1.Preparation and release mechanism of gestodene reservoir-type intravaginal rings.
Chun-Xiao LI ; Yan-Kun WANG ; Mei-Ying NING
Acta Pharmaceutica Sinica 2014;49(3):399-405
This study taking gestodene (GEST) as a model, investigated the factors affecting reservoir-type intravaginal ring (IVR)'s drug release. This paper reported a gestodene intravaginal ring of reservoir design, comprising a gestodene silicone elastomer core encased in a non-medicated silicone sheath, separately manufactured by reaction injection moulding at 80 degrees C and heating vulcanization at 130 degrees C is reported. The test investigated the factors affecting drug release through a single variable method, taking the drug release rates of 21 days as standards. When changing the thickness of the controlling sheath outside, the ratio of the first day of drug release and mean daily release (MDR), named the relatively burst effect, is closing to 1 with the thickness of controlling sheath increasing, while the 1.25 mm sheath corresponding to 1.04 controlled the burst release effectively; a positive correlation (r = 0.992 2) existed between the average drug release (Q/t) and drug loading (A) within a certain range. The C6-165 controlling sheath with high solubility of GEST is easier to achieve controlled release of the drug; GEST crystalline power is more effective to implement controlled release of drugs among difficent states of the drug. A 1/4 fractional segment core gives a relatively burst effect of 1.76, while the 1/1 and 1/2 are 1.93 and 1.87 separately, at the same drug loading, concluding that use of a fractional segment core would allow development of a suitable GEST reservoir IVR. In summary, GEST reservoir-type IVR could be adjusted by the thickness of controlling sheath, the loading of drug, the material properties of controlling sheath, the dispersion state of drug, the additive composition and structure of intravaginal ring, to control the drug release behavior and achieve the desired drug release rate.
Administration, Intravaginal
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Contraceptive Agents, Female
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administration & dosage
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Contraceptive Devices, Female
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Delayed-Action Preparations
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Drug Delivery Systems
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methods
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Norpregnenes
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administration & dosage
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chemistry
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Silicone Elastomers
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chemistry
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Solubility
2.Effect evaluation of skin protection by 3M painless protective membrane after deep venous catheterization
Yali LI ; Yong CUI ; Kun JI ; Mei ZHAO
Chinese Journal of Practical Nursing 2008;24(19):45-46
Objective To discuss the effective measures for skin allergy caused by hyaline mem-brane after deep venous catheterization. Methods Patients (102 eases) were divided into the control group(50 cases) and the test group(52 cases).3M hyaline membrane was patched on the skin directly to fix the catheter after successful puncture in the control group.While in the test group 3M hyaline membrane was patched on the skin after 3M painless protective membrane was sprayed on the skin. Results The incidence of skin allergy in the test group was lower than that of the control group. Conclusions Spray of 3M painless protective membrane before usage of hyaline membrane to fix the catheter could prevent the incidence of skin allergy after deep venous catheterization.
3.Transfection of rabbit bone marrow mesenchymal stem cells with adenovirus vector carrying green fluorescent protein
Rui HUANG ; Yu WANG ; Kun LI ; Jiwen MEI ; Xiaodan JIANG
Chinese Journal of Tissue Engineering Research 2009;13(49):9663-9667
BACKGROUND: It is a new tendency to treat central nervous system injury or tumor therapy using the combination of seed cells and gene therapy.OBJECTIVE: To observe the dose-relationship between transfection and expression of rabbit bone marrow mesenchymal stem cell (BMSCs) with adenovirus vector carrying green fluorescent protein (Ad-GFP), and to study its effects on cell biological properties, in addition, to explore the feasibility of using Ad-GFP vector to construct gene modified BMSCs.DESIGN, TIME AND SETTING: A randomized grouping, contrast observation. The experiment was performed at the Southern Medical University between August 2008 and March 2009.MATERIALS: New Zealand white rabbits, irrespective of genders, weighing 2.0-3.0 kg, were selected.METHODS: BMSCs were separated and cultured in vitro, and then the cell immunophenotypes were detected by flow cytometry.The adenovirus was obtained by packaging 293 cells and was used to transfect BMSCs with various liters (1 ×-10~3-1×10~(10) PFU/mL).Cytometry was used to analyze the transfection efficiency.MAIN OUTCOME MEASURES: Cell morphological changes were detected under an invert microscope. The cell proliferation was detected by CCK8 kits. BMSCs transfected with Ad-GFP were induced differentiating into neuron-like cells by adding of β-mercaptoethanol.RESULTS: The surface markers of 3-6-generation BMSCs were negative to CD34 and CD45, but positive for CD29 and CD44.When the virus titers were 1 ×10~7 PFU/mL, the transfection rate was 55%, which were 85% when the virus titers were 1 ×10~9 and1×10~(10) PFU/mL. However, cell pathology phenomenon occurred when the virus titer was 1 ×10~(10) PFU/mL. The fluorescence was strongest expressed at day 7, and it still can be seen at day 28. The BMSCs trasfected with Ad-GFP could differentiate into neuron-like cells under induction of p-mercaptoethanol, with positive neuron-specific enolase.CONCLUSION: Ad-GFP with suitable titers can infect BMSCs effectively with little influence on the biology property or differentiation function. BMSCs can serve as seeds cell in gene therapy field when utilizing ad-GFP vector system.
