1.Construction of an immortalized rat astrocyte strain with inducible enkephalin expression
Ying XU ; Yu-Ke TIAN ; Xue-Bi TIAN ;
Chinese Journal of Anesthesiology 1994;0(06):-
Objective To construct an immortalized rat astrocyte strain expressing enkephalin which can be regulated by doxycycline.Methods pRevTet-On and recombinant plasmid pRevTRE/hPPE were transfected into packaging cell PT67 respectively by standard lipofectamine methods.The supernatant containing RevTet-On virus was used to infect immortalized rat astrocyte strain(IAST)and RevTRE/hPPE virus was used to infect the IAST/Tet-On cell.Immortalized rat astrocyte strain that stably expressed Tet-regulated enkephalin was established. hPPE gene expression level of IAST/Tet-On/hPPE cell strain was detected by real time-PCR,immuno- cytochemistry and radio-immunoassay.Results Real time-PCR,immuno-cytochemistry and radio-immunoassay confirmed the level of enkephalin expression was regulated by doxycycline in a dose-dependent manner.Conclusion An immortalized rat astrocyte strain which secrets enkephalin as controlled by doxycycline has been constructed successfully.
2.Effect of Qingguang'an on elastic fiber, MMP- 7, TlMP - 1 in scarring area of filtration canal after glaucoma surgery
Yuan-Bi, LI ; Qing-Hua, PENG ; Xue-Si, HUANG ; Xiao-Liu, CHEN ; Han-Yu, TAN
International Eye Science 2015;(1):20-25
?AlM: To observe the effect of Qingguang'an on elastic fiber, MMP-7, TlMP-1 in scarring area of filtration canal after glaucoma surgery through the four Qingguang'an effective groups and Qingguang'an granules, to discuss and compare their mechanism of action on scarring area of filtration canal.?METHODS:Four effective components of Qingguang'an were used in groups D, E, F, G and H after glaucoma surgery, compared with group A ( blank ) , group B (model) and group C ( MMC) to observe the effect of elastic fiber, MMP-7, TlMP-1 in scarring filtration canal.?RESULTS:Compared with the preoperative basic lOP and 2d , 1, 2, 4wk postoperative lOP of groups C, E and H, the lOP of three group rose up slower than other groups, and kept the lowest data at 28d. There was significant difference compared with the rest of A, B, D, F, G groups (P<0. 05). The area and density of elastic fiber in surgery group were significantly different with that of black control group ( P<0. 05 ), but there were no statistical differences between groups C and H, groups C and F, groups H and E (P>0. 05). The difference was statistically significant among other groups (P<0. 01).?CONCLUSlON:The scarring area of filtration canal after glaucoma surgery is the major reason which lead to the failure of surgery. Qingguang'an effective group 2, Qingguang'an granules and MMC could reduced the scar tissue by restrained the elastic fiber, TlMP - 1 and increased the MMP-7. By observing the experimental results that both Qingguang'an effective group 2 and Qingguang'an granules could restrained the scarring area of filtration canal, the effects were unbiased, Qingguang'an granules group is better than effective group 2.
