1.Optic nerve injury and tissue engineering technology
Chuanjie WANG ; Xiuhua YUAN ; Yingyu ZHAO
Chinese Journal of Tissue Engineering Research 2007;0(42):-
Optic nerve is composed by the axonal of retinal ganglion cells,there are no Schwann cells around it,and optic nerve belongs to the central nervous,so it cannot regenerate when injuried. In 1985 So and Aguayo have made a success in peripheral nerve retinal transplant,which completely changes the concept that optic nerve injury cannot regenarate. Currently nerve injury recovry has following several methods: use of neurotrophic factor,such as nerve growth factor,ciliary neurotrophic factor and brain-derived neurotrophic factor,etc. These factors could promote the regeneration and repair of retinal ganglion cells; treatment of gene correction and gene regulation,by changing the expression of host gene,reduce the effect due to the disease,slow the progress of the disease course or provide protection against the disease; neural stem cell transplantatin and tissue-engineered Schwann cell transplantion. But these treatments are still at the stage of animal experiment,how timely and effctively reduce the apoptosis and increase the survival rate of ganglion cells,even further promote nerve regeneration and recovery,there is still not a perfect method so far.
2.Effects of low frequency electric stimulation at bilateral mastoid processes on focal cerebral ischemia-reperfusion in rats
Jihong ZHOU ; Zhiqiang ZHANG ; Xiuhua YUAN
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(08):-
Objective To study the protective effects of low frequency electric stimulation at bilateral mastoid processes on the focal cerebral lesion induced by ischemia-reperfusion in rats. Methods Focal cerebral ischemia in rats was produced by 2 hours of the middle cerebral artery occlusion (MCAO) and 24 hours reperfusion. The neurological deficits scale was used to evaluate and screen the rats for tests. The cerebral water content was determined by wet/dry weighing method. The infarct volume was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining technique and image analysis instrument. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in ischemic tissue of brain were analyzed. Results Compared with the control group, the severity of cerebral edema in the treatment group, apart from the emergency group,was lessened(P
3.Inhibition effects of sulforaphane-induced autophagy flux to cell proliferation on ex vivo human capsular bags
Hanruo, LIU ; Bowei, YUAN ; Ying, AN ; Xiuhua, WAN
Chinese Journal of Experimental Ophthalmology 2017;35(3):226-232
Background Sulforaphane (SFN) is an effective chemopreventive agent and can regulate the biological molecular mechanisms to inhibit the overgrowth of cells.Autophagy is a biological process of maintaining cellular internal environment.Understanding the affection of SFN to biological behavior of human lens epithelial cells (LECs) and the association of SFN with autophagy is helpful for the prevention and target treatment of posterior capsule opacification (PCO).Objective This study was to investigate the eradication effeccts of SFN on residual lens cell population in vitro posterior capsule opacification (PCO) model and evaluate the mechanism of SFN-induced cell death.Methods In vitro human capsular bag models were generated from fresh donor eyes by phacoemulsification and were cultured in EMEM containing 2% fetal bovine serum (FBS).Different concentrations of SFN (0,1,10 and 100 μ mol) were added in the medium for 30 days respectively according to grouping,and the growth of LECs was observed by optical microscope and immunofluorescence technique.FHL124,a human LEC line,was cultured with EMEM containing 5% FBS and divided into 0,1,10,30 and 100 μmol SFN groups.Lactate dehydrogenase (LDH) release rate in the medium was detected to evaluate cell damage/death.The migration of the cells on capsular bags was assessed by scratch test.The ultrastructure and number of autophagosomes were examined under the transmission electron microscope.The expression of LC3 in the cells were detected using Western blot in the presence or absence of autophagy inhibitors.Results The cell coverage rates on the capsular bags were significantly lower in the 10 and 100 μ mol/L SFN groups than those in the 0 and 1 μmol/L SFN groups,with a statistically significant difference among the groups (F =48.57,P < 0.01).Immunofluorescence showed that the density of F-actin-and Vimentin-positive cells was evidently decreased in the 10 and 100 μmol/L SFN groups compared with 0 and 1 μ mol/L SFN groups.The releasing levels of LDH (absorbancy) were 0.19± 0.03,0.39±0.06,0.56±0.07,0.68±0.08 and 0.89±0.09 in the 0,1,10,30 and 100 μ mol/L SFN groups,respectively,and the releasing level of LDH was gradually increased in the 10 and 100 μ mol/L SFN groups in comparison with the 1 μmol/LSFN group (all at P<0.01).With the increase of SFN concentration,the reduction rate of scratched area decreased with the increase of SFN concentration,and the decrease of scratch area was significantly lower than that of adjacent low mass concentration group and the differences were statistically significant (P<0.05).The relative expressions of LC3-Ⅱ protein were 0.423±0.003,14.543±0.024,0.668±0.024 and 0.576±0.056 in the blank control group,SFN group,SFN + 3-MA group and 3-MA group,respectively,and the relative expressions of LC3-Ⅱ protein were significantly lower in the SFN+3-MA group and 3-MA group than those in the SFN group (all at P<0.01).The number of autophagosomes was 4.07±0.32,4.13±0.34,9.21 ±0.53 and 21.02± 1.34 in the blank control group,and 1,10,100 μmol/L SFN groups,and the number of autophagosomes in the 10 and 100 μ mol/L SFN groups was significantly higher than that in the blank control group and 1 μmol/L SFN group (all at P<0.01).Conclusions SFN mediates LECs death by promoting autophagy in ex vivo