2.Constructing a lentiviral vector overexpressing indoleamine 2,3-dioxygenase
Jigang HE ; Hongrong LI ; Longsheng GUI ; Yongwu LI ; Dan YAN
Chinese Journal of Tissue Engineering Research 2015;(36):5859-5864
BACKGROUND:Immune rejections after organ transplantation or serious adverse reactions due to immunosuppressive drugs show a lack of effective treatments and poor therapeutic outcomes. Therefore, we try to find an effective immune suprresion method in combination of the latest immunomodulatory achievements. OBJECTIVE:To construct a lentiviral vector overexpressing indoleamine 2,3-dioxygenase (IDO). METHODS:(1) The IDO gene that was successful y contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cel s with 80%confluence were co-cultured with 5'LTR and 3'LTR, basic elements of lentiviral packaging auxiliary components, including Psi, cPPT, 3FLAG, TetR, IRES, WRPE, TetIIP, Ubiquitin Promoter, SV40 origin and HIV. RESULTS AND CONCLUSION:Western blot assay showed that in 10 g/L agarose gel electrophoresis, there was a target fragment at Mr 48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cel s. These findings suggest that IDO fusion gene has been successful y reorganized in the lentiviral packaging plasmids.
3.Preliminary study on efficacy and mechanism of Atractylodes Macrocephelae Rhizoma extracts in metabolic hyperlipidemia rats.
Qi-jing TANG ; Su-hong CHEN ; Dan-dan PAN ; Bo LI ; Gui-yan LV
China Journal of Chinese Materia Medica 2015;40(9):1803-1807
Hyperlipidemia is a major factor causing coronary heart disease and atherosclerosis. The high-density lipoprotein cholesterol (HDL-C) is a major indicator for measuring lipid levels. However, there is no an effective medicine that can obviously increase HDL-C at present. According to previous laboratory studies, atractylodes macrocephalae extracts could significantly increase HDL-C level. In this study, the metabolic hyperlipidemia rat model was established by feeding high-sugar and fat diets and alcohol-drinking to explore the effect and mechanism of atractylodes macrocephalae extracts on hyperlipidemia rats. According to the findingins, different doses of atractylodes macrocephalae extracts could reduce the levels of TC, TG, LDL-C, ACAT and increase the contents of LCAT, HDL-C. Particularly, the atractylodes macrocephalae extracts (100 mg · kg(-1) group showed increase in HDL-C by about 50% and significant declines in HMG-CoA reductase, TC, TG. In conclusion, Atractylodes Macrocephelae Rhizoma extracts could effectively regulate the dyslipidemia of hyperlipidemia rats, especially on HDL-C. Its mechanism may be related to reduction in cholesterol synthesis by inhibiting HMG-CoA reductase in livers and increase in lipid metabolism and transport by regulating LCAT and ACAT levels.
Acyl Coenzyme A
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antagonists & inhibitors
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genetics
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metabolism
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Animals
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Atractylodes
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chemistry
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Drugs, Chinese Herbal
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administration & dosage
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Humans
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Hydroxymethylglutaryl-CoA Reductase Inhibitors
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administration & dosage
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Hyperlipidemias
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drug therapy
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enzymology
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metabolism
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Lipoproteins, HDL
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metabolism
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Male
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Rats
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Rats, Sprague-Dawley
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Rhizome
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chemistry
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Triglycerides
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metabolism
4.The cervical aortic arch in 2 cases
Jigang HE ; Hongrong LI ; Longsheng GUI ; Yongwu LI ; Dan YAN ; Ping WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2016;32(6):332-333
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5.Homing ability of the SCA-1+/CD45+/CD31+ subgroup of bone marrow mesenchymal stem cells
Jigang HE ; Hongrong LI ; Longsheng GUI ; Yongwu LI ; Dan YAN ; Ping WANG
Chinese Journal of Tissue Engineering Research 2015;(41):6572-6578
BACKGROUND:Since the FDA was the first to approve autologous bone marrow stem cel transplantation for treatment of myocardial infarction in 2003, there has a large number of clinical and basic research reports. However, their conclusions are different and stem cel homing is a key point. OBJECTIVE:To explore the homing abilities of different subgroups of mouse bone marrow mesenchymal stem cels in myocardial regeneration. METHODS:After mouse bone marrow mesenchymal stem cels were detected using a mouse cardiac stem cel surface differentiation antigen, four cel subgroups were separated on the basis of CD45 and CD31. The homing abilities of the four subgroups were assayed in a Transwel chamberin vitro. The different cel subgroups were injected into the model mice suffering from myocardial infarction for 48 hours. The mice were sacrificed at 48 hours, 96 hours, and 7 days after injection; the hearts were taken and analyzed through whole-body imaging and fluorescence intensity detection. RESULTS AND CONCLUSION:The SCA-1+/CD45+/CD31+ subgroup exhibited the strongest homing ability. The whole-body imaging indicated that the fluorescence intensity of SCA-1+/CD45+/CD31+ subgroup was higher than that of the other subgroups at 48 hours, 96 hours and 7 days after stem cel injection. The migration rate of SCA-1+/CD45+/CD31+ subgroup was also the highest. These findings indicate that the homing ability of the SCA-1+/CD45+/CD31+ subgroup of mouse bone marrow mesenchymal stem cels exhibit a homing trend to the damaged myocardial tissue.