3.Analysis of the glycosylation heterogeneity of recombinant human pro-urokinase using UPLC-MS
Lei TAO ; Lei YU ; You-xue DING ; Hua BI ; Chun-ming RAO
Acta Pharmaceutica Sinica 2020;55(11):2713-2718
The glycosylation heterogeneity of recombinant human pro-urokinase (pro-UK) was assessed using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Firstly, the source of heterogeneity was determined by measuring the
4.Dynamic changes of blood flow in frontal cortex of diabetic rats
yan Ming YANG ; Yue WANG ; tian Tian BI ; ying Yu BI ; xue Dong WANG ; Ying ZHAO
Chinese Pharmacological Bulletin 2017;33(11):1517-1523
Aim To investigate the changes of cerebral blood flow in rats with diabetic cognitive impairment by two-channel laser Doppler flowmeter,and to explore the changes of cerebral blood flow in diabetic rats with cognitive impairment and to investigate the changes of cerebral blood flow lesions and the central nervous system function changes in the study to provide pre-foundation.Methods Thirty-four male Wistar rats were randomly divided into two groups:blank control group and diabetic model group.The rats in the model group were treated with streptozotocin (STZ) 60 mg · kg-1,and the glucose oxidase method was used to determine fasting blood glucose ≥ 16.7 mmol · L-1 as the standard of the model.The Water maze was used to observe the behavioral changes of diabetic rats.Dual-channel laser doppler flowmeter was used to measure cerebral blood flow in diabetic rats with cognitive impairment.Another 34 male Wistar rats were randomly divided into two groups:control group and diabetic model group.Dual-channel laser doppler flowmeter was used to dynamically monitor cerebral blood flow on 0 d,7 d,14 d,21 d,28 d,35 d,42 d,56 d and 75 d.ELISA was applied to detect the concentration of iNOS,eNOS and ET-1 in cerebrospinal fluid.Results Morris water maze test showed that the time of the platform (latency) was significantly longer than that of the blank control group(P <0.05).The cerebral blood flow/100 g of diabetic rats with cognitive impairment was significantly lower than that of the control group (P < O.05),and the blood flow in the model group was significantly lower than that in the model group (P < 0.05).Compared with control group,iNOS and eNOS concentrations were markedly elevated,while ET-1 concentration obviously decreased.Conclusions The decrease of blood flow in the frontal cortex of diabetic rats with cognitive impairment suggests that it may be one of the factors leading to cognitive impairment in diabetes mellitus.Cerebral blood flow reduction occurs in the early stages of diabetes,followed by no significant deterioration.Cerebral blood flow may not be related to the changes of NO and ET-1,but the trend of cerebral blood flow may be related to the change of the two.
5.Rapid identification of six chemical constituents in Guizhi Fuling capsule by DART-Q-TOF-MS.
Zhang WEI ; Wang XUE ; Yan-jing-ping ; Yan-jing LI ; Yu-an BI ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(21):4118-4122
In order to establish a rapid method for identifying six constituents in Guizhi Fuling capsule, Q-TOF with DART ion source was used to perform the direct analysis of compounds in Guizhi Fuling capsule. The DART sampler delivery rate was 0.2 mm s(-1). The temperature of helium gas of DART was 450 degrees C. The capillary voltage was kept at 1 000 V. The temperature of the drying gas of Agilent 6538 Q-TOF MS was set at 350 degrees C. The flow rate of the drying gas of MS was set at 3.5 L x min(-1). The MS scan range was m/z 50-1 000. Based on accurate mass measurements and the elemental compositions of the product ions and fragmentation patterns of reference conpounds, six components, amygdalin, paeonol, paeoniflorin, cinnamic acids, gallic acid, benzoic acid were identified rapidly. The method can rapidly identify six chemical constituents in three batch of Guizhi Fuling capsule. The DART-Q-TOF-MS method is simple, rapid and specific and it can be used for rapid identification and characterization of compounds in traditional Chinese medicines.
Capsules
;
Drugs, Chinese Herbal
;
analysis
;
Mass Spectrometry
;
methods
6.Determination of eleven major components and fingerprint chromatography for Reduning injection by UPLC.
Sha WU ; Xue WANG ; Ya-nan WU ; Qi-an LIU ; Jian-xiong WU ; Yu-an BI ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(24):4804-4810
A reliable method for simultaneous determinition of eleven representative components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin) in combination of chromatographic fingerpint analysis for Reduning injection was developed by ultra high-performance liquid chromatography (UPLC). The method was performed on an Agilent ZORBAX SB-C18 anlytical column (3. 0 mm x 100 mm, 1. 8 µm) with a guard column of Agilent UPLC Guard ZORBAX SB-C18 (3.0 mm x 5 mm) at the column temperature of 30 °C. The gradient mobile phase consisted of acetonitrile (A)-0. 1% phosphoric acid (B) with a flow rate of 0. 4 mL . min-1. The injection volumn was 2 µL. The detection wavelengths were set at 324 nm and 238 nm for quantit tive analysis and 225 nm for fingerpint chromatography. Neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin were baseline seperated with good linearity relationships (r >0. 999) between concentration and peak areas over the linear ranges. The average recoverys of the investigated compounds were 103.5%, 100. 2%, 103. 3%, 102. 8%, 101. 3%, 102. 8%, 97. 36%, 99. 62%, 98. 16%, 102. 8%, 99. 27%, respectively. Reduning injection of forty-five batches was analyzed by UPLC finge print chromatography. Thirty batches were selected to generate the reference fringerprint chromatography with fourteen common peaks. The similarity values between the reference fringerprint chromatography and the remaining fifteen batches were higher than 0. 99. The developed method was fast, accurate and sensitive. It could be used as a reference for the quality control of multiple components determination and fingerprint chromatography for Reduning injection in future.
