1.Characteristics of mouse embryonic hepatic stem cells in different embryo durations
Bigang WU ; Jing CHANG ; Xiaogang ZHANG
Chinese Journal of Tissue Engineering Research 2013;(23):4279-4285
10.3969/j.issn.2095-4344.2013.23.014
2.Mouse embryonic hepatic stem cells differentiate into cardiomyocyte-like cells
Bigang WU ; Jing CHANG ; Xiaogang ZHANG
Chinese Journal of Tissue Engineering Research 2013;(32):5813-5818
BACKGROUND:In recent years, embryonic hepatic stem cel s have attracted more attention, but there are few reports on the potential of embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s as wel as the related differentiation conditions. OBJECTIVE:To investigate the moderate condition to induce mice embryonic hepatic stem cel s to differentiate into cardiomyocyte-like cel s in vitro with chemical reagents. METHODS:Dimethylsulfoxide in combination with 5-azacytidine with different concentrations and time were used to induce the embryonic hepatic stem cel s of 13.5 days mice and to observe the differentiation effect. RESULTS AND CONCLUSION:Under in vitro conditions, 0.8%dimethylsulfoxide+5μmol/L 5-azacytidine could induce the mouse embryonic hepatic stem cel s to express the specific markers of myocardial cel s, while increasing the concentration of the inducer and extending the induction time could not improve the induction efficacy.
3.Isolation, incubation and identification of mouse embryonic hepatic stem cells
Bigang WU ; Xiaogang ZHANG ; Jing CHANG
Chinese Journal of Tissue Engineering Research 2007;0(10):-
BACKGROUND: Fetal liver cells can have stronger abilities to proliferation and differentiation and lower immunogenicity compared to bone marrow stem cells. However, there are few studies on direct isolation and culture of embryonic hepatic stem cells (EHSCs). OBJECTIVE: To isolate and cultivate EHSCs in vitro and to identify their biological features. DESIGN, TIME AND SETTING: The cytology in vitro controlled study was performed at the Chongqing Key Laboratory of Neurology from March to June 2008. MATERIALS: A total of 9 SPF Kunming fetal mice aged 13.5 days were obtained from Animal Experimental Center of Chongqing Medical University. METHODS: Collagenase + EDTA digestion and differential adherence method were used to isolate EHSCs, which were then incubated at 2?108 /L. Cells were digested and passaged when 80%-90% cells were confluent. Using streptavidin-biotin-peroxidase complex technique, adhered cells following 5 days of incubation were labeled with various EHSC surface marker. MAIN OUTCOME MEASURES: Morphology, passage, amplification and surface marker surface of EHSCs were measured. RESULTS: The isolated EHSCs adhered to the culture plastic and presented pykno-round cells and distinct borderline 24 hours after cultivation in vitro. Cells grew spindle-shaped 3 days. After 7 days they grew like epithelium. Cell amplified speed following passage did not have significant changes. Cells still presented epithelium-like shape at the passage 5. The adhered cells at day 5 following primary incubation were positively for human stem cell factor receptor and alpha fetoprotein, and negatively for albumin and cytokeratin 19. CONCLUSION: EHSCs were positively for human stem cell factor receptor and alpha fetoprotein, and negatively for albumin and cytokeratin 19 in early primary culture. This indicated that the cultivated cells are proved to be primordial progenitor cells and still in undifferentiated early phase.
6.Applied anatomy and biomechanics of atlas pedicle screw implantation
Zhiqiang CHANG ; Pei ZHANG ; Yimin WU ; Pengfei LI
Chinese Journal of Tissue Engineering Research 2014;(26):4185-4189
BACKGROUND:A series of pathological changes were found in atlas, including atlas dislocation, atlanto-occipital junction instability and vertebral fractures. Atlantoaxial pedicle screw, nail and rod fixation or nail plate fixation is a novel repair method recently proposed, and has certain advantages. OBJECTIVE:To compare the repair effects of atlas pedicle screw on different types of atlas lesions. METHODS:A total of 42 fresh vertebrae were obtained from Department of Human Anatomy, Basic Medical School, Inner Mongolia Medical University. In accordance with bone thickness of sagittal vertebral artery groove bottom, vertebrae were divided into three types:type I (common type), type II (light variation), and type III (severe variation). Al samples were subjected to atlas pedicle screw fixation. RESULTS AND CONCLUSION:The screw pul out force in different bone densities of vertebrae was not the same. Vertebrae with great thickness and great bone density had big screw pul out force (P<0.05). However, the screw in different cortical bone of vertebrae had the same function. No compression on blood vessels or phenomenon affecting vertebral activity was visible. These results indicated that atlas pedicle screw implantation showed good safety, no side effects, did not affect the normal work and life of patients.
