbal
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analysis
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history
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therapeutic use
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History, Ancient
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Humans
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Medicine, Tibetan Traditional
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history
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Plants, Medicinal
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chemistry
6.Retrospective analysis of nutritional risk and nutritional treatment options in inflammatory bowel disease
Yu TIAN ; Hua-Hong WANG ; Jun-Xia LI ; Gui-Gen TENG ; Xin-Guang LIU
Parenteral & Enteral Nutrition 2018;25(2):98-101,106
Objective:To understand the nutritional risk in patients with IBD,its related factors and nutritional treatment options.Methods:IBD patients treated in Peking University first hospital from January 2006 to December 2015 were studied.Using the Nutritional risk screening 2002 (NRS2002) nutritional risk assessment of the patients were evaluated.According to the body mass index (BMI),patients were divided into normal BMI group (BMI 18.5 ~ 23.9),low BMI group (BMI < 18.5) and high BMI group (BMI ≥ 24).We analyzed the nutritional risk related factors and compared the difference of nutritional therapy options,regarding the UC and CD patients respectively.Results:A total of 388 patients with IBD were enrolled in the study,with UC 306 and CD 82 patients.The total nutritional risk was 49.5%.Although there was no difference in BMI distribution between UC and CD,CD was more likely to have nutritional risk than UC (CD 64.6%,UC 45.4%,(P =0.002).The nutritional risk of low BMI group was 95.7%.There were no differences in age,sex,and family history in IBD patients for the occurrence of nutritional risk.The more frequently recurrence,severe of disease activity,and the wider rang of disease bring the higher nutritional risk for UC patients.But for CD patients,penetrating type,having a history of surgery and severe of disease activity had higher nutritional risk.Adequate caloric nutrition therapy in patients with CD was 77.4% higher than that of UC 46.8%,(P < 0.001).It was a main principle of our center that UC patients with severe recurrence should not emphasize enteral nutrition and CD patients should first deal with the contraindications before starting enteral nutrition.Conclusions:IBD patients have a high nutritional risk and CD is more obvious than UC,particularly in low BMI patients.The nutritional risk of patients with UC and CD has its own associated factors.It is safe to treat IBD patients with enteral nutrition as long as the indications and contraindications were well controlled.
7.Basic and clinical application progression of invigorating blood and dissolving stasis Chinese medicine in ophthalmology
Ying, WANG ; Jun, PENG ; Gen-Yan, QIN ; Yu, XIANG ; Jia-Qi, LIU ; Xiang-Dong, CHEN ; Ya-Sha, ZHOU ; Qing-Hua, PENG
International Eye Science 2017;17(6):1019-1022
Invigorating blood and dissolving stasis method is a kind of unique therapy of Traditional Chinese Medicine(TCM) treatment, which efficacy has become increasingly prominent in the treatment of ophthalmology.With the further studies of blood stasis and invigorating blood and dissolving stasis therapy, it is widely used in clinical ophthalmology, and get good effects beyond thought, especially when western medicine has no curative effects.It improved the cure rate of fundus oculi disease from the eyelids, conjunctiva, lacrimal sac, vitreous body to the choroid and retina, optic nerve and macula lutea, from surface to fundus, or pathological changes related to inflammation, degeneration, necrosis, atrophy, hyperplasia of fibrous tissue hyperplasia.This paper is aim to explain the definition of invigorating blood and dissolving stasis and make a review of basic research and clinical application about it in several diseases.