Chlorogenic Acid
;
chemistry
;
isolation & purification
;
standards
;
Chromatography, High Pressure Liquid
;
methods
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
standards
;
Glucosides
;
chemistry
;
isolation & purification
;
standards
;
Iridoids
;
chemistry
;
isolation & purification
;
standards
;
Quality Control
;
Reference Standards
;
Reproducibility of Results
;
Sensitivity and Specificity
;
Time Factors
7.Construction and identification of immortalized rat astrocyte cell line expressing enkephalin.
Ying XU ; Yu-ke TIAN ; Xue-bi TIAN ; Ke AN ; Hui YANG
Chinese Journal of Traumatology 2007;10(6):353-356
OBJECTIVETo provide a sound cell source for further ex-vivo gene therapy for chronic pain, we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline.
METHODSRetrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline. The hPPE gene expression level of immoralized astroyte cells (IAC)/ hPPE was detected by RT-PCR, indirect immunofluorescence staining and radioimmunoassay.
RESULTSIAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline.
CONCLUSIONAn immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully, which provides a research basis for transgenic cell transplantation for analgesia.
Animals ; Anti-Bacterial Agents ; pharmacology ; Astrocytes ; Cell Line, Transformed ; Chronic Disease ; Doxycycline ; pharmacology ; Enkephalins ; genetics ; metabolism ; Genetic Therapy ; methods ; Genetic Vectors ; Neurotransmitter Agents ; genetics ; metabolism ; Pain Management ; Protein Precursors ; genetics ; metabolism ; Rats ; Retroviridae ; genetics
8.The introduction of breastfeeding in children under age of three in the counties of western China in 2005.
Yi-jun KANG ; Hong YAN ; Quan-li WANG ; Qiang LI ; Sheng-bin XIAO ; Yu-xue BI ; Hong XIE
Chinese Journal of Epidemiology 2007;28(2):109-114
OBJECTIVEThe study was to describe the breastfeeding status of children under the age of three in counties of western China and to provide evidence to the government for decision-making on intervention.
METHODSA cross-sectional study with probability-proportional-to-size (PPS) sampling method was used. The information on breastfeeding was obtained through memory of the mothers. Fourteen thousand and seventy-seven children were studied. Data on breastfed status in counties of western China was compared with those of the children from the survey of the counties of western China in 2001.
RESULTSThe breastfeeding rate of children under 3 years old in western China was 96.5%. However, the overall breastfed rate of children under 6 months were only 33.4%, with rates of 11.4% and 22.0% on exclusively and predominantly breastfed groups respectively. Timely first-suckling rate was 43.5% with the continued breastfeeding rate (1 year) as 64.9%, but the continued breastfeeding rate (2 year) was only 9.7%. Reasons causing mothers to wean would include according to her own intention and to be able to attend the physical labor while exclusive breastfeeding under 6 months was for the growth and development of children, which might reduce the two-week prevalence of diarrhea. Major risks of exclusive breastfeeding of children under 6 months were seen as: level of education of the mothers, economic depression of the counties and mother's nationality (if as minority).
CONCLUSIONMost of the children were ever or being breastfed at the time of interview with timely first-suckling took place earlier than in 2001. However continued breastfeeding did not last long. During these five years, the exclusive breastfeeding rate had been at low level, especially at the economic depression and the minority area.
Breast Feeding ; epidemiology ; Child, Preschool ; China ; epidemiology ; ethnology ; Humans ; Infant ; Infant, Newborn ; Weaning
9.The treatment of lower eyelid retraction at the different degree.
