2.Prescription compatibility effect on the major components absorption in danshen extract and their identification.
Li LIN ; Jianxun LIU ; Ying ZHANG ; Chaoyi MAO ; Changling DUAN ; Jianhua FU ; Chengren LIN
Acta Pharmaceutica Sinica 2010;45(10):1290-5
An improved everted gut sac method was applied to the study of prescription compatibility effect on the major components in Danshen extracts. With the separation and detection by HPLC-ECD, 5 major peaks could be detected in intestinal absorbed solution after prescription administration. Following the identification by HPLC-MS/MS, peak 2, 3, 4, and 5 were rosmaric acid, lithospermic acid, salvianolic acid B, and salvianolic acid A, respectively, which also confirmed with reference standards of those components. Through paralleling substance identification, peak 2, 3, 4, and 5 could be found as the major components in Danshen extracts, except Salvianolic acid E which is undetectable in intestinal solution. The contents of peak 2, 3, and 4 did not show difference before and after compatible prescription administrated, where the peak 5 had a significant increase in the same process. Those results revealed that peak 5, salvianolic acid A, might lead to an increasing pharmacological effect after prescription compatibility.
3.Simultaneous Determination of 10 Flavonoids in Astragalus membranaceus by HPLC
Yan ZHANG ; Lin DONG ; Jingjiao YONG ; Fuying MAO ; Lei YIN ; Xueyan FU
China Pharmacy 2017;28(21):2970-2973
OBJECTIVE:To establish a method for simultaneous determination of 10 flavonoids in Astragalus membranaceus. METHODS:HPLC method was adopted. The determination was performed on Agilent SB-C18 column with mobile phase consisted of acetonitrile-0.3% formic acid (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm, and the column temperature was 35 ℃. The sample size was 10 μL. RESULTS:The linear ranges of calycosin-7-O-glucoside,iso-quercitrin,genistin,ononin,calycosin,quercetin,genistein,kaempferol,isorhamnetin and formononetion were 0.03029-1.5145μg (r=0.9994),0.01500-0.7500 μg(r=0.9995),0.00739-0.3695 μg(r=0.9991),0.12011-6.0055 μg(r=0.9998),0.03836-1.918 μg (r=0.9999),0.02989-1.4945 μg(r=0.9995),0.00704-0.352 μg(r=0.9994),0.01683-0.8415 μg(r=0.9995),0.00454-0.227μg(r=0.9999),0.01336-0.668 μg(r=0.9999),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0% . The recoveries were 99.55% -100.45%(RSD=0.36% ,n=6) ,99.34% -101.00%(RSD=0.59% ,n=6) , 98.05%-100.36%(RSD=1.27%,n=6),99.73%-100.13%(RSD=0.18%,n=6),99.70%-100.30%(RSD=0.22%,n=6), 99.67%-103.27%(RSD=1.37%,n=6),98.13%-104.41%(RSD=2.37%,n=6),96.35%-100.06%(RSD=1.46%,n=6), 99.47%-101.13%(RSD=0.60%,n=6),99.70%-100.06%(RSD=0.15%,n=6),respectively. CONCLUSIONS:This method is convenient,sensitive,stable and reproducible,can be used for simultaneous determination of 10 flavonoids in A. membranaceus.
4.Rapid detection of clinical pathogenic bacteria and antimicrobial-resistant genes using DNA chip
Hongju MAO ; Dongfang LIN ; Xiaogang XU ; Honglian ZHANG ; Yingyuan ZHANG ; Fu WANG ; Jianlong ZHAO ;
Chinese Journal of Infectious Diseases 2001;0(06):-
Objective To detect the clinical pathogenic bacteria and antimicrobial-resistant genes quickly and sensitively using DNA chip.Methods Based on the analysis of 23S rRNA gene se- quences and other genes sequences associated with antimicrobial resistance(SHV<CTX_M),oligo nucleotide microarray was designed according to different bacteria and antimicrobial-resistant genes. The DNA fragments were amplified by labeling Cy5 fluorescence and detect clinical pathogenic bacte- rias and antimicrobial-resistant genes by hybridization.Results The result of detection(10~3-10~6 bac- teria/ml)was consistent with that of some documents in domestic and overseas under ideal circum- stances of detecting bacteria genomic DNA by the Reagent Box.And it was specific and reproducible when the detection system were evaluated with some clinical isolates and drug-resistant standard strain.DNA chip could identify 16 species and 7 generics including common diverse clinical pathogenic bacteria,and could detect the drug-resistant of extended spectrum?lactamase gene simultaneously. Conclusions The methods that we have established DNA chip is a sensitive,specific and reproducable tool for supplying routine methods.