6.Inhibitory effects of butyl alcohol extract of Baitouweng decoction on virulence factors of Candida tropicalis.
Gui-ming YAN ; Meng-xiang ZHANG ; Dan XIA ; Ke-qiao LU ; Jing SHAO ; Tian-ming WANG ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(12):2396-2402
OBJECTIVETo investigate the effects of butyl alcohol extract of baitouweng decoction (BAEB) on the fungal cell surface hydrophobicity (CSH), filamentation and biofilm formation of Candida tropicalis.
METHODGradual dilution method was used to determine the MIC. XTT assay was applied to determine the SMIC80. Time-Kill assay was employed to draw the Time-Kill curve. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity. Scanning electron microscopy (SEM) was applied to observe the morphological changes of the biofilm. Confocal laser scanning microscopy (CLSM) was applied to determine the thickness of the biofilm. The quantification real-time PCR (qRT-PCR) was used to detect expression changes of releated genes (UME6, ALST3 and NRG1). result: The MICs of BAEB against C. tropicalis strains are determined as 64-128 mg x L(-1). The SMIC80 s of BAEB against the biofilm of Candida tropicalis strains are determined as 256-512 mg x L(-1). Time-Kill curve results indicate that BAEB has a promise fungicidal effect at 256 and 512 mg x L(-1). SEM results shows that 512 mg x L(-1) BAEB can inhibit the formation of C. tropicalis biofilm on Silicone catheter, and the morphology of biofilm is also affected by BAEB. The thickness of C. tropicalis biofilm is reduced by BAEB according to CLSM results. Furthermore, qRT-PCR results indicate that expression of UME6 and ALST3 are significantly down-regulated by BAEB 256,512 mg x L(-1), and NRG1 is not affected by BAEB.
CONCLUSIONBAEB inhibits effectively the CSH, filamentation and biofilm formation of VVC strains of C. tropicalis.
Antifungal Agents ; chemistry ; pharmacology ; Biofilms ; drug effects ; Candida tropicalis ; drug effects ; genetics ; physiology ; Candidiasis ; microbiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Fungal ; drug effects ; Humans ; Virulence Factors ; genetics ; metabolism
7.Chemical constituents from the roots tuber of Aconitum ouvrardianum
Wang-Yan LIU ; Yong SHEN ; Dan HE ; Cheng-Xin GUO ; Gui-Qiong LI ; Xiao-Yan DUAN ; Jiao XIONG ; Yan-Ping CHEN
Chinese Traditional Patent Medicine 2018;40(2):368-374
AIM To study the chemical constituents from the roots tuber of Aconitum ouvrardianum H..METHODS The ethyl acetate fraction of methy extract from A.ouvrardianum roots tuber was isolated and purified by silica,aluminium oxide column,Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Eighteen compounds were isolated and identified as kongboendine (1),anisoezochasmaconitine (2),lipo-14-O-anisoylbikhaconine (3),franchetine (4),talatisamine (5),chasmanine (6),crassicauline A (7),chasmaconitine (8),14-dehydrotalatisamine (9),lipoindaconitine (10),indaconitine (11),yunaconitine (12),lipoyunaconitine (13),liljestrandisine (14),transconitine B (15),ouvrardiantine (16),atropurpursine (17),8-deacetylyunaconitine (18).CONCLUSION Compounds 1-4,9-10,13,15,17 are isolated from this plant for the first time.
8.The Clinical features of myeloma bone disease..
Si-Dan LI ; Yan XU ; Ya-Fei WANG ; Yao-Zhong ZHAO ; Shan JIANG ; Lu-Gui QIU
Chinese Journal of Hematology 2010;31(4):228-232
OBJECTIVETo analyze the clinical features of patients with newly diagnosed myeloma bone disease (MBD).
METHODClinical features of MBD in two hundred and five patients with newly diagnosed multiple myeloma (MM) were analyzed retrospectively. The relationship between outcome of different grades of MBD patients and their prognosis was compared.
RESULTS(1) Among the 205 patients, one hundred and fifty (72.7%) had bone pain as the first symptom. (2) According to X-ray evaluation, there were 23 (11.3%) of grade 0, 14 (6.9%) grade 1, 23 (11.3%) of grade 2, 68 (33.3%) grade 3 and 76 (37.2%) grade 4. (3) Patients with grade2-4 MBD had significantly higher ECOG performance score, marrow plasmacytes, marrow CD138(+) CD38(+) cell percentage and serum IL-6 level than those with grade 0 - 1 did (P < 0.05). Patients with grade 4 MBD presented with hypocalcemia (P < 0.05) more often than those with grades 0 - 3 diseases did. (4) There was no significant difference in response to initial induction chemotherapy among the five groups (P = 0.642). (5) Univariate analysis demonstrated that the time to progression (TTP) in grade 2 - 4 MBD groups was significantly shorter than that in grade 0 - 1 groups (P = 0.029). (6) Multivariable COX analysis did not indicate lytic bone changes was a independent prognostic factor for OS and TTP.
