1.Alteration of biological and immunological properties of aortic homografts after de-endothelialization and decellularization
Chinese Journal of Tissue Engineering Research 2014;(33):5249-5256
BACKGROUND:Aortic root homograft has been widely used, but it stil has a high incidence of calcification and degeneration after operation.
OBJECTIVE:To investigate the alteration of biological and immunological properties of rabbit aortic root homograft treated with de-endothelialization and decellularization methods, and to find a better decellularization method.
METHODS:Aortic root homografts obtained from rabbits were randomly divided into fresh group, de-endothelialization group, and decellularization group. The morphology of homografts was evaluated by hematoxylin-eosin, Masson and VG stainings. The biological properties such as thickness, tensile strength, broken extension rate, and heat shrinkage temperature were detected. The immunological features (inflammatory cellinfiltration, calcium content and CD152 expression) were determined with immunohistochemistry and flame atomic absorption spectrometry.
RESULTS AND CONCLUSION:As demonstrated in morphological observation, protocols of de-endothelialization and decellularization methods we used attained our desired aim of decellularization perfectly. Tensile strength and broken extension rate of decellularization group were decreased significantly compared with fresh group (P<0.01), while the difference between de-endothelialization group and fresh group was not significant (P>0.05). Hematoxylin-eosin staining showed that, inflammatory reaction was the most apparent in fresh group, showing a large amount of inflammatory cells infiltration;compared with fresh group, inflammatory reaction was significantly attenuated in de-endothelialization group, the number of inflammatory cells was significantly decreased. The inflammatory reaction was the lowest in the decellularization group, only inflammatory cells infiltrated the tissue edge. Immunohistochemistry showed that, CD152 expression was significantly increased in de-endothelialization and decellularization groups compared with fresh group (P<0.05). Calcium content in de-endothelialization and decellularization groups was significantly decreased compared with fresh group (P<0.05). It could be concluded that the immunogenicity of de-endothelialized and decellularized aortic homografts is decreased significantly, and the de-endothelialization method could better maintain the tensile property.
2.Determination of 5 aphrodisiacs illegally adulterated into Chinese proprietary medicine by LC/MS(n) Trap
Chang LIU ; Jing XIA ; Qing HU ; Shen JI
Chinese Traditional Patent Medicine 1992;0(09):-
AIM: To determine 5 different aphrodisiacs,including apomorphine,yohimbine,vardenafil,sildenafil and alprostadil,adulterated into Chinese proprietary medicine by LC/MS(n),and build a laboratory library including MS(n) spectra and retention time. METHODS: ZORBAX SB C_18 column(150 mm?4.6 mm,5 ?m)was applied.The mobile phase consisted of acetonitrile and 0.5% formic acid changing by linear gradient elution,and the flow rate was 1.0 mL/min.The ion trap mass spectrometer with positive ion mode electrospray(ESI?) was applied. RESULTS: In the mass spectrum apomorphine,yohimbine,vardenafil and sildenafil showed abundant pseudomolecular ion +,and alprostadil showed abundant pseudomolecular ion -.All the MS(n) spectra showed characteristic product ion. CONCLUSION: It is proved that the method is simple,reliable and suitable to determin aphrodisiac adulterated to health food.
3.Assessment of regional left ventricular myocardial function in rats after acute occlusion of left anterior descending artery by two-dimensional speckle tracking imaging.
