2.Alginate/chitosan microspheres containing cationic beta-cyclodextrin polymer/insulin complex
Lan HUANG ; Jianyu XIN ; Nan ZHANG ; Jianshu LI
Chinese Journal of Tissue Engineering Research 2007;0(08):-
BACKGROUND: Protein drugs such as insulin are usually used in the form of injection due to its low oral administration bioavailability. For the purpose of higher bioavailability, the common method is to encapsulate the drug into algitnate/chitosan microspheres, which introduced polysaccharides to accelerate biological absorption or reduce demolition of protease. OBJECTIVE: To construct alginate/chitosan microspheres containing cationic?-cyclodextrin polymer (CP?CD) /insulin complex for insulin oral delivery system, in addition, to inspect the influence of the charge density of cationic ?-cyclodextrin polymers on insulin release performance in vitro. DESIGN, TIME AND SETTING: The contrast observation experiment was performed at the Laboratory of Department of Polymer Science and Engineering, Sichuan University from November 2006 to July 2008. MATERIALS: ?-cyclodextrin, epichlorohydrin was supplied by Tianjin Bodi Chemical Industry Co., Ltd. Choline chloride and alginate was provided by Kelong Reagent Company, chitosan was purchased from Golden-shell Biochemical Co., Ltd. And insulin was produced by Wanbang Biochemical Pharmaceutical Company. METHODS: Cationic ?-cyclodextrin polymer was synthesized through a one-step polymerization of ?-cyclodextrin, epichlorohydrin and choline chloride. It was combined with insulin and then encapsulated into alginate/chitosan microspheres. In vitro insulin release behavior of the microsphere was studied in simulative gastrointestinal fluid. MAIN OUTCOME MEASURES: The entrapment efficiency, as well as insulin release in simulative gastrointestinal fluid. RESULTS: The charge density of CP?CD had a great impact on the insulin association efficiency of microspheres. The maximum association efficiency could reach (76.3?1.5)%. It showed that cumulative insulin release of microspheres containing CP?CD was less than that of the control group in simulated gastric fluid. The insulin released was 5.8 IU in simulated intestinal fluid, which was higher than the control group. CONCLUSION: Alginate/chitosan microsphere containing CP?CD/insulin complex is a promising system for improving insulin oral delivery efficiency.
3.The inhibitory effect of decorin on proliferation of rabbit epithelial cell
Jian-nan, XIANG ; Gui-lan, ZHANG ; Hai-jiang, ZHANG ; Guo-hua, WANG ; Ming, HUO
Chinese Journal of Experimental Ophthalmology 2012;30(1):41-45
BackgroundResearches found that the posterior capsular opacification (PCO) after lensextraction is associated with the elevation of the transforming growth factor-β(TGF-β).To seek the drug for inhibitingproliferation of lens epithelial cells (LECs) is crucial in the treatment and prevention of PCO.ObjectiveThisstudy was to investigate the preventing effects of decorin on the proliferation of LECs.MethodsRabbit LECs wascultured and passaged.The LECs in growth phase were incubated in 96 well plate at the density of 8×106/L.Decorinwith the concentrations 0.1,1.0,10.0 mg/L was added into the medium for 24,48 and 72 hours respectively.0.1%DMSO was used at the same way as positive control group,and the regular cultured cells worked as blank controlgroup.The inhibitory rates of different concentrations of decorin on the growth of LECs were detected by MTT at 24,48and 72 hours after addition of decorin.The percentage of LECs in different cell cycles in various groups was assayedusing flow cytometry.TGF-β level in medium suspension was detected using ELISA.The expression of TGF-β mRNA in LECs was checked by RT-PCR,and α-SMA expression in LECs was determined using immunochemistry.Results ELISA assay showed a statistical difference in the TGF-β levels of different groups (F=39.24,P=0.03 ).The TGF-β levels in 1.0,10.0 mg/L decorin groups were significantly decreased in comparison with blank control group (P<0.01) and 0.1 mg/L decorin group (P<0.05 ).The inhibitory rates of decorin in the concentrations of ≥ 1.0 mg/L on the growth of LECs were higher than the blank control group,and those in various concentrations of decorin groups were considerably lower in 24 hours compared with 48 and 72 hours ( P<0.05 ) and so was the 48 hours compared with 72 hours (P<0.05 ).The percentages of LECs in G0/G1 phase were ascent in 0.1,1.0 and 10.0 mg/L decorin groups in comparison with G2/M and S phase (P<0.05).Immunochemistry revealed the weak expression of α-SMA in various decorin groups in comparison with control group. Conclusions Decorin can effectively inhibit LECs growth and induce LECs apoptosis in concentration- and time-dependent manner.It is suggested that decorin can be used in the prevention and treatment of after cataract.