8.Relationship between the expression of HBV DNA, HBV cccDNA in human ovary tissues and the HBV intrauterine infection
Min-Min YU ; Xiao-Jun GU ; Yin XIA ; Gen-Ju WANG ; Nai-Ying KAN ; Kai-Hua WU
Chinese Journal of Epidemiology 2013;34(2):178-182
Objective To investigate the relationship between hepatitis B virus (HBV)deoxyribonucleic acid (DNA) and HBV covalently closed circular DNA (cccDNA) in the ovary and HBV intrauterine infection.Methods HBV DNA and HBV cccDNA were assayed in the ovaries of 33 pregnant women who were positive for HBV DNA,tested by Fluorescent quantitative polymerase chain reaction (FQ-PCR).The level of HBV mark (HBVM) and the content of HBV DNA in peripheral blood of infants were measured by chemoluminescence and FQ-PCR methods respectively.Results The overall positive rate for both HBV DNA and HBV cccDNA in ovarian samples was 51.52% (17/33).The rate on intrauterine infection among infants was 12.12% (4/33) and all the 4 infected infants were delivered from mothers with normal hepatic function.When HBV DNA and HBV cccDNA were both positive,the rate of intrauterine infection in infants was significantly higher than those who were with both negative results (P<0.05).Levels of HBV cccDNA and the rate of positive samples were significantly higher in mothers with infants who appeared to have had intrauterine infection than those did not (P<0.01 and <0.05,respectively).Conclusion HBV infection could be discovered in the human ovary and might be transmitted to the filial generation via ovum.
9.Research on HBV DNA inhibition of plasmid acute infection mouse with betulinic acid.
Bing QIAO ; Yue-Qiu GAO ; Man LI ; Shao-Fei WU ; Chao ZHENG ; Shu-Gen JIN ; Hui-Chun WU ; Zhuo YU ; Xue-Hua SUN
China Journal of Chinese Materia Medica 2014;39(6):1097-1100
Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease
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Animals
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Antiviral Agents
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pharmacology
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DNA Replication
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drug effects
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DNA, Viral
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biosynthesis
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Hepatitis B
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blood
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virology
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Hepatitis B Surface Antigens
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blood
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Hepatitis B virus
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drug effects
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genetics
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immunology
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physiology
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Male
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Mice
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Plasmids
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genetics
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Triterpenes
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pharmacology
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Virus Replication
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drug effects
10.Prospective Comparison of Redo Microvascular Decompression and Percutaneous Balloon Compression as Primary Surgery for Recurrent Trigeminal Neuralgia
Jing nan CHEN ; Wen hua YU ; Hang gen DU ; Li JIANG ; Xiao qiao DONG ; Jie CAO
Journal of Korean Neurosurgical Society 2018;61(6):747-752
OBJECTIVE: To prospectively compare facial pain outcomes for patients having either a repeat microvascular decompression (MVD) or percutaneous balloon compression (PBC) as their surgery for trigeminal neuralgia (TN) recurrence.METHODS: Prospective cohort study of 110 patients with TN recurrence who had either redo MVD (n=68) or PBC (n=42) from July 2010 until September 2016. The mean follow-up was 45.6 months.RESULTS: After redo MVD, 65 patients (95.6%) experienced immediate relief of pain. After PBC, 34 patients (81%) were immediately relieved of their neuralgia. After 1 month, the clinical effect of redo MVD was better than PBC (p < 0.01). Patients who had redo MVD more commonly were pain free off medications (93.4% at 1 year, 78.2% at 4 years) compared with the PBC patients (85.1% at 1 year, 59.3% at 4 years). However, mean length of stay was longer (p>0.05). Patients after PBC who occurred developed herpes simplex (35.7%), facial numbness (76.2%), and annoying dysesthesia (21.4%) more frequently compared with patients after redo MVD who occurred developed herpes simplex (14.7%), facial numbness (8.8%), and hypoesthesia (5.9%) (p < 0.05). The symptoms recurred respectively in 15 patients (22.1%) and 19 patients (45.2%) after redo MVD and PBC within the entire 6-year follow-up period.CONCLUSION: For the patients with TN recurrence, redo MVD was a more effective procedure than PBC. The cure rate and immediate relief of pain were better, and the incidence of complications was lower.
Cohort Studies
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Facial Pain
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Follow-Up Studies
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Herpes Simplex
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Humans
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Hypesthesia
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Incidence
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Length of Stay
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Microvascular Decompression Surgery
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Neuralgia
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Paresthesia
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Prospective Studies
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Recurrence
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Trigeminal Neuralgia