5.Autophagy in human skin fibroblast model for photoaging
Qingsong ZHANG ; Mei JU ; Kun CHEN ; Xinyu LI ; Baozhu CHANG ; Heng GU
Chinese Journal of Dermatology 2010;43(8):572-574
Objective To observe the changes of autophagy in human skin fibroblast (HSF) model for photoaging. Methods HSF model for photoaging was established through repeated exposure to ultraviolet B (UVB). Those HSFs receiving no irradiation served as the control. The degree of aging was evaluated by p-galactosidase assay, and autophagy level was detected. Results After repeated exposure to UVB, most pho-toaged HSFs were deformed and distorted, and some of them even died. The percentage of P-galactosidase-positive cells was 50.60% ± 5.04% and 14.58% ± 2.69%, respectively in photoaged and control HSFs (P< 0.01). Significant difference was also observed in the proportion of cytophagosome-positive cells between photoaged and control HSFs (14.91% ± 4.59% vs 68.45% ± 8.25%, P < 0.01). Conclusion The HSF model for photoaging shows obviously abnormal appearance and stagnant growth with increased degree of senescence and decreased autophagy compared with normal control HSFs.
6.Construction of acid-sensitive potassium channel-3 eukaryotic expression plasmid and its express in SH-SY5Y cells.
Lin-yu WEI ; Xin-juan LI ; Yi-wen MEI ; Guo-hong WANG ; Qi WANG ; Dong-liang LI ; Chao-kun LI
Chinese Journal of Applied Physiology 2015;31(3):211-215
OBJECTIVETo construct the acid-sensitive potassium hannel-3(TASK3) eukaryotic expression plasmid and to establish a stable SH-SY5Y cell line expressing enhanced green fluorescent protein (EGFP)-tagged TASK3.
METHODSTASK3 coding region was subcloned into pEGFP-N1 plasmid to construct a recombinant vector alled pEGFP-TASK3. The correct recombinant expressing plasmid was transfected with X-feet transfection reagent to SH-SY5Y cells. The cell line stably expressiing EGFP tagged-TASK3 gene was established by screening with antibiotic G418 and fluorescence microscope. The expression and localization of the EGFP tagged-TASK3 fusion protein was detected by Western blot and confocal microscope. Exposure of the SH-SY5Y cell line expressing stably TASK3-eGFP fusion proteins was exposed to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability was assessed with cell counting Kit-8 (CCK-8).
RESULTSAll the results of identification by PCR, digestion with restriction endonuclease and sequencing indicated that the recombinant eukaryotic expression plasmid pEGFP-TASK3 was constructed correctly. The stable SH-SY5Y cell line expressing EGFP tagged-TASK3 fusion protein was successfully established. Exposure of the wild type SH-SY5Y cells and the stable SH-SY5Y-GFP tag-TASK3 cell line to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability of two group cells significantly reduced with pH declining, and the difference was statistically significant (P < 0.05). Compared with wild type SH-SY5Y cells, the cell viability of stable SH-SYSY-GFP tag-TASK3 cell line increased significantly with the same pH media, and the difference was statistically significant (P < 0.05).
