1.Detection of ?-nerve growth factor secreted by hair follicle bulge cell cultured in vitro
Journal of Third Military Medical University 2003;0(09):-
Objective To detect ?-nerve growth factor(?NGF) secreted by hair follicle bulge cells cultured in vitro qualitatively and quantitively and search the relationship between ?-NGF and bulge cells growth condition.Methods The primary tissues from the labial part and around the barbell in inbred Wistar rats aged 6-8 d were stripped by micromanipulative technique and cultured.The ultrastructure of primary bulge cells was observed under transmission electron microscope(TEM).The secretion of ?-NGF was determined by ELISA and immunocytochemistry.Results Primary bulge cells were cultured in vitro successfully.?-NGF was strongly expressed in the plasma of cultured bulge cells detected by ICC.The secretion of NGF detected by ELISA was regularly correlated with the characteristic of primary cultured bulge cells.Conclusion Primary bulge cells secreted the highest ?-NGF when bulge cells grew into peak phase.The expression of ?-NGF must have some necessary relationships with hair follicle bulge cells.
2.Inhibitory effect of iNOS inhibitor on the apoptosis of rat abdominal aortic aneurysm smooth muscle cells
Chinese Journal of Current Advances in General Surgery 2017;20(3):178-181
Objective:To explore the establishment of an animal model of abdominal aortic aneurysm(AAA),and elaborate the role of iNOS inhibitor in the smooth muscle apoptosis of abdominal aortio aneurysm in rats,to find a new theoretical basis for the clinical treatment of small drugs AAA.Methods:SD rats underwent intra-aortic elastase (25U/mL) perfusion to induce AAAs,the positive control group (30) and experiment group (30) use elastase perfusion while the negative control group(30) gives the saline perfusion.After operation the positive and negative control groups were treated with intraperitoneal injections of saline,experimental group injects the iNOS inhibitor Aminoguanidi hydrochloride;Postoperative second,7,and 14 days,The NO content in the serum and specimen of abdominal aortic aneurysm was detected by iNOS Immuno histochemistry and Terminal Transferase-mediated dUTP Nick End-Labeling (TUNEL) to evaluate distribution of smooth muscle apoptosis in abdominal aortic aneurysm.Results:Underwent intra-aortic elastase perfusion to induce AAAs have a high-success-rate.Rate of AAA formation in positive control group 10%,60%,80%,respectively.The treatment group was 0%,10%,20%,and the negative control group was not formed.The treatment group and the negative control group were lower than the positive control group,there were significant differences.In the positive control group,NO content increased gradually from second days,7 days to reach the peak and maintained at a higher level,the treatment group serum NO content was lower than the other two groups,there was significant difference (P<0.05),iNOS was strong expression in the positive control group,in the other two groups of mild expression.TUNEL results showed that a lot of apoptotic cells in the positive control group,after 7 days showed a significant increase trend,to observe the end (2 weeks) gradually increased.,The positive control group was higher than the negative control group and the negative control group,there was significant difference(P<0.05).Conclusion:iNOS inhibitors significantly decreased the content of NO in serum,reduced the apoptosis of smooth muscle cells,and inhibited the formation of abdominal aortic aneurysm.To provide theoretical basis for clinical application of iNOS inhibitors in the treatment and control of AAA.
