2.Levonorgestrel-releasing intrauterine system for treatment of dysmenorrhea associated with adenomyosis
Jie SHENG ; Dan LU ; Jian-Ping ZHANG ; Gui-Fen CUI ;
Chinese Journal of Obstetrics and Gynecology 2001;0(07):-
Objective To evaluate the efficacy and safety of a levonorgestrel-releasing intrauterine system(LNG-IUS)for the treatment of dysmenorrhea associated with adenomyosis.Methods We recruited 48 women with moderate or severe dysmenorrhea associated with adenomyosis.All women were inserted of LNG-IUS into their uterine cavity from days 5-7 of their periods and maintained for 12 months.We compared the visual analogue scale(VAS)scores and verbal rating scale(VRS)scores of their dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up.Results Forty-four women completed the study. There were significant differences between mean VAS and VRS scores changes of dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up,those of dysmenorrhea dropping from 75?13 to 11?11 and 2.3?0.4 to 0.4?0.3,those of dyspareunia dropping from 54?19 to 4?4 and from 1.6?0.8 to 0.2?0.2 respectively.Overall 29 women(66%)were very satisfied or satisfied with the one-year treatment. Conclusion Insertion of LNG-IUS alleviates moderate or severe dysmenorrhea associated with adenomyosis remarkably.
3.Concise Plasmids for Antibiotic Resistance Cassette Recovery and in Vitro Recombination
Da-Wei LIU ; Dan-Dan NIU ; Liang ZHANG ; Gui-Yang SHI ; Zheng-Xiang WANG ;
Microbiology 1992;0(05):-
In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.
4.Clonal heterogeneity in differentiation potential of immortalized human mesenchymal stem cells in vitro and in vivo
Zelin SUN ; Yazhuo ZHANG ; Songbai GUI ; Hongyun WANG ; Meizhen SUN ; Dan LI
Clinical Medicine of China 2009;25(8):785-788
Objective To study the clonal heterogeneity in differentiation potential of immortalized mesenchymal stem cells in vitro and in rive.Methods The monoclonal cell lines were performed with limiting dilution cloning,and were induced to adipocytic,asteogenic and neuronal differentiation in vitro.After transplanted the monoclonal cell lines into SCID mice,the xenotransplants were removed and evaluated by immunohistochemistry.Results From the parental HMSC-TERT,32 single-cell derived clones were established,of which the differentiation properties varied considerably in vitro.The cells grow in different plating densities during expansion in culture:HMSC-TERT-2 expressed more strongly in LCA,GFAP and vimemin;HMSC-TERT-C19 expressed more strongly in keratinose than HMSC-TERT-2,HMSC-IERT-20,and MSC-H;HMSC-TERT-C2 expressed more strongly in actin than HMSC-TERT-2.Conclusions The HMSC-TERT monoclone cells are heterogeneity in differentiation petential in vitro and in vivo,suggesting that standard in vitro culture and in vivo inoculate procedure phy an important role in the clinical application of stem cells.