CONCLUSIONThere is a rather high incidence of MBD in MM patients in China. Patients with extensive X-ray bone lesions have more severe hematologic parameters than those without bone lesions did, and severe bone lesions is an important adverse prognostic factor for TTP.
Bone Diseases ; Bone Marrow ; Humans ; Interleukin-6 ; Multiple Myeloma ; Prognosis ; Retrospective Studies
9.Application of an extended collar incision in neck dissection for differentiated thyroid cancer.
Bin ZHANG ; Dan-gui YAN ; Chang-ming AN ; Zhen-gang XU ; Ping-zhang TANG
Chinese Journal of Oncology 2009;31(3):223-225
OBJECTIVETo explore a cosmetic incision in the neck dissection for differentiated thyroid carcinoma.
METHODSAn extended collar incision was used for neck dissection in 82 consecutive patients with thyroid carcinoma from May 1999 to December 2006. The incision was designed to start as a conventional thyroid collar incision, and then to extend it along the skin crease to the anterior border of trapezium, so to avoid the vertical limb of conventional hockey stick incision. There were 60 females and 22 males in this series, with a median age of 40.5 years (range, 10 to 80 years). Ninety-six procedures of neck dissection were performed in 82 patients, including one radical neck dissection, one type I modified neck dissection, 8 type II modified neck dissections, and 86 type III modified neck dissections.
RESULTSThe average time of anesthesia was 197 minutes. The average dissected lymph nodes were 37.5, with average metastasis in 8.8 nodes. Eight patients (9.8%) developed complications related to neck dissection. The follow-up period in these patients were 1 to 96 months with a median follow-up time of 23 months. Cervical recurrence was found in only one patient (1.2%). Neither death nor distant metastasis was observed in this series.
CONCLUSIONIt is feasible to perform a modified neck dissection for differentiated thyroid cancer through the extended collar incision. The preliminary results show that the above described incision is not only oncologically safe, but also offers a cosmetic benefit for the patient with thyroid carcinoma.
Adenocarcinoma, Follicular ; pathology ; surgery ; Adenocarcinoma, Papillary ; pathology ; surgery ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Humans ; Hypocalcemia ; etiology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; adverse effects ; methods ; Thyroid Neoplasms ; pathology ; surgery ; Young Adult
10.Resistance reversal effect of a novel taxane compound NPB304 and its collaboration with verapamil.
Mei MEI ; Yi ZHANG ; Jin-Hong REN ; Dan XIE ; Yu-Fei JIA ; Jin-Ping HU ; Yan LI ; Jun-Gui DAI ; Xiao-Guang CHEN
Acta Pharmaceutica Sinica 2014;49(9):1279-1288
The tumor multidrug resistance reversal effect of NPB304, a novel taxane, was studied. MTT assay was used to determine the IC50 of chemotherapy drugs. Western blotting assay was applied to analyze the expression of P-glycoprotein (P-gp). The effect of compounds on the P-gp function and P-gp ATPase activity was determined by rhodamine 123 (Rh123) accumulation assay and analysis kit, respectively. Molecular docking was employed to predict the binding force between compounds and P-gp. Transmembrane transport of NPB304 was analyzed using MDCK II and MDR1-MDCK II cell model. NPB304 displayed multidrug resistance reversal effect on KBV cells and MCF-7/paclitaxel cells, NPB304 collaborative with P-glycoprotein (P-gp) inhibitors verapamil enhanced the reversal activity, specifically, 10 μmol x L(-1) verapamil in combination with paclitaxel reversed resistance by 56.5-fold, while combined with NPB304 increased the reversal fold; NPB304 synergistically increased Rh123 accumulation in the resistant cells when combined with verapamil, and NPB304 at 0-1 μmol x L(-1) enhanced the ATPase activity activated by verapamil was observed. NPB304 existed the hydrophobic interactions with the TM regions of P-gp, and the binding force between NPB304 and the A chain of the TM region was stronger. P-gp ATPase activity assay demonstrated NPB304 at lower concentrations (0-1.5 μmol x L(-1)) could activate the P-gp ATPase, playing a role on inhibition of P-gp function. However, NPB304 did not have an obvious feature of P-gp substrate. NPB304 exerted itself and synergy with verapamil activity on reversing tumor resistance via inhibiting the P-gp function.
ATP-Binding Cassette, Sub-Family B, Member 1
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metabolism
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Antineoplastic Agents
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pharmacology
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Biological Transport
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Drug Resistance, Multiple
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Drug Resistance, Neoplasm
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Drug Synergism
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Humans
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MCF-7 Cells
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Rhodamine 123
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Taxoids
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pharmacology
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Verapamil
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pharmacology