4.Effects of Exercise and Aloe on Serum Antioxidant Enzyme Activity in Diabetic Rats
Wentao LIN ; Sen HUANG ; Xiquan WENG
Chinese Journal of Sports Medicine 1982;0(01):-
Objective To investigate the effect of exercise and a loe on serum antioxidant enzyme activity in diabetic rats. Methods Adult male Streptozotocin-diabetic rats were used as research subject. Afte r exercise, aloe and exercise combines aloe treated, the changes in SOD, GSH-Px, CAT activity, MDA contents, blood glucose and insulin were measured. R esults The level of SOD, GSH-Px, CAT activity and insulin of treated diabetic groups were significant higher (P
5. Preparation of gastrodin starch microsphere and its nasal mucoadhesion and in vitro release characteristics
Chinese Traditional and Herbal Drugs 2016;47(4):585-590
Objective: To determine the prescription technology of gastrodin starch microsphere and investigate its nasal mucoadhesion and in vitro release characteristics. Methods: Gastrodin starch microspheres were prepared by compound emulsion crosslinking method. According to the particle diameter, drug loading efficiency (DLE), and entrapment efficiency (EE), the best prescription technology was selected by using single-factor investigation and uniform design. Using toad palate mucosa as model and average residence time as indicator, mucoadhesion of gastrodin starch microsphere was evaluated. Using gastrodin API as a control, paddle method was applied to in vitro release test of gastrodin starch microspheres. The content of gastrodin was determined to calculate the cumulative release percentage. In addition, the curve of drug release in vitro was fitted with different release model to analyze the in vitro release characteristics of gastrodin starch microsphere in nasal cavity, synthetically. Results: The optimum prescription and preparation technology of gastrodin starch microsphere were as follows: gastrodin 2.0 g, starch 4.5 g, liquid paraffin 100.0 mL, Span80 3.5 g, ECH 5.1 mL, preparation temperature 40℃, and rotational speed 1000 r/min. The particle diameter of gastrodin starch microsphere was (47.69±1.92) μm, the DLE and EE of microsphere were (9.78±0.70)% and (35.72±3.28)%, respectively. It was about (176.92±23.25) s that in adhesive powder resided in nasal cavity, which translated into human nasal residence time was just 20-30 min, while the average residence time of gastrodin starch microspheres was extended to (944.33±68.29) s, translated into human nasal residence time was about 3 h. The cumulated release percent of gastrodin starch microspheres was more than 90% in 3 h. Compared with other in vitro release models, Weibull model was the fittest model to gastrodin starch microspheres, the t50 of gastrodin starch microspheres was 40.08 min, and t90 was 245.73 min. Conclusion: Gastrodin starch microspheres prepared with optimum prescription technology have uniform particle diameter, high DLE and EE. Microspheres have good mucoadhesion and sustained release, ensure that gastrodin release gently and completely during the nasal retention period.
6.Effects of cognitive impairment in hippocampus on glucose and lipid metabolism and its relations with gastrointestinal motility in rats
Lin YE ; Sen DU ; Lin ZHU ; Chunbo XIA
The Journal of Practical Medicine 2015;(17):2789-2792
Objective To investigate the effects of cognitive impairment in hippocampus on glucose and lipid metabolism, and its relations with gastrointestinal motility. Methods The Aβ1-42 was injected into the hippocampus of rats. Levels of glucose and lipid were detected. The changes of gastrointestinal motility were detected by the type-B ultrasonic and the ink-pushing experiments. Hippocampal neurons apoptosis was detected by the TUNEL assay. Results In the experimental group, FPG, TG, TC, LDL were (7.92 ± 0.29) mmol/L, (2.24 ± 0.12) mmol/L, (4.67 ± 0.12) mmol/L, (2.41 ± 0.12) mmol/L, respectively, with significant differences among these three groups (P < 0.05). Compared with the sham group and the control group, the number of bowel movements per unit time (2.13 ± 0.83) times, gastric emptying rate (44.35 ± 7.53) % and the small intestinal propulsion rate (57.60 ± 7.82)%in the experimental group were significantly decreased (P<0.05). The experimental hippocampal neuronal apoptosis index was an average of (64.98 ± 3.70)%, which was significantly higher than that in the sham group and the control group (P < 0.05). Conclusion Hippocampal cognitive impairment can elevate the blood lipid level, which may be associated with the hippocampal neuronal apoptosis and the gastrointestinal motility disorders.