5.Quantitative synaptic alterations in human brain during normal aging and in patients with Alzheimer disease
Dan XU ; Yazhuo HU ; Qiuping GUI ; Mingwei ZHU ; Honghong ZHANG ; Luning WANG
Chinese Journal of Tissue Engineering Research 2005;9(24):260-262
BACKGROUND: Synaptic density, a key index of structure and function of brain tissues, is related to cognitive function. Synaptic loss occurs during human brain aging and in Alzheimer disease (AD), inducing the changes of synaptic density.OBJECTIVE: To observe quantitative synaptic alterations in human brain and changes of synaptic density in different parts during normal aging so as to compare them with those of AD patients.DESIGN: Sampling survey.SETTING: Senile Neurological Department of General Hospital of Chinese PLA.PARTICIPANTS: Pathological data were selected from General Hospital of Chinese PLA from June 1996 to December 2002. Inclusion criteria: had no major nervous system diseases and neuropathological changes. Brain tissues of 28 corpses in normal aging group, 23 males and 5 females aged 23-100 years with an average of (65±22.8) years, were obtained at autopsy.All corpses were divided into three groups according to their age, namely,adult group (23-55 years old, n=9), senile group (64-72 years old, n=7),and >75 group (76-100 years old, n=12). Cerebral hippocampal samples of other six corpses diagnosed with AD were selected from clinic. The corpses included 5 men and 1 woman aged 76-94 years with an average of (83±7.7) years.METHODS: Response intensity of synaptophysin immunochemistry remained stable after 4-8 hours of death, so brains were obtained at autopsy after 8-72 hours of death and fixed with 4% formalin for at least 6 weeks.In normal aging group, tissues were taken from left superior frontal gyrus,striatal area of left occipital lobe, left putamen (striatum section, including head of caudate nucleus), and left hippocampus (from lateral geniculate body section to medial occipitotemporal gyrus). In AD cases, tissues were taken from left hippocampus of 4 corpses and right hippocampus of other 2. All sections were stained with hematoxylin eosin (HE), toluidine blue and synaptophysin immunostaining (rabbit anti-human synaptophysin polyclonal antibody from Beijing Zhongshan Biotechnology Co., Ltd.). Morphology and distribution of positive objects in synapse immunologic reaction were observed under the light microscope. Relation between absorbance in each region and age was determined with Pearson's coefficient. Differences among groups were analyzed with nonparametric test, and the differences in hippocampal CA3 area between > 75 group and AD group were analyzed with the same test.MAIN OUTCOME MEASURES:① Absorbency of synaptophysin at various sites of normal aging group and correlation with age; ② absorbance value in CA3 area between AD patients and advanced aged normal subjects (>75 years) was compared.RESULTS:All the 34 cerebral samples entered the final analysis.①Synaptophysin-positive granules of various size were scattered through neocortex, putamen and hippocampus, neuronal somata, neuroglia, vessels and white matter. Density was particularly strong over layers Ⅱ and Ⅲ in frontal lobe, and layer ⅣV in occipital lobe. ② Synaptophysin density was negatively correlated with age, which was -0.688 in frontal lobe, -0.592 in occipital lobe, -0.458 in putamen and -0.619 in hippocampal CA2 area,respectively (P = 0.000, 0.001, 0.014, and 0.000). ③ Significant difference in synaptic density in CA3 area was found between AD patients (0.031 3±0.003 0)and normal subjects over the age of 75 (0.040 7±0.005 3) (Z=-2.997, P=0.001)in nonparametric test.CONCLUSION:① Synaptic density was found to decrease in frontal lobe, occipital lobe, CA3 area of hippocampus and putamen with age; the changes had significant correlation with age.② Synaptic density of AD patients was lower than that of normal subjects, and their cognitive hypofunction was related to synaptic loss. ③ All tissues were obtained after 8-72 hours of death and fixed over 6 weeks, which to the greatest extent reduced the effects of tissue autolysis and formalin fixation on the results.
6.Effect of all-trans retinoic acid on the excretion of urine TGF ?_1 and glomerular lesion of diabetic rat
Dan ZHU ; Baosong GUI ; Ganglian YAO ; Dengfeng GAO ; Ning NING ; Hongli ZHANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2004;0(05):-
Objective To investigate the effect of all-trans retinoic acid(atRA) on the urinary TGF?_1 excretion and glomerular lesion of diabetic rats at early stage.Methods SD rats were randomly assigned into 3 groups: atRA treated group,diabetic control group and normal control group.Streptozotocin(STZ)-induced early diabetic male SD rat were used.The atRA treated group were treated with daily subcutaneous injections atRA of 10mg/kg for 7 days(n=6),and then the excretion of urinary protein and TGF?_1 and NO level of plasma,urine and renal tissue were measured,and pathological changes of their kidneys were observed.Results The diabetic control rats showed increased urinary excretion of protein and TGF?_1 and NO level of plasma,urine and renal tissue and deposit of glomerular matrix,while atRA prevented these changes.Conclusion AtRA can prevent the development of diabetic nephropathy,which is relevant with the inhibition of secretion of TGF?_1 and NO.
7.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
8.Effects of Jiji decoction on the cognitive function and oxidative stress in mice with vascular dementia induced by cerebral ischemia/reperfusion.