CONCLUSIONThe eukaryotic expression vector pEGFP-TASK3 is successfully constructed and the cell line stably expressing TASK3-eGFP fusion is established which is important for their fundamental research and potential applications.
Blotting, Western ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; Polymerase Chain Reaction ; Potassium Channels, Tandem Pore Domain ; genetics ; Transfection
7.Genotypes and serotypes of avian infectious bronchitis viruses isolated during 2009-2011 in Guangxi, China.
Li-Li QIN ; Meng LI ; Rong SUN ; Zhi-Jin WU ; Kun HE ; Mei-Lan MO ; Tian-Chao WEI ; Ping WEI
Chinese Journal of Virology 2014;30(2):162-170
In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals
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Antibodies, Viral
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immunology
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Chick Embryo
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Chickens
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China
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epidemiology
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Coronavirus Infections
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epidemiology
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immunology
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veterinary
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virology
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Infectious bronchitis virus
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classification
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genetics
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immunology
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isolation & purification
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Molecular Sequence Data
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Phylogeny
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Poultry Diseases
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epidemiology
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immunology
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virology
9.A new flavone from stems of Garcinia bracteata and its anti-TMV activity.
Li-mei LI ; Jun ZHOU ; Jie LOU ; Yue-de WANG ; Kun ZHOU ; Wei DONG ; Xue-mei GAO ; Qiu-fen HU ; Zhi-yong JIANG
China Journal of Chinese Materia Medica 2015;40(21):4205-4207
A phytochemical investigation on the stems of Garcinia bracteata collected from Xishuangbanna resulted in the isolation of a new flavone. By analysis of the HRESIMS, IR, UV, 1D and 2D NMR spectra, the structure of the new compound was determined as 7-methoxy-4',6-dihydroxy-8-isobutyryl-flavone(1). Compound 1 was also tested for its anti-tobacco mosaic virus(TMV) activity. Results suggested the 1 possessed remarkable anti-TMV activity, with an inhibition rate of 28.2%.
Antiviral Agents
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chemistry
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isolation & purification
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pharmacology
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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pharmacology
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Flavones
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chemistry
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isolation & purification
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pharmacology
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Garcinia
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chemistry
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Magnetic Resonance Spectroscopy
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Plant Leaves
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chemistry
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Tobacco Mosaic Virus
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drug effects
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growth & development
10.Applicability of the multiplex quantitative antibody array system for early diagnosis of hepatocellular carcinoma.
Dong-mei GAO ; Lu SUN ; Kun GUO ; Yan LI ; Yin-kun LIU ; Xiao-nan KANG
Chinese Journal of Hepatology 2012;20(10):785-788
OBJECTIVETo develop an early and accurate detection method for hepatocellular carcinoma (HCC) based on detection of tumor-associated serum markers using a multiplex quantitative antibody array.
METHODSThe double-antibody sandwich principle was used to establish an antibody array composed of eight cancer-related serum markers, including alpha-fetoprotein (AFP), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), transforming growth factor-beta 1 (TGF-b1), and vascular endothelial growth factor (VEGF). Serum samples from 160 cases of clinically diagnosed HCC and from 58 cases of liver cirrhosis (LC; controls) were obtained to test the array. Sixty percent of the samples were randomly selected for use as the training set (HCC, n = 96; LC, n = 36), and the remaining 40% was used as the test set (HCC, n = 64; LC, n = 22). The SPSS statistical software was used to perform logistic regression analysis and to create a diagnostic model.
RESULTSWhen used with the training set, the model had sensitivity of 93.3%, specificity of 83.3%, and accuracy of 90.9%. When used with the test set, the model had sensitivity of 89.0%, specificity of 77.3%, and accuracy of 86.0%. The traditional serum AFP value (cut-off value of 20 ng/mL) showed 70.0% diagnostic sensitivity, 59.0% specificity, and 64.0% accuracy.
CONCLUSIONThe newly developed multiplex quantitative antibody detection system has high sensitivity and specificity. The diagnostic model with AFP and seven other cancer-related factors was superior to the traditional AFP only approach for early diagnosis of liver cancer, indicating its potential clinical value.
Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Early Diagnosis ; Female ; Humans ; Liver Neoplasms ; diagnosis ; Male ; Microchip Analytical Procedures ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; immunology