5.Clinical application of the free superficial peroneal artery perforator flap.
Xiao-Dong YANG ; Yang-Wu LIU ; Jin YANG ; Gen-Fu ZHANG ; Mao-Chao DING ; Jin MEI ; Mao-Lin TANG
Chinese Journal of Plastic Surgery 2012;28(2):88-91
OBJECTIVETo investigate the applied anatomy of the superficial peroneal artery perforator flap and report the clinical results of repairing the soft tissue defects with free perforator flaps.
METHODS15 fresh cadavers were injected with a modified lead oxide-gelatin mixture for three-dimensional visualization reconstruction using a 16-slice spiral computed tomography scanner and specialized software (Materiaise's interactive medical image control system, MIMICS). The origin, course and distribution of the superficial peroneal artery perforator in the anterolateral leg region were observed. Clinically 6 cases with hand defects and 6 cases with feet defects were treated with free superficial peroneal artery perforator flap transplantation. The defect size ranged from 3.0 cm x 4.5 cm to 5.0 cm x 11.0 cm.
RESULTSThe diameter of the superficial peroneal artery is (1.2 +/- 0.3) mm at its origin from the anterior tibial artery 5 cm below the fibula head. It is (5.6 +/- 1.8) cm in length. This artery is truly anastomosed with other perforators to form the chain of superficial peroneal nerve accessory artery. The superficial peroneal artery perforators [outer diameter (0.7 +/- 0.2) mm] with a vein are in the anterolateral leg region, supplying the skin in proximal-middle region. All the 12 cases were treated successfully. The clinical results were satisfactory after 3-12 months of following-up.
CONCLUSIONSThe superficial peroneal artery perforator flap has constantly, reliable blood supply, and good texture. It is a good option for repairing soft-tissue defect with free transfer.
Cadaver ; Fibula ; Foot ; Foot Injuries ; surgery ; Free Tissue Flaps ; blood supply ; innervation ; transplantation ; Hand Injuries ; surgery ; Humans ; Leg ; Perforator Flap ; blood supply ; innervation ; transplantation ; Peroneal Nerve ; Soft Tissue Injuries ; surgery ; Tibial Arteries
6.Effect of sleep recovery on the executive function of the brain: an ERP study
Fu-gui, WANG ; Jian-lin, QI ; Yong-cong, SHAO ; En-mao, YE ; Guo-hua, BI ; Nuo-min, LI ; Zheng, YANG
Bulletin of The Academy of Military Medical Sciences 2010;34(1):46-50
Objective To explore the effect of recovery sleep on the executive function after 36 h of total sleep deprivation by event related potential technology.Methods Thirteen healthy male college students participated in two trials. At the first trial normal sleep as control was investigated. At the second trial participants experienced 36 h of sleep deprivation and then accepted 8 h recovery sleep. In each trial six Go/Nogo tests were employed to test the executive control function and the ERP data were recorded. Results There was no statistical difference in behavior and ERP results at each time point as the subjects had normal sleep. After 36 h of sleep deprivation, the behavior results were statistically significant when compared to the baseline. The amplitude and latency of Nogo-N2, Nogo-P3 on Fz electrode, the amplitude and latency of Nogo-P3 on Cz electrode showed statistical significance when compared to the baseline. After 8 h recovery sleep, the average correct reaction time and the Go correct reaction rate had statistical significance compared to 36 h value. The amplitude of Nogo-N2 and Nogo-P3 had no statistical significance compared to the baseline.However,it was of statistical significance[(-6.80 3.95)vs(-3.37 2.63)μV,(10.63±6.62)vs(5.63±5.45)μV,(9.49±7.37)vs(6.08±6.56)μV] compared to 36 h value. The latency of the recovery value of Nogo-N2 and Nogo-P3 was statistically significant[(254.14±15.55)vs(243.08±13.97)ms(382.14±41.07)vs(349.17±30.36)ms,(369.86±26.48)vs(347.48±29.24)ms]compared to the baseline.Conclusion As the time of sleep deprivation is prolonged, the executive function is impaired and the executive function is not completely recovered after 8 h recovery sleep.
7.Effect of schisandrin B on lung mRNA expression of transforming growth factor-beta1 signal transduction molecule in rat lungs exposed to silica.
Lin-Hua FAN ; Tian-Fu LIU ; Min GUO ; Mao-Lin LIU ; Zhi-Ping WANG ; Su-Jin SI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(4):255-259
OBJECTIVETo investigate the effects of schisandrin B (Sch-B) on expression of transforming growth factor-beta1 (TGF-beta1) and signal transduction molecule mRNA in rat lungs exposed to SiO2, and explore the intervention mechanism of Sch-B on pulmonary fibrosis induced by SiO2.