7.Evaluation of infrapopliteal occlusive disease with contrast-enhanced MR angiography in diabetic patients
Lei ZHANG ; Jian CHANG ; Yun WU ; Dongchun SHI ; Xiaoying ZHAO
Chinese Journal of Radiology 2014;48(8):664-669
Objective To evaluate the diagnostic accuracy of contrast-enhanced MR angiography (CE MRA) in infrapopliteal occlusive diseases of diabetic patients.Methods A total of 105 patients with known diabetes and peripheral vascular occlusive disease who underwent both CE MRA and DSA examnations were included in this study.They had no obvious stenosis or with stenosis of less than 75% in iliac,femoral,and popliteal arteries.Every infra-popliteal artery was anatomically divided into 9 vascular segments as tibiofibular trunk artery,proximal and distal anterior tibial artery,proximal and distal posterior tibial artery,proximal and distal peroneal artery,plantar and dorsalis pedis artery.There were 945 segments.The arterial stenosis was accessed with CE MRA and DSA respectively.The segments were scored in 5 categories as<30%,≥30% and<50%,≥50% and<75%,≥75% and<100%,and 100% according to stenostic degrees.The Kappa test was used to compare the diagnostic consistency of CE MRA and DSA.Taken DSA as a gold standard for reference,ROC curve was drawn to calculate the diagnostic accuracy of CE MRA in diagnosis of lower limb arterial disease.Results The ratio of statistically valid segments for both CE MRA and DSA were 97.7% (923/945) in 945 vessel segments of 105 patients,and 390 of 923 arterial segments were both indicated as occlusion by CE MRA and DSA.The diagnostic consistency for the segments was listed in the decreasing order as follows:proximal peroneal artery,proximal posterior tibial artery,distal posterior tibial artery,proximal anterior tibial artery,distal anterior tibial artery,tibiofibular trunk artery,dorsalis pedis artery,plantar artery,distal peroneal artery,and corresponding Kappa values were 0.88,0.86,0.84,0.84,0.81,0.77,0.75,0.75 and 0.73,P< 0.05.The AUC(area under ROC curve) of CE MRA was 0.893 with 95% confidence interval of 0.882 to 0.904.Conclusion CE MRA is an accurate imaging modality in the diagnosis of infrapopliteal occlusive disease s for diabetes.
8.Expression of GABA and glutamate vesicular transporter in depressed mice
Qiong WU ; Jingfang SONG ; Xiaoling ZHANG ; Hongyang CAI ; Quanzhong CHANG
Chongqing Medicine 2016;45(11):1470-1472
Objective To study the expression of vesicular GABA transporter and vesicular glutamate transporter 1 in de‐pression .Methods Mice was divided into control group and defeat group stochastically .By social defeat model and social avoidance , the defeat group was divided into two groups:susceptible group and unsusceptible group .Synaptic proteins were extracted respec‐tively from the 3 groups .We detected the expression abundance of VGAT and VGLUT1 by Western blot .Results Compared with the control group ,in susceptible group ,the residence time in the contact area was significantly reduced ,and the residence time in the corner area was significantly increased ,with statistical difference(P<0 .05) .In the prefrontal cortex and hippocampus ,Com‐pared with the control group and the unsusceptible group ,the expression levels of VGLUT1 and VGAT were increased in the sus‐ceptible group(P<0 .05) .there were no statistically significant in VGLUT1 and VGAT leveles between control group and the un‐susceptible group(P>0 .05) .In the striatum ,although the expression levels of VGAT and VGLUT1 were increased in susceptible group ,but in unsusceptible group ,the expression of these proteins also increased significantly .Conclusion The prefrontal cortex and hippo‐campus excitability and inhibitory vesicle transport were changed in depression ,which may relate to the transcription disorder .
9.Value of P300 detection in evaluating cognitive impairment of patients with silent cerebral infarction and depression
Mingze CHANG ; Weiping ZHANG ; Yingxian ZHAO ; Haiqin WU ; Li XIANG
Chinese Journal of Tissue Engineering Research 2006;10(14):174-176
BACKGROUND: Cognitive impairments are often accompanied in patients with silent cerebral infarction (SCI) and depression, and there is still no reliable objective assessing method for the evaluation.OBJECTIVE: The purpose of this study was to probe into the value of auditory P300 in detecting the cognitive impairment in SCI patients with depression.DESIGN: A golden standard control study.SETTING: Department of Neurology, Xi 'an Central Hospital; Department of Neurology, the Second Hospital of Xi' an Jiao Tong University.PARTICIPANTS: Thirty-eight SCI patients were selected from the Outpatient Department of Neurology, the Second Hospital of Xi' an Jiao Tong University between January 2002 to May 2003, including 10 got dizziness,11 got poor concentration, 8 got headache, 5 got numbness of extremities and 4 got syncope. Among them, 26 were detected by CT and 12 by MRI.The patients'aged 45 to 63 years with mean age of (57.3±5.1) years, including 20 males and 18 females.METHODS: The existence of depressive symptoms was measured with the 17-item Hamilton depression scale (HAMD). In this scale, the total score ranged from 0 to 52, with higher score indicating more severe depressive symptoms, and the patient was considered as having depression if the HAMD score was ≥ 9. HAMD tests were administered by two psychiatrists,who were experienced in psychiatric disorders in stroke and other somatic diseases, and the two psychiatrists