7.Influence of beta nerve growth factor in vibrissa follicle bulge region on the burn wound healing of neonatal rats
Jun ZENG ; Dengbang CHEN ; Lüxia DAI ; Sen LIN
Chinese Journal of Tissue Engineering Research 2010;14(11):1987-1991
BACKGROUND:The surroundings stimulation,such as wound and chemical injury,will result in changes of hair follicle stem cells(HFSC).Up to date,few articles indicate the effecl of burn on HFSCs.OBJECTIVE:To investigate the changes of β-nerve growth factor(β-NGF)level in vibrissa follicle bulge region and its effect on the healing of neonatal rats.METHODS:A total of 30 neonatal rats were selected to prepare burn models by giving 90℃ boiled tap water for 3 seconds,and vibrissa follicle bulge was separated at 12,24,36,48 and 60 hours pOsloperatiVeIy.The remained 6 neonatal rats were served as controls.Total protein concentration was measured by Brandford method The expression of β-NGF was detected by ELISA.Meanwhile the tissues in each time points were collected for frozen section and immunohistochemistry.RESULTS AND CONCLUSION:Among the tested total protein.the expression of the β-NGF increased after 36 hours.reached a peak at 60 hours after burn.The identical changes were found in tissue slices.The image analysis demonstrated that the β-NGF protein detection was coincident with β-NGF expression in tissues During 12 hours to 60 hours after burn,the expression of the β-NGF in the bulge area peaked at 60 hours;simultaneously,the repairing effect of healing also reached a peak.Bulge area is a nest of HFSC,the enhanced expression of β-NGF may have some positive significance to the differentiation of the HFSC and tissue repair.
8.Relationship of human microvascular endothelial cell activation and adenosine level under the hypoxia
Liqianyu, AI ; Chanjuan, HUANG ; Chen, CHEN ; Sen, LIN ; Jian, YE
Chinese Journal of Experimental Ophthalmology 2017;35(1):26-31
Background Retinal hypoxia is one of primary causes of retinal neovascularization,and its mechanism is research hot topic.Studies showed that hypoxia stimulates the activation of many trancripation factors and vascular endothelial cells,which leads to angiogenesis.Besides to the vascular endothelial growth factor,the effect of adenosine on angiogenesis is increasingly concerned.Objective This study was to invastigate the relationship of biological behaviour of human microvascular endothelial cells (HMEC-1) under the hypoxia and adenosine,and to explore the effect of adenosine on angiogenesis under hypoxia.Methods HMEC-1 cell line was cultured in vitro,and the cells were divided into normoxia group and hypoxia group.The cells in the normoxia group were cultured under the 5% CO2 environment,and those in the hypoxia group were cultured under the 1% O2,94% N2 and 5% CO2 environment.The proliferation ability and percentage of the cells were assayed by cell counting kit-8 (CCK8) and EDU.The migration number and invasive number of the cells were detected by transwell chamber.The expressions of CD39 and CD73 proteins in the cells were tested by Western blot and immunofluorescence technique,and the level of adenosine was measured by high performance liquid chromatography (HPLC).Results The proliferation values (absobancy) were 0.715-±0.067 and 0.821 ±0.056 in the normoxia group and the hypoxia group in 12 hours after culture,and those in 24 hours were 0.946±0.028 and 0.998±0.028,showing significant increase in the hypoxia group compared with the normoxia group (t12h =3.805,t24h =3.222,all at P < 0.01).The precentage of the proliferation in the hypoxia group was evidently higher than that in the normoxia group (t =-6.868,P<0.01).The number of the cell migration and invasion in 24 hours after culture was 185.3 ± 10.594 and 74.2± 10.741 respectively in the normoxia group,and that in the hypoxia group was 300.7±22.853 and 107.5±7.007,with significant differences between the two groups (t=-12.124,-6.367,both at P<0.01).The expression levels of CD39 and CD73 proteins in the hypoxia group were significantly higher than those in the normoxia group in bothl2 hours and 24 hours after culture(all at P<0.05),and the adenosine content in the cells is gradually increased in 2,6,12,24 and 36 hours after culture,with the highest content in 36 hours.The adenosine content was significantly higher in the hypoxia group than in the normoxia group at various time points (t2h =2.469,P =0.017;t6h =5.442,P<0.001;t12h =3.841,P<0.001;t24h =4.458,P<0.001;t36h =2.757,P =0.008;t48h =3.319,P =0.002).Conclusions Compared with the normoxia group,the proliferation,migration and invasion abilities of HMEC-1 are stronger,meanwhile,the expression of CD39 and CD73 as well as the adenosine level in the cells are all increased under the hypoxic condition.It is suggested that the activation of human microvascular endothelial cells might be significantly related to the level of adenosine and its key enzymes.