METHODSNinety six Wistar rats were randomly divided into control (normal saline) group, SiO2 group and SiO2 plus Sch-B group. The rats were exposed to SiO2 by direct tracheal instillation to establish the silicotic animal models. SiO2 group and SiO2 plus Sch-B group were treated with 1 ml SiO2 (50 mg/ml) for each rat From the first day after model establishment, SiO2 plus Sch-B group were orally given Sch-B (80 mg/kg) a day, control group and silica group were orally given olive oil. On the 3rd, 7th, 14th and 28th days after treatment, 8 rats in each group were sacrificed and samples were collected. The histo-pathological examination of lung was performed by HE staining. The expression levels of TGF-beta1, TGF-betaR II and Smad4 mRNA in the lung tissues were detected by RT-PCR.
RESULTSThe results of histo-pathological examination showed that in SiO2 group, lung tissues were injured obviously; the alveolar inflammation with alveolus interval edema and inflammation cell infiltration appeared on the 3rd and 7th days; the alveolus interval became thicker, became thicker, fibroblast and collagen matrix increased markedly on 14th day; the alveolar structure was damaged, alveolar wall thickened obviously, collagen aggravation and pulmonary fibrosis displayed on 28th day. The alveolar inflammation and pulmonary fibrosis in SiO2 plus Sch-B group were significantly less than those in SiO2 group. The expressions levels of TGF-beta1 TGF-betaR II and Smad4 mRNA (TGF-1beta: 1.03 +/- 0.31, 1.33 +/- 0.39,1.08 +/- 0.26, 0.82 +/- 0.16, TGF-betaR II: 0.65 +/- 0.11, 0.80 +/- 0.16, 0.83 +/- 0.24, 0.62 +/- 0.15, Smad4:0.87 +/- 0.15, 0.68 +/- 0.11, 0.78 +/- 0.19, 0.30 +/- 0.08) in SiO2 group were significantly higher than those in the control group (TGF-beta1:0.59 +/- 0.22, 0.55 +/- 0.25, 0.56 +/- 0.20, 0.55 +/- 0.12, TGR-betaR II :0.28 +/- 0.13, 0.31 +/- 0.15, 0.34 +/- 0.15, 0.27 +/- 0.09, Smad4:0.23 +/- 0.11, 0.40 +/- 0.12, 0.39 +/- 0.12, 0.18 +/- 0.06) (P < 0.01 or P < 0.05), but the expression level of TGF-beta1 mRNA was the highest on the 7th day. The expression levels of TGF-beta1 and Smad4 mRNA (TGF-beta1:0.68 +/- 0.28, 0.88 +/- 0.25, 0.75 +/- 0.11, 0.61 +/- 0.14,Smad4:0.25 +/- 0.12, 0.45 +/- 0.09, 0.44 +/- 0.07, 0.21 +/- 0.04) in SiO2 plus Sch-B group were significantly lower than those in SiO2 group (P < 0.01 or P < 0.05 ), but there were no significant differences of the TGFbetaR II mRNA expression levels between SiO2 group and SiO2 plus Sch-B group.
CONCLUSIONSch-B can reduce the pulmonary fibrosis induced by SiO2 through inhibition of the mRNA express of TGF-beta1 and Smad4 in the lung tissue, modulating the TGF-beta1/Smad4 signal transduction pathway and inhibiting the target gene activation.
Animals ; Cyclooctanes ; pharmacology ; Female ; Lignans ; pharmacology ; Lung ; drug effects ; metabolism ; pathology ; Male ; Polycyclic Compounds ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Silicosis ; metabolism ; pathology ; Smad4 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism
8.Construction of recombinant adenovirus co-expression vector carrying the human transforming growth factor-beta1 and vascular endothelial growth factor genes and its effect on anterior cruciate ligament fibroblasts.
Xue-Lei WEI ; Lin LIN ; Yu HOU ; Xin FU ; Ji-Ying ZHANG ; Ze-Bin MAO ; Chang-Long YU
Chinese Medical Journal 2008;121(15):1426-1432
BACKGROUNDRemodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta1 (TGFbeta1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGFbeta1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.
METHODSAdenoviral vector containing TGFbeta1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGFbeta1, the expression of VEGF165 and TGFbeta1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGFbeta1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.
RESULTSThe results showed the successful construction of a recombinant co-expression adenovirus vector containing TGFbeta1 and VEGF165 genes. Co-expression of TGFbeta1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.
CONCLUSIONCo-expression of TGFbeta1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.