4.The Application of fMRI in brain surgery for the lesions in or around the Broca's area of brain
Nan WU ; Bing XIE ; Jiuquan ZHANG ; Chuan LAN ; Hanjian DU ; Guocai WU ; Jian WANG ; Hua FENG
Chinese Journal of Nervous and Mental Diseases 2010;36(1):34-38
Objective To evaluate the value of fMRI guided brain surgery for the lesions in or around Broca's area.Methods Forty-three patients with lesions in or adjacent to the Broca's area were studied.fMRI imaging was obtained by BOLD technique with the tasks of reciting.Fiber tract imaging of white matter was obtained by DTI technique.All functional imaging and anatomic imaging were transferred to neuronavigation system.The technique of direct cortical stimulation was used to validate the language cortex in fMRI.The lesions were resected in microscope.Results Broca's area activation was detected in 38 cases..The distance between the fMRI peak and direct cortical stimulation was rated as overlapping (<1 cm diatance) in 25 cases and neighbouring (<2 cm diatance) in 11 cases.Total lesion resection was achieved in 17 cases, subtotal resection in 14 cases, and partial resection in 12 cases.Postoperative neurological functions were improved in 8 cases, unchanged in 31 cases, and temporary worsen in 4 cases.Conclusions The identification of the Broca's area by reciting task in fMRI is sensitive and precise.The fMRI is helpful to decrease the side effect injury in the brain surgery for the lesions in or around the Broca's area.
5.Cloning and Expression Analysis of two Endosperm Promoters in Rice
Yong-Lan CUI ; Xiao-Li ZHONG ; Yong-Ming ZHANG ; Zhong-Nan YANG ;
China Biotechnology 2006;0(08):-
The cloning of promoter is important for studying the genetic engineering and the regulation of gene expression in plants. Two promoters Os772 and Os359, which are predicted to be highly expressed in the endosperm of rice from the EST database were cloned. After construction of the Os772∶∶GUS and Os359∶∶GUS expression vectors, they were transformed into rice. X-Gluc staining of transgenic plants showed that Os772 and Os359 can promote GUS gene expression in matured endosperm but not in root, stem, leaf and flower. This result indicates Os772 and Os359 are two rice endosperm-specific promoters.
6.Research on ginsenosides composition in wild ginseng leaves in different growing years.
Lan-lan ZHANG ; Wen-yuan GAO ; Zhao-hui SONG ; Nan CAI ; Shui-ping ZHOU
China Journal of Chinese Materia Medica 2012;37(17):2530-2533
OBJECTIVETo analyze ginsenosides composition in wild ginseng leaves with different growth years.
METHODThe analysis was performed on Acquity UPLC BEH Shield RP18 (2.1 mm x 100 mm, 1.7 microm) column, the mobile phase was acetonitrile-0.05% formic acid solution in gradient elution mode. The detection wavelength was at 203 nm. The flow rate was 0.4 mL x min(-1) and column temperature was set at 30 degrees C.
RESULTThirteen ginsenosides were determined by the established UPLC method. In 5-17th growth year ginseng leaf samples cultivated simulating wild conditions, the contents of ginsenosides in the 14th year have the highest content.
CONCLUSIONThe established method is simple, accurate and reliable, can be used in ginsenosides determination and fingerprint research of Panax crude drug. The result provides reliable data for the accumulation of ginsenosides and sustainable utilization of ginseng resources.
Drugs, Chinese Herbal ; analysis ; Ginsenosides ; analysis ; Panax ; chemistry ; growth & development ; Plant Leaves ; chemistry ; growth & development ; Time Factors
7.Reconstruction of embryo using an improved nuclear transfer method.
