1.Therapeutic efficacy of 18 α-Diammonium glycyrrhizinate phosphatidylcholine complex in patients with chronic virus hepatitis
Hong ZHAO ; Yong CHEN ; Jiazhang XU ; Jun WANG ; Chongwen SI
Clinical Medicine of China 2008;24(8):770-772
Objective To investigate the therapeutic effieacy and safety of 18 α-Diammonium glycyrrhizinate phosphatidylcholine complex (DGPC) in patients with chronic hepatitis B and or C with elevated aminotransferase. Methods 55 patients with chronic hepatitis B and or C, with serum alanine aminotransferase (ALT) of 2 to 10 times the upper limit of normal were randomly assigned to receive DGPC or Diammonium glyeyrrhizinate (DG) for 12 weeks. Then they were followed up for an additional 4 weeks. From week 1 to 10, DGPC or DG was given as 150 nag,three times a day (TID). At the 11th week,the drug was given as 100 mg,TID. Then 50 mg,TID for the 12th week. Results ALT was markedly decreased after receiving DGPC 4,8,12 weeks (P=0.00). ALT normalization rate at the end of therapy was similar (38.5% vs 34.5% ,P =0.76). Drug-related adverse events were similar. Conclusion DGPC can rapidly and safely decrease aminotransferase in patients with chronic viurs hepatitis.
3.BLG gene knockout and hLF gene knock-in at BLG locus in goat by TALENs.
Shaozheng SONG ; Mengmin ZHU ; Yuguo YUAN ; Yao RONG ; Sheng XU ; Si CHEN ; Junyan MEI ; Yong CHENG
Chinese Journal of Biotechnology 2016;32(3):329-338
To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals
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Animals, Genetically Modified
;
genetics
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Female
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Fibroblasts
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Gene Knock-In Techniques
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Gene Knockout Techniques
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Goats
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genetics
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Humans
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Lactoferrin
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genetics
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Lactoglobulins
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genetics
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Milk
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chemistry
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Nuclear Transfer Techniques
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Plasmids
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Pregnancy
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Transfection
4.Fluorescence imaging of APP in Alzheimer's disease with quantum dot or Cy3: a comparative study
Li FENG ; Shuyu LI ; Bo XIAO ; Si CHEN ; Renkai LIU ; Yong ZHANG
Journal of Central South University(Medical Sciences) 2010;35(9):903-909
Objective To compare the fluorescence intensity and duration of qdots streptavidin conjugate (QDs-SA) with Cy3 as the molecular probe of β amyloid precursor protein (APP), and to provide evidence for early molecular imaging and diagnosis of Alzheimer's dissease (AD). Methods With the help of laser scanning confocal microscope and flow cytometry, the flurescence probe based on the QDs-SA was used to detect APP in HEK293 cells stably transfected pcDNA3.1/APP, and to compare with conventional fluroimmunoassay Cy3. Results The immunofluorescence staining detection indicated APP expression was mainly located in the plasma membrane. The mean fluorescence intensity of QDs-SA (34.2336±4.2455) was greater than that of Cy3 (21.6023±3.0102)under the confocal fluorescence microscope (P<0.05). After persistent exciting for 12 min, the fluorescence intensity of APP stained by QDs-SA decreased by 27.87%. The other stained by Cy3 decreased by 79.60%. The positive rate of APP staining had no significant difference between the QDs-SA(54.4700±3.4433)% and Cy3 (54.3800±8.5229)% by flow cytometry, but the mean fluorescence intensity had statistical significance(P<0.05). The QDs-SA (1 045.4167±47.3623) was significantly higher than the mean fluorescence intensity of Cy3 (658.5467±55.0591). Conclusion QDs-SA fluorescence probes can effectively recognize APP and are sensitive and exceptionally photostable, suggesting that QDs-SA fluorescence probes could be a potential method in APP detection and offer a novel way for the diagnosis of Alzheimer's disease.
5.Simultaneous determination of clevidipine butyrate and its metabolite clevidipine acid in dog blood by liquid chromatography-tandem mass spectrometry.
Huihui WEI ; Yuan GU ; Yanping LIU ; Guangli WEI ; Yong CHEN ; Changxiao LIU ; Duanyun SI
Acta Pharmaceutica Sinica 2015;50(10):1290-6
A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
6.The findings of bronchial artery change in lung cancer with 16-slice CT
Qing-Si ZENG ; Yong-Fu CHEN ; Xiao-Mei WU ; Ren-Li CEN ; Chao-Liang ZHANG ;
Chinese Journal of Radiology 2001;0(09):-
Objective To evaluate the difference of internal diameter of bronchial artery in big lung cancer,small lung cancer,and normal lung with multiple slice CT.Methods MSCT angiographies of 44 patients with lung cancer confirmed by pathology were retrospectively analyzed,and 29 patients were with big lung cancer(≥3 cm)and 15 patients with small lung cancer(
7.Role of diffusion weighted magnetic resonance imaging and spectroscopy in the diagnosis and follow-up of hepatolenticular degeneration.