Feng-xi LIU ; Kai NIU ; Zhi-gang WU ; Gui-ping XUE ; Jian-ming YANG ; Dan-shen ZHANG
Chinese Journal of Applied Physiology 2015;31(2):170-177
OBJECTIVETo determine the effects of Jiji decoction (Traditional Chinese Medicine) on the cognitive function and oxidative stress in mice with vascular dementia (VD) induced by cerebral ischemia/reperfusion.
METHODSThirty-two mice were randomly divided into nonnal group (n = 8), sham group (operation, but no cerebral ischemia/reperfusi6n, n = 8), model group (vascular dementia model induced by cerebral ischemia/reperfusion, n = 8), and Jiji decoction-treated group (vascular dementia model plus treatment with Jiji decoction, n = 8). Fourteen days of treatment after operation, the cognitive behavior was measured in step-through test, spatial probe test and platform test. Afterwards, to assess the levels of oxidative stress, the activity of superoxide dismutase(SOD) and content of malonaldehyde (MDA) in brain of these mice were measured.
RESULTSData from step-through test indicated that the escaping latency of Jiji decoction-treated group was prolonged and the error counts were decreased significantly ( P <0.01) compared with those of model group. Data from spatial probe test indicated that the time of entering darkroom, the time of climbing height and the time of entering bright room in Jiji decoction-treated group were shortened and the counts of climbing height were increased (P < 0.05-0.01) significantly compared with those of model group. Data from platform test showed that the escaping latency of Jiji decoction-treated group was prolonged significantly (P < 0.01) compared with that of model group. Compared with normal and sham group, the activity of SOD was decreased and the content of MDA was increased in model group significantly (P < 0.01). Compared with those of model group, the levels of SOD and MDA in Jiji decoction-treated group were improved significantly (P < 0.01).
CONCLUSIONJiji decoction could improve cognitive function of VD mice. Its mechanism might be related with the inhibition of oxidative stiess in the brain.
Animals ; Brain ; drug effects ; metabolism ; Cerebral Infarction ; physiopathology ; Cognition ; drug effects ; Dementia, Vascular ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Malondialdehyde ; metabolism ; Medicine, Chinese Traditional ; Mice ; Oxidative Stress ; drug effects ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism
9.Effect of osthol on memory impairment of mice in AlCl3-induced acute senile model.
Li-xia SHEN ; Le-qun JIN ; Dan-shen ZHANG ; Gui-ping XUE
Acta Pharmaceutica Sinica 2002;37(3):178-180
AIMTo study the protective effect and mechanism of osthol on learning and memory impairment of mice with acute senile model induced by AlCl3.
METHODSAfter s.c. AlCl3 60 mg.kg-1 for 7 d and i.p. osthol 15 and 7.5 mg.kg-1 for 12 d, using step-through test and step-down test, the effect of osthol on learning and memory was observed and the glutathione peroxidase (GSH-PX) activities in blood and superoxide dismutase (SOD) activities in plasma and cerebrum were measured.
RESULTSOsthol 15 and 7.5 mg.kg-1 significantly improved the capability of memory and enhanced the activities of GSH-PX and SOD in AlCl3 treated mice.
CONCLUSIONOsthol shows protective effect on brain memory impairment of mice in acute senile model induced by AlCl3. Perhaps the mechanism is involved in enhancing the activities of GSH-PX and SOD, clearing away the free radical, protecting the brain neuron from the harm of lipoperoxide.
Acute-Phase Reaction ; Aging ; drug effects ; metabolism ; Aluminum Compounds ; pharmacology ; Animals ; Avoidance Learning ; drug effects ; Brain ; enzymology ; Chlorides ; pharmacology ; Cnidium ; chemistry ; Coumarins ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Glutathione Peroxidase ; blood ; Male ; Memory Disorders ; chemically induced ; enzymology ; prevention & control ; Mice ; Plants, Medicinal ; chemistry ; Random Allocation ; Superoxide Dismutase ; metabolism
10.Prokaryotic Expression and Acetylation Assays of Histone Acetyltransferase PCAF
Wei-Zhong LI ; Dan-Gui ZHANG ; Jun ZENG ; Ge-Fei WANG ; Xiao-Xuan CHEN ; You-Ying CHEN ; Kang-Sheng LI ;
China Biotechnology 2006;0(01):-
P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.