had a better coincidence (Kappa=0.86 -0.91). The auditory event related potential was detected with MYTO serial cerebra induced potential meter, while lying in a bed placed in a sound-attenuated room. Auditory stimuli were delivered binaurally through headphones. The subject was required to distinguish between the tones by noting the occurrence of the target and not to respond to the standard. All patients underwent a routine experimental test at first, and then a normal test was performed to obtain a better coincidence. Mini-mental state examination (MMSE) was conducted for the 38 patients by specialized training neurologist to assess the cognitive impairment, in which the total score ranged from 0 to 30, with lower score indicating greater impairment. The criteria of intellectual deterioration was defined on the basis of educational level:MMSE score of 17 for illiterate, 20 for primary school and 24 for middle school and above.MAIN OUTCOME MEASURES: The score of MMSE, latent period of P300 wave and amplitude of P300 wave were mainly observed.RESULTS: All the 38 cases were involved in the final statistical analyhave depressive symptoms (47%, depression group) and 20 had no depresindexes and score of MMSE between the SCI patients with and without depression: As compared with the SCI patients without depression, the P300 latent period was obviously prolonged, the P300 amplitude and MMSE score were markedly decreased in the SCI patients with depression [(383.3±31.1), (347.2±37.3) ms; (5.6±0.7), (9.9±1.6) μV; (15.2±1.1),and P300 indexes: In the SCI patients with depression, the P300 latent period had significant negative correlation with the MMSE score (r=-0.549, P < 0.05), the P300 amplitude had significant positive correlation with the MMSE score (r =0.525, P < 0.05). However, there was no correlation between the latent period and amplitude of P300 and the MMSE score in the SCI patients without depression (r=-0.349 to 0.065, P > 0.05).CONCLUSION: Depression and cognitive impairments may have the same pathophysiological basis in SCI patients. P300 can be used as a referential and objective method to analyze the cognitive function in SCI patients with depression.
10.Effect of ultraviolet irradiation dose and style on 25-hydroxyvitamin D and bone metabolism in growing rats
Wei ZHANG ; Liwen CHANG ; Shurong WANG ; Wei WU
Chinese Journal of Tissue Engineering Research 2009;13(46):9192-9196
BACKGROUND:Sunshine time in northem region is short in winter,the infants and young children are vulnerable to lack of vitamin D. Up-to-date textbooks and the guidelines formulated by Chinese Medical Association account that glass block ultraviolet and indoor exposure to human is meaningless. Early studies have shown that sunlight exposure through glass had meaning in rats,but it was difficult to accurately quantify,while outdoor exposure in rats is difficult to continue. This expedment uses B-band ultraviolet rays,which has the impact on vitamin D metabolism in some wavelengths,to facilitate further study.OBJECTIVE:To study the effect of ultraviolet B exposure in laboratory on 25-hydroxy vitamin D(25-OHD)level and bone metabolism in the serum of growing rats.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed in June 2007 in the ExperimantalAnimal Center of Chinese PLA General Hospital.MATERIALS:Forty male 21-day-old Wistar rats were employed in this study. Ultraviolet waves were sourced from an artificial UV light instrument(wavelength 280-350 μm,irradiation intensity 5.5μW/cm~2).METHODS:Forty Wistar rats were divided into four groups by random sampling table,exposed to different doses of ultraviolet Bwave. Direct exposure group:direct exposure for 20 minutes at an irradiation dose of 4.2 mJ/(cm~2·d);Indirect exposure 60 and 120 min groups:exposure through a single pane of glass for 60 and 120 minutes at an irradiation dose of 0.36 and 0.72 mJ/(cm~2·d).Control group:no exposure was given. For the direct exposure group,the lamp was placed above the cage. A 3-mm-thick pane of glass(common window glass) was placed underneath the lamp in the indirect exposure groups. Exposure groups were given irradiation for successive 20 days.MAIN OUTCOME MEASURES:Serum 25-OHD,bone alkaline phosphatase (BALP),and bone mineral density (BMD) were measured on day 21.RESULTS:25-OHD was significantly higher in all exposure groups compared with the control group (P<0.001),but there was no difference between the direct exposure group and two indirect exposure groups. BALP was significantly lower in the exposure groups than control group (P<0.001),it was also significantly lower in the 120-min indirect group than in the 60-min indirect group (P=0.022). There was a positive correlation between exposure dose and 25-OHD (r=0.555,P=0.002) and a negative correlation between exposure dose and BALP (r=0.595,P=0.001),also a negative correlation between 25-OHD and BALP (r=0.569,P=0.002),but there were no differences between groups for BMD. Exposure dose exhibited a threshold,serum 25-OHD and BALP no longer increased or decreased when it was 0.36 mJ/(cm~2·d).CONCLUSION:Midwave ultraviolet rays might affect serum 25-OHD and BALP levels in the growing rats through glass exposure,with no significant difference compared to direct exposure. The B-band ultraviolet exposure dose may play an important role in serum 25-OHD synthesis,but there is a threshold dose for the synthesis. Low-dose and prolonged exposure time also achieve threshold exposure.