Ke-Liang WU ; Yong-Xiang SHI ; Zeng-Liang BAI ; Hai-Bin TIAN ; Nan ZHANG ; Lan-Lan LIU ; Chang-Bin LIU
Chinese Journal of Biotechnology 2007;23(1):161-165
Previous methods used for nuclear transplantation were further investigated to develop a method that was both easy to carryout and did not require any special apparatus, such as Piezoimpact or Spindle-View. Following the puncture of zona pellucida with two holes by injection pipette that contained donor nuclei or cells, the injection pipette was pulled back to the perivitelline space while the negative pressure was increased in the holding pipette until the polar body and karyoplasm were wiped off completely. Then a reconstructed embryo was completed by the direct injection of the donor nucleus or cell without pulling out the injection pipette. 200 oocytes were manipulated using this method and it cost about 40 seconds with nucleus injection and about 30 seconds with cell injection to complete a reconstructed embryo. The success rates were 62.6% and 86. 0%, respectively, and enucleation rate was about 73.3% validated by Hoechst 33342. Using this method, the nucleus was completely eliminated and another was injected using the microscope and micromanipulator. Moreover, the efficiency of nuclear transplantation and survival rate of reconstructed embryos were greatly improved. Furthermore, it is very easy to manipulate and popularize in practice.
Animals
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Cell Culture Techniques
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methods
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Cell Nucleus
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metabolism
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Cells, Cultured
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Cloning, Organism
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methods
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Embryo, Mammalian
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cytology
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metabolism
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Embryonic Development
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Female
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Inbred DBA
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Mice, Inbred Strains
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Nuclear Transfer Techniques
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Oocytes
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cytology
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metabolism
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Zona Pellucida
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metabolism
8.Exploring the effect of acupuncture plus mild hypothermia on miRNA-204 and its target gene expressions in CIRI rat brain tissues based on MAPK signal pathway
Yu-Lan TANG ; Mai-Lan LIU ; Jian LUO ; Nan LI ; Guo-Shan ZHANG ; Jie YU ; Qian-Yun YANG ; Yan-Ping YANG
Journal of Acupuncture and Tuina Science 2021;19(5):338-344
Objective: To explore the protective mechanism of acupuncture plus mild hypothermia for cerebral ischemia-reperfusion injury (CIRI) by observing the effects of acupuncture plus mild hypothermia on miRNA-204 and its target gene expressions in CIRI rat brain tissues. Methods: Sixty Sprague-Dawley rats were divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group according to the random number table method (n=10). Except for the blank control group and the sham operation group, rats in the other 4 groups received CIRI modeling. After the model was successfully established, rats in the blank control group were bred routinely for 72 h without any interventions; rats in the sham operation group and the model group were bred routinely for 72 h, and only received binding without other interventions after surgery; rats in the acupuncture group were bred routinely for 72 h, and received acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 26) after binding; rats in the mild hypothermia group were bred routinely for 72 h, and received mild hypothermia intervention for 72 h after binding; rats in the acupuncture plus mild hypothermia group were bred routinely for 72 h, followed by receiving acupuncture as in the acupuncture group and mild hypothermia therapy as in the mild hypothermia group after binding. The neurological impairment score, cerebral infarction area ratio, the expressions of miRNA-204 and its target genes including Map3k8, Ntrk2 and Ppp3r1 in the ischemic hippocampus of each group were observed after 72 h of intervention. Results: Before intervention, compared with the blank control group and the sham operation group, the neurological impairment scores and the infarction area ratios of the modelled rats were statistically significantly increased (all P<0.01), indicating that the model was successful. After intervention, compared with the model group, the neurological impairment scores of the three intervention groups were significantly reduced (all P<0.01); compared with the acupuncture group and the mild hypothermia group, the infarction area ratio in the acupuncture plus mild hypothermia group was significantly reduced (both P<0.01); compared with the model group, the three intervention groups showed significant inhibition of miRNA-204 expression in brain tissues (all P<0.05), which was most significant in the acupuncture plus mild hypothermia group (P<0.01); compared with the acupuncture group and the mild hypothermia group, the Map3k8 expression in the acupuncture plus mild hypothermia group was significantly increased (both P<0.01), but there were no significant differences in Ntrk2 and Ppp3r1 expressions between groups (all P>0.05). Conclusion: Acupuncture, mild hypothermia, and acupuncture plus mild hypothermia reduced the neurological impairment score and the cerebral infarction area in CIRI rats, while acupuncture plus mild hypothermia showed the most significant effect. In regulating miRNA-204 target gene expressions, acupuncture plus mild hypothermia showed the same effect on Ntrk2 and Ppp3r1 expressions, while better effect on Map3k8 expression compared with either acupuncture or hypothermia.