Qian, FU ; Mingxing, XIE ; Jing, WANG ; Xinfang, WANG ; Qing, LV ; Xiaofang, LU ; Lingyun, FANG ; Long, CHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(6):786-90
This study evaluated the change in regional left ventricular myocardial function in rats following acute occlusion of the left anterior descending coronary artery (LAD) by using two-dimensional speckle tracking imaging (2D-STI). Sixty Wistar rats were randomly divided into two groups, a myocardial infarction (MI) group, in which 50 rats were subjected to LAD occlusion for 30-45 min, and a sham-operated (SHAM) group that contained 10 rats serving as control. Echocardiography was performed at baseline and 1, 4 and 8 week(s) after the operation. High frequency two-dimensional images of left ventricular short axis at papillary muscle level were recorded. Peak systolic radial strain (PRS) and circumferential strain (PCS) were measured in the mid-ventricle in short-axis view by using EchoPAC workstation. Left ventricular internal diameter at diastole (LVIDd) and systole (LVIDs), fractional shortening (FS), ejection fraction (EF) and left ventricular mass (LVM) were measured by anatomical M-model echocardiography. Infarct size was measured using triphenyl tetrazolium chloride (TTC) staining 1 week and 8 weeks after the operation. Fibrosis of left ventricular myocardium was displayed using Van Gieson staining 1 week after the infarction. In terms of the TTC staining results, the left ventricle fell into three categories: infarcted, peri-infarcted and remote myocardial regions. Compared with those at baseline and in the SHAM group, (1) PRS and PCS in the infarcted, peri-infarcted and remote myocardial regions were significantly decreased in the MI group within 1 week after the operation (P<0.05) and the low levels lasted 8 weeks; (2) Compared with those at baseline, LVIDd, LVIDs, FS, EF and LVM in the MI group showed no significant difference 1 week after the operation (P>0.05). However, LVIDd, LVIDs and LVM were increased significantly 4 and 8 weeks after the operation (P<0.05), and FS and EF were decreased substantially (P<0.05). Van Gieson staining showed that fibrosis developed in all the three myocardial regions to varying degrees. It is concluded that 2D-STI is non-invasive and can be used to assess regional function of myocardium with different blood supply in rats following acute occlusion of the LAD, and can be used as a sensitive and reliable means to follow up the process of left ventricular remodeling.
4.Inhibitory control dysfunction of late-onset depression: an event-related fMRI study
Junliang YUAN ; Jing XU ; Bingwei ZHANG ; Jianlin WU ; Qing ZHANG ; Lifei MA ; Yi CHANG
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(10):891-893
Objective To investigate the neural circuit of inhibitory control in late-onset depressed patients(LOD) by functional magnetic resonance imaging(fMRI). Methods Fourteen late-onset depressed patients (LOD group) and thirteen elderly healthy subjects( control group) were recruited. The two groups were age, gender, and education matched. All the subjects performed a visual Go/Nogo task during the fMRI scan. Erect or inverted isosceles triangular figures were used for stimuli. The two groups were instructed to press a button as quickly and correctly as possible when the erect triangular figures(Go) were presented, but not to response when the inverted triangular figures(Nogo) were presented. The differences of brain activation between the two groups were compared. Results ( 1 ) During Go trials, there were no significant differences in reaction time and hit rate between the two groups (P > 0.05 ). During Nogo trials, however, the late-onset depressed patients showed much higher false alarm rate(0.09 ±0.06) compared with control group(0.04 ±0.02) (P<0.05=. (2) During Go trials , LOD group showed significantly greater activity in left postcentral gyrus, left inferior parietal lobule, right precentral gyrus, left paracentral lobule, right inferior parietal lobule, right anterior cingulate cortex, left middle frontal gyrus, right middle frontal gyrus, right superior frontal gyrus compared with the control group. Whereas during Nogo trials, LOD group exhibited greater activity in left inferior parietal lobule and left middle frontal gyrus compared with the control group. Conclusion This study suggests that inhibitory control dysfunction in late-onset depressed patients may be closely related to frontostriatal circuit impairment. Over activation in left middle frontal gyrus, right middle frontal gyrus and right anterior cingulate cortex may contribute to the pathogenesis of late-onset depression.
5.Clinical outcome of arthroscopic excision of the os subfibulare in ankle pain.
Chang LIU ; Hai-sen ZHANG ; Bao-jing PEI ; Huai-liang WANG ; Hang SU ; Qing-hai WANG
China Journal of Orthopaedics and Traumatology 2016;29(2):146-148
OBJECTIVETo evaluate the clinical effect of arthroscopic excision of the os subfibulare in anterior-lateral ankle pain.
METHODSFrom December 2005 to Augest 2014, 16 patients suffering from pain associated with an os subfibulare in the anterior-lateral side of their ankles were reviewed. Among the patients,11 patients were male and 5 were female, with a mean age of (33.5 ± 15.6) years old. The mean maximum diameter of os subfibulare was (0.70 ± 0.26) cm. All the patients underwent excision of the osseous fragments, and had anatomic reconstruction of the anterior talofibular ligament if the anterior-lateral ankle was instable. The average follow-up period was (18.0 ± 4.5) months. To analyze the surgical outcome, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot pain and function scales,visual analogue scale (VAS) and Tegner activity scale were assessed preoperatively and postoperatively.
RESULTSAOFAS scales were preoperative 60.15 ± 14.52 and postoperative 92.35 ± 5.73. There was a significant difference between them (t = -8.251, P = 0.000). The mean VAS score were preoperative 7.35 ± 0.46 and postoperative 2.45 ± 0.98. Statistical significance was also notable (t = 18.105, P = 0.000). Tegner score was significantly increased from preoperative 2.87 ± 1.12 to postoperative 5.78 ± 1.06 (t= -7.548, P = 0.000).