9.The detection of treatment of bile duct system stones with Qingdanyin and ESWL
Sen MA ; Qun LIN ; Nan LI ; Meihong LI
Chinese Journal of Primary Medicine and Pharmacy 2005;0(11):-
Objective To evaluate the curative effect on chdecytolithiasis with Chinese traditional patent formulation Qingdanyin and ESWL. Methods From January 2000 to January 2003,332 patients with gallstones were treated with Qingdanyin and ESWL, compared with 141 patients with routine ESWL. Results The 2-week clean rate(36.14%),one month clean rate(58. 58%),3-month clean rate(68. 98%) ,6-month clean rate(86.75%) of gallstones in the Qingdanyin and ESWL group were much higher than those in routine ESWL patients(14. 92%, 21.02%,35.93%,47.11%,respectively)(P
10.Effects of oral rehydration On hemodynamics and microcirculatory perfusion in dogs with fatal hemorrhagic shock
Sen HU ; Lin LI ; Jingyuan HOU ; Ruichen WANG
Chinese Journal of Anesthesiology 2010;30(4):448-451
Objective To investigate the effect of oral rehydration on hemedynamies and mierocirculatory perfusion in dogs with fatal hemorrhagic shock.Methods Twenty male Beagle dogs 16-20 months old weighing 8-12 ks were subjected to a loss of 40% of the total blood volume,then divided into 3 groups:no rehydration group (group NR,n=8),oral rehydration group(group OR,n=6)and intravenous rehydration group(group IR,n=6).Group NR received no treatment within 24 h after blood-letting.Group IR and OR were given glucose-electrolyte solution (GES) either by gastric tube or by intravenous infusion 3 times volume of the blood loss immediately after the establishment of the model.Then the lactated Ringer's solution,glucose saline and compound amino acid(2 times volume of the blood loss)were started to be given to supplement the physiological consumption from 24 h after blood-letting in each group.The MAP,cardiac index(CI),systemic vascular resistance (SVR),dp/dtmax,and intestinal mucoflal blood flow (IMBF) were determined before blood-letting(T0,baseline) and 2 h (T1),4 h(T2),8 h(T3),24 h(T4),48 h(T5) and 72 h(T6)after blood-letting.The fatality rate within 72 h after blood-letting and urinary output were calculated.Results The fatality rates were 63%,33%and O in group NR, OR and IR respectively, which showed significant difference between the groups (P < 0.05).Compared with the baseline values at To, MAP, CI and dp/dtmax were significantly decreased at T1-6, in group NR,at T1-5 in group OR and at T1-4 in group IR, and SVR was significantly increased, while IMBF decreased at each time point after blood-letting in the three groups ( P <0.05), but no significant change was found in MAP, CI and dp/dtmax at T6 in group IR and OR (P>0.05). MAP, CI, dp/dtmax , IMBF and urinary output were significantly higher, while SVR was significantly lower in group OR and IR than in group NR ( P < 0.05). MAP, CI,dp/dtmax, IMBF and urinary output were signiflcandy lower, while SVR was significantly higher in group OR than in group IR ( P < 0. 05). Conclusion Oral administration of GES 3 times volume of the blood loss within 24 h after fatal hemorrhagic shock can obviously improve the hemodynamics and microcirculatory perfusion, then improve the survival state and have obvious resuscitation efficacy.