Adenoviridae ; genetics ; Animals ; Anterior Cruciate Ligament ; cytology ; metabolism ; Cell Movement ; Cells, Cultured ; Collagen ; genetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibroblasts ; physiology ; Fibronectins ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing
9.An analysis of hyperinsulinemia in Bartter syndrome
Shi CHEN ; Zhengpei ZENG ; Anli TONG ; Lin LU ; Ailing SONG ; Wei LIANG ; Yong FU ; Weibo XIA ; Yan JIANG ; Jiangfeng MAO ; Huabing ZHANG ; Wei LI
Chinese Journal of Internal Medicine 2011;50(2):128-131
Objective To analys hyperinsulinemia in Bartter syndrome. Methods Twenty-three cases of Bartter syndrome [age (27 ±9) years;fasting serum potassium(2. 8 ±0. 5)mmol/L], 20 patients of aldosterone-producing adenoma [APA, age (45 ± 11 ) years, fasting serum potassium ( 3.0 ± 0. 4 ) mmol/L], 20 patients of idiopathic hyperaldosteronism [IHA, age (51 ± 11 ) years, fasting serum potassium (3.4 ±0. 2)mmol/L] were diagnosed in Peking Union Medical College Hospital from September 2003 to May 2008. All patients underwent 3-hours oral glucose tolerance test(3hOGTT), postural stimulation test and calculated HOMA-insulin resistance ( HOMA-IR ) and HOMA-insulin sensitivity ( HOMA-IS ) by Homeostasis model.Results The insulin area under curve-(229.0±162.4)mIU·L-1·h] was singnificantly higher than APA group [(227.7±158.6)mIU·-1·h].But HOMA-IR in Bartter group were similar to APA group( 1.96 ± 1.14 vs 1.41 ± 0. 91 ), and HOMA-IR in APA group was lower than IHA group ( 1.96 ± 1.14 vs 2.40 ± 1.60, P < 0. 05 ). There was no deference in HOMA-IS among three groups,but APA group had lower level. In all three groups, the peak of insulin secretion was delayed. Conclusion Bartter syndrome patients commonly present with hyperinsulinemia.
10.Change of extracellular ascorbic acid in the brain cortex following ice water vestibular stimulation: an on-line electrochemical detection coupled with in vivo microdialysis sampling.
Na ZHANG ; Jun-xiu LIU ; Fu-rong MA ; Li-sheng YU ; Yu-qing LIN ; Kun LIU ; Lan-qun MAO
Chinese Medical Journal 2008;121(12):1120-1125
BACKGROUNDAscorbic acid (AA) represents one of the most important enzyme co-factors, antioxidants and neuromodulators and plays an important role in the cerebral system. Increasing evidence has suggested that AA could treat certain kinds of vertigo diseases such as Meniere's disease. To elucidate the neurochemical functions associated with AA in vertigo, the change of extracellular AA in the brain cortex following caloric vestibular stimulation (CVS) was evaluated.
METHODSAn on-line electrochemical detection was coupled with in vivo microdialysis to continuously monitor the change of extracellular AA in the primary somatosensory (SI) area of guinea pigs following a caloric vestibular stimulation. Sixteen guinea pigs were divided into three groups, i.e., experimental group with irrigation of the ear canal with ice water (0 degrees C) (n = 8), and two control groups, one with irrigation of the ear canal with warm water (38 degrees C) (n = 4) and the other with irrigation of the auricle with ice water (n = 4).
RESULTSIn the experimental group, the ice water irrigation of the left external ear canal induced a horizontal nystagmus towards the right side lasting about 45 seconds. No nystagmus was induced by warm water irrigation of the external ear canal or by ice water irrigation of the auricle. The extracellular AA concentration significantly increased following the ice water vestibular stimulation, reaching a maximum of (130 +/- 20)% (n = 8) of the basal dialysate level (2.61 +/- 0.92) micromol/L (n = 8), lasting at least for an hour. AA level did not change distinctly after the irrigation of the left external ear canal with warm water or the irrigation of the auricle with ice water.
CONCLUSIONSThe concentration of extracellular AA in the brain cortex of the SI area increased following the ice water vestibular stimulation. This demonstration may be useful for the investigation of the neurochemical processes associated with AA in the process of vertigo.
Animals ; Ascorbic Acid ; analysis ; Cerebral Cortex ; metabolism ; Electrochemistry ; methods ; Extracellular Space ; metabolism ; Guinea Pigs ; Ice ; Male ; Microdialysis ; methods ; Physical Stimulation ; methods ; Vestibule, Labyrinth ; physiopathology