CONCLUSIONIrrespective of the size of os subfibulare, in patients with pain or instability associated with the os subfibulare, arthroscopic excision combined with reconstruction of ther anterior talofibular ligament or not was effective in restoring ankle function and eliminating pain.
Adult ; Ankle Injuries ; surgery ; Ankle Joint ; surgery ; Arthroscopy ; methods ; Female ; Fibula ; surgery ; Humans ; Lateral Ligament, Ankle ; surgery ; Male ; Middle Aged
6.Detective Significance of Cerebral Electrical Admittance Plethysmogram in Newborns with Asphyxia
ke-ying, ZHOU ; jin-zhi, SONG ; jing-zhi, LI ; hui, LUO ; xue-chang, XIAO ; zhi-guang, LI ; ben-qing, WU
Journal of Applied Clinical Pediatrics 2006;0(20):-
Objective To study hemodynamics of cerebral blood flow in newborns with asphyxia.Method Bilateral cerebral electrical admittance plethysmogram(BCEAP) was used to explore characteristics of cerebral blood flow in 20 healthy newborns and 20 newborns with asphyxia respectively from first to fourth day after birth.Results Both the ratio of Hs to b-S(Hs/ b-S) and index of admittance differential loop(ADL) Ⅰ+Ⅱ decreased significantly in newborns with asphyxia compared to normal control from first to fourth day after birth(P
7.Effects of icariin on inflammation model stimulated by lipopolysaccharide in vitro and in vivo.
Jin-Feng WU ; Jing-Cheng DONG ; Chang-Qing XU
Chinese Journal of Integrated Traditional and Western Medicine 2009;29(4):330-334
OBJECTIVETo evaluate the anti-inflammatory effects of icariin, from aspects of pro-inflammatory cytokines, inflammatory mediators and adhesion molecules.
METHODSMouse inflammation model in vitro was established by stimulating macrophage cell line RAW264. 7 with lipopolysaccharide (LPS); and the inflammation model in vivo was established by stimulating C57BL/6J mouse with LPS. Taking dexamethasone as the positive control, both models were treated with icariin, and the cell viability in model mice was detected with CCK-8 kit; tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) in cell culture medium and serum were detected by ELISA; nitric oxide (NO) in cell culture medium by Griess Reagent method; CD11b expression on the surface of neutrophil in mice by flow cytometry, and pulmonary inflammatory cell infiltration in mice by pathological section as well.
RESULTSin vitro studies showed that icariin at the doses of 1 microg/mL, 10 microg/mL and 100 microg/mL, all displayed no cytotoxicity (P < 0.01); 10 microg/mL and 100 microg/mL icariin effectively lowered the levels of TNF-alpha and IL-6 (P < 0.01) in medium; and 100 microg/mL icariin significantly reduced level of NO (P < 0.01) in medium. in vivo studies showed that icariin at the dose of 20 mg/kg significantly lowered serum TNF-alpha and IL-6 levels (P < 0.01), reduced the average fluorescence intensity of adhesion molecules CD11b (P < 0.01), and alleviated pulmonary inflammatory cell infiltration.
CONCLUSIONIcariin is a safe and effective natural anti-inflammatory drug, its partial mechanism is possible the multiple links intervention on pro-inflammatory cytokines (TNF-alpha, IL-6), inflammatory mediators (NO) and adhesion molecules (CD11b).
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; CD11b Antigen ; metabolism ; Cell Line ; Dose-Response Relationship, Drug ; Female ; Flavonoids ; pharmacology ; Inflammation ; chemically induced ; metabolism ; Interleukin-6 ; metabolism ; Lipopolysaccharides ; Macrophages ; drug effects ; pathology ; Mice ; Mice, Inbred C57BL ; Nitric Oxide ; metabolism ; Random Allocation ; Tumor Necrosis Factor-alpha ; metabolism
8.Outcomes of surgeries for acute cervical spinal cord injury without cervical spine fracture or dislocation in young and middle-aged patients.
Qing CHANG ; Zhan-sheng DENG ; Jing CHEN
Journal of Southern Medical University 2011;31(5):919-920
OBJECTIVETo study the outcomes of surgeries for acute central cervical spinal cord injury without cervical spine fracture or dislocation in young and middle-aged patients.