9.Effect of kebimin decoction in treating allergic rhinitis and on blood levels of nitric oxide and superoxide dismutase.
Shi-fu ZHANG ; Li-lan CHENG ; Di-nan HUANG
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(2):118-120
OBJECTIVETo observe the therapeutic effect of Kebimin decoction (KD) on allergic rhinitis (AR) and its effect on blood levels of nitric oxide (NO) and superoxide dismutase (SOD) activity.
METHODSEighty-two AR patients were randomly divided into two groups, the treated group and the control group, 41 in each group. To the treated group, KD was given one dose per day for 10 days as one therapeutic course and 1-3 courses were given successively. The control group was treated with Xinfang Rhinitis capsule for 30 days. Blood levels were determined and compared before and after treatment.
RESULTSThe total effective rate in the treated group was 93%, which was better than that in the control group (51%), the difference was significant (chi 2 = 17.704, P < 0.01). Serum level of NO was higher and that of SOD activity was lower in the AR patients than that in healthy persons (P < 0.01), KD could significantly lower the former and increase the latter (P < 0.01).
CONCLUSIONThe therapeutic effect of KD in treating AR was significant, its mechanism might be related with the lowering of NO and increasing of SOD activity in serum, as well as the scavenging of oxygen free radical.
Adolescent ; Adult ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Free Radical Scavengers ; therapeutic use ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Phytotherapy ; Rhinitis, Allergic, Perennial ; blood ; drug therapy ; Superoxide Dismutase ; blood
10.Determination of six C-Glycoside flavones and antitumor activity of water-soluble total flavonoids from Isodon lophanthoides var. gerardianus.
Yang ZHANG ; Hai-ming TANG ; Ai LI ; Lan-fang XU ; Jian-nan CHEN ; Song HUANG ; Lian HE
China Journal of Chinese Materia Medica 2015;40(8):1543-1547
This research established an HPLC method for determination of six C-Glycoside flavones of warer-soluble total flavonoids from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, and studied the antitumor activity of the warer-soluble total flavonoids. The HPLC system consisted of Kromasil 100-5 C18 (4.6 mm x 250 mm, 5 microm) column and a solution system of methanol, acetonitrile and 0.5% formic acid gradient elution at a flow rate of 0. 8 mL x min(-1) and the wavelength of detector was at 334 nm. The column temperature was 25 degrees C. The antitumor activity of water-soluble flavonoids was assayed using HepG2 cell as the tested cell. The linear ranges of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabinosylapigenin were 0.25-2.53, 0.12-1.20, 0.37-3.69, 0.16-1.63, 0.19-1.92, 0.14-1.42 microg, respectively. The average recoveries (n = 6) were 99.6% (RSD 0.87%), 100.2% (RSD 2.0%), 99.6% (RSD 1.8%), 97.9% (RSD 1.5%), 98.8% (RSD 1.2%), 98.6% (RSD 1.2%), respectively. After exposure in 24, 48, 72 h, the total flavonoids showed inhibitory effect on the proliferation of HepG2 cells with IC50 as the evaluation index, the IC50 values of 1.89, 1.71, 1.51 g x L(-1), respectively. The method is quick, simple and accurate with good re- producibility, and can be used for determination of vicenin II, vicenin III, isoschaftoside, schaftoside, vitexin, 6, 8-di-C-a-L-arabino- sylapigenin in the warer-soluble total flavonoids from L lophanthoides var. gerardianus. The warer-soluble total flavonoids from L lophanthoides have inhibitory effect on the proliferation of HepG2 cells.
Antineoplastic Agents, Phytogenic
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analysis
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pharmacology
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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analysis
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pharmacology
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Flavones
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analysis
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pharmacology
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Humans
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Isodon
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chemistry
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Monosaccharides
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analysis
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pharmacology