Ling-Li GUO ; Hong-Da BI ; Chun-Yu XUE ; Jun-Hui LI ; Chao YANG ; Xin XING
Chinese Journal of Plastic Surgery 2010;26(3):168-171
OBJECTIVETo explore the effective methods for the correction of lower eyelid retraction at different degree.
METHODS258 patients with lower eyelid retraction were treated in our department since 1999. The lower eyelid retraction could be divided into mild, moderate and severe degree. The lateral canthal anchoring (n = 150), Hamra's lower eyelid blepharoplasty (n = 80) and translid cheek lifting (n = 28) were adopted according to the severity. The therapeutic effect for different degree of lower eyelid retraction was compared.
RESULTS98 patients were followed up for 3-12 months, including 51 patients of mild degree, 29 patients of moderate degree, and 18 patients of severe degree. The retraction were corrected completely in 91 patients. The lower eyelid was repositioned to the level of inferior limbus without inferior scleral show when eyes opened. The palpebral fissure could close completely. The blunt lateral canthus turned to be acute and the scar was inconspicious. The retraction was improved, but not corrected completely in 3 patients of moderate degree and 4 patients of severe degree. The complications included petechiae, chemosis, and so on.
CONCLUSIONSThe lower eyelid retraction can be corrected effectively if the appropriate techniques are performed according to the degree of retraction.
Adult ; Aged ; Blepharoplasty ; methods ; Eyelid Diseases ; surgery ; Eyelids ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Treatment Outcome
10.Effect of ischemia/hypoxia on mesenteric vasomotor function in spontaneously hypertensive rats and its possible mechanism.
Ming ZHAO ; Xiao-Jiang YU ; Hong-Li ZHANG ; Xue-Yuan BI ; Hao HU ; Wei-Jin ZANG
Acta Physiologica Sinica 2011;63(6):540-548
Hypertension is a common cardiovascular disease and can induce many complications, such as stroke and coronary heart disease. The purpose of the present study was to investigate the effect of ischemia/hypoxia on mesenteric artery vasomotor function in spontaneously hypertensive rats (SHR). Rat mesenteric arterial rings were cultured in modified ischemia-mimetic solution in a hypoxia incubator for a certain time period. Isometric tension changes of isolated mesenteric arterial rings were recorded continuously by a myograph system. The results obtained were as follows: In SHR group, the maximum contractions to KCl and phenylephrine (PE) were increased, and the maximum relaxation to acetylcholine (ACh) was decreased, compared to those in Wistar-Kyoto (WKY) rats group. Compared with SHR group and WKY with acute ischemia/hypoxia (WKY+H) group, SHR with acute ischemia/hypoxia (SHR+H) increased the maximum contractions induced by KCl and PE and inhibited the maximum relaxations by ACh. In SHR+H and SHR groups, the vasodilation induced by ACh was unaffected by N(G)-nitro-L-arginine methylester (L-NAME), whereas in WKY group, the relaxation to ACh was attenuated by L-NAME. CaCl2-induced contraction in depolarized rings in SHR+H group significantly shifted to the left compared with SHR group. In Ca(2+)-free K-H solution, the maximum contractions induced by PE and caffeine were increased in SHR+H group compared to those in WKY+H group; the PE- and caffeine-induced contractions were also enhanced in SHR group versus WKY group; the maximum contraction induced by PE was significantly increased in SHR+H group versus SHR group. These findings suggest that acute ischemia/hypoxia aggravates mesenteric artery dysfunction in SHR. The mechanism may be related to the decreased NO generation and increased sarcoplasmic reticulum Ca(2+) release.
Animals
;
Calcium
;
metabolism
;
Endothelium, Vascular
;
metabolism
;
physiopathology
;
Hypertension
;
physiopathology
;
Hypoxia
;
physiopathology
;
In Vitro Techniques
;
Male
;
Mesenteric Arteries
;
physiopathology
;
Muscle, Smooth, Vascular
;
physiopathology
;
Nitric Oxide
;
biosynthesis
;
Rats
;
Rats, Inbred SHR
;
Rats, Inbred WKY
;
Vasomotor System
;
physiopathology