METHODSThe clinical data of 58 young and middle-aged patients with acute central cervical spinal cord injury treated in our hospital between August 2005 and August 2009 were analyzed retrospectively. Of these patients, 33 (24 males and 9 females) received surgical treatment and 25 (17 males and 8 females) had conservative therapy. The ASIA grade and ASIA motor and sensory score were used for evaluation at admission and at 14 days and 1 year after the treatment. The neurological symptoms and treatment outcomes in the two groups were evaluated.
RESULTSThe proportion of patients with ASIA grade D-E and the ASIA motor and sensory scores were all significantly higher in the surgical group than in the non-surgical treatment group (P<0.05).
CONCLUSIONFor young and middle-aged patients with central cervical spinal cord injury, immediate surgery can relieve the pressure on the injured spinal cord and improve the micro-circulation to promote functional recovery of the spinal cord.
Adult ; Cervical Vertebrae ; injuries ; surgery ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Neck Injuries ; surgery ; Retrospective Studies ; Spinal Cord Injuries ; surgery ; Spinal Fractures ; surgery ; Young Adult
9.Screening and identification of indoleacetic acid producing endophytic bacterium in Panax ginseng.
Yun JIANG ; Lei TIAN ; Chang-qing CHEN ; Guan-jun ZHANG ; Tong LI ; Jing-xiu CHEN ; Xue WANG
China Journal of Chinese Materia Medica 2015;40(2):213-217
Endophytic bacteria which was producing indoleacetic acid was screened from Panax ginseng by using the Salkowski method. The active strain was also tested for its ability of nitrogen fixation by using the Ashby agar plates, the PKV plates and quantitative analysis of Mo-Sb-Ascrobiology acid colorimetry was used to measure its ability of phosphate solubilization, for its ability of potassium solubilization the silicate medium and flame spectrophotometry was used, for its ability of producing siderophores the method detecting CAS was used, for its ability of producing ACC deaminase the Alpha ketone butyric acid method was applied. And the effect on promoting growth of seed by active strain was tested. The results showed that the indoleacetic acid producing strain of JJ5-2 was obtained from 118 endophytes, which the content of indoleacetic acid was 10.2 mg x L(-1). The JJ5-2 strain also had characteristics of phosphate and potassium solubilization, nitrogen fixation, producing siderophores traits, and the promoting germination of ginseng seeds. The JJ5-2 strain was identified as Bacillus thuringiensis by analyzing morphology, physiological and biochemical properties and 16S rRNA gene sequences.
Bacteria
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isolation & purification
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metabolism
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Endophytes
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isolation & purification
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metabolism
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Indoleacetic Acids
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metabolism
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Panax
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microbiology
10.Effects of RNA interference and nolatrexed on thymidylate synthase expression and cell proliferation of human colorectal carcinoma LOVO cells.
Shu-bo TIAN ; Chang-qing JING ; Le-ping LI
Chinese Journal of Gastrointestinal Surgery 2012;15(11):1187-1191
OBJECTIVETo investigate the effect of the RNAi and the chemotherapy drugs nolatrxed on the expression of thymidylate synthase(TS) and the growth of the colorectal carcinoma LOVO cells.
METHODSThe siRNA was constructed targeting the human TS gene, and then transfected into the human colorectal cancer LOVO cells. RT-PCR and Western blot technique were used to observe the TS gene and protein expression levels, and MTT was used to detect cell proliferation after silencing the TS gene. In addition, siRNA and nolatrxed were applied to the LOVO cells to observe the TS protein expression and cell growth.
RESULTSTS siRNA significantly reduced the expression of TS gene and protein in LOVO cells, and inhibited cell growth. The IC50 value of LOVO cells was (1.46±0.25) μmol/L in TS siRNA combined with nolatrexed group, (6.81±0.31) μmol/L in the negative control group, and (6.47±0.43) μmol/L in the single nolatrexed group. After treatment of TS siRNA combined with nolatrexed on LOVO cells for 36 hours, the apoptosis index was higher than that in single TS siRNA and nolatrexed[(62.12±0.89)% vs.(21.56±0.67)% and(40.51±0.83)%, both P<0.05].
CONCLUSIONTS siRNA can partly suppress the expression of TS gene in LOVO cells, inhibit cell proliferation, promote cell apoptosis and enhance cell sensitivity to apoptosis induced by nolatrexd.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; enzymology ; pathology ; Humans ; Quinazolines ; pharmacology ; RNA Interference ; Thymidylate Synthase ; genetics ; metabolism