Peng XUE ; Yong LIU ; Xiu-hua MA ; Yong CHEN ; Si-jia ZHANG
Acta Academiae Medicinae Sinicae 2012;34(5):497-502
OBJECTIVETo explore the changes of different brain metabolites during hepatolenticular degeneration using diffusion weighted magnetic resonance imaging (DWI) and magnetic resonance spectroscopy (MRS) in patients with hepatolenticular degeneration and study the correlation of apparent diffusion coefficient(ADC) values and MRS with the different pathological changes.
METHODSTotally 53 patients with hepatolenticular degeneration were enrolled in this study and divided into DWI high-signal group (n=31) and DWI low-signal group (n=22). Magnetic resonance scan, DWI, and spectroscopy were performed before treatment and 4 months after treatment. The changes of ADC value, N-acetyl aspartate (NAA)/creatine (Cr) ratio, and choline (Cho)/Cr ratio were recorded.
RESULTSBefore treatment, the NAA/Cr ratio was significantly higher in the DWI high-signal group than in DWI low-signal group (P=0.002), whereas ADC value and NAA/Cr ratio were significantly lower (P=0.004, P=0.014, respectively). After treatment, the NAA/Cr ratio was still significantly higher in the DWI high-signal group (P=0.036), while the differences of ADC value and Cho/Cr ratio showed no statistical deference (P>0.05). In the DWI high-signal group, the ADC value and NAA/Cr ratio were significantly elevated after treatment (P=0.006, P=0.008), whereas the Cho/Cr ratio showed no significant change (P>0.05). In the DWI low signal group, NAA/Cr ratio was significantly increased after treatment (P=0.015), while the ADC value and Cho/Cr ratio showed no significant change (P>0.05).
CONCLUSIONSDWI combined MRS imaging can be used to evaluate the microscopic structure and metabolic changes during copper deposition and thus, compared with the conventional magnetic resonance imaging provide more information on metabolism. Therefore, they can be useful tools in the early diagnosis and efficacy evaluation of hepatolenticular degeneration.
Adolescent ; Child ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Hepatolenticular Degeneration ; diagnosis ; Humans ; Magnetic Resonance Spectroscopy ; methods ; Male ; Young Adult
8.Preliminary pharmaphylogenetic study on Ranunculaceae.
Yong PENG ; Si-bao CHEN ; Shi-lin CHEN ; Pei-gen XIAO
China Journal of Chinese Materia Medica 2006;31(13):1124-1128
OBJECTIVETo deal with the correlation between phylogeny, chemical constituents and pharmaceutical aspects of Ranunculaceae, namely a pharmaphylogenic study of this taxon.
METHODBased on chemical, pharmaceutical (both ethnopharmacologic and pharmacological) information, linking with different plant systems of Ranunculaceae.
RESULTChemical constituents of this taxon included several natural groups: benzylisoquinoline alkaloid, ranunculin, triterpenoid glycoside and diterpene alkaloid etc. Ranunculin and magoflorine were found to present simultaneously in some plants of this taxon.
CONCLUSIONCombining with therapeutic information, pharmaphylogenic research were in accordance with the phylogenetic system presented by Tamura that Ranunculaceae was proposed to be divided into six sub-families: Helleboroideae, Ranunculoideae, Cimicfugoideae, Isopyroideae, Thalictroideae and Coptidoideae. Results also supported the establishment of Cimicifugoideae.
Benzylisoquinolines ; isolation & purification ; Cimicifuga ; chemistry ; Diterpenes ; isolation & purification ; Furans ; isolation & purification ; Helleborus ; chemistry ; Methylglycosides ; isolation & purification ; Pharmacognosy ; classification ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; Ranunculaceae ; chemistry ; classification ; Triterpenes ; isolation & purification
9.Simultaneous determination of clevidipine butyrate and its metabolite clevidipine acid in dog blood by liquid chromatography-tandem mass spectrometry.
Hui-hui WEI ; Yuan GU ; Yan-ping LIU ; Guang-li WEI ; Yong CHEN ; Chang-xiao LIU ; Duan-yun SI
Acta Pharmaceutica Sinica 2015;50(10):1290-1296
A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals
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Butyrates
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blood
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pharmacokinetics
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Calibration
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Chromatography, Liquid
;
Dogs
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Infusions, Intravenous
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Pyridines
;
blood
;
pharmacokinetics
;
Tandem Mass Spectrometry
10.Research progress of skin-derived precursor cells.
Ruo-Si CHEN ; Yong MIAO ; Zhi-Qi HU
Journal of Southern Medical University 2017;37(3):420-422
As a novel population of neural crest-origin precursor cells, skin-derived precursor cells (SKPs) can be isolated from both embryonic and adult dermis. These cells have important values for research and potential clinical application in wound healing, organ regeneration and disease treatment for advantages in the abundance of cell sources, accessibility, potential of multipotent differentiation, and absence of ethical concerns. Here we review the developmental and anatomical origins of SKPs and their potential application in regenerative medicine. SKPs originate from the embryonic neural crest, and their sources may vary in different areas of the body. SKPs are widely found in the dermis, especially in the dermal papilla (DP), which was known as a niche of SKPs. The multipotent SKPs can used for autologous transplantation and are of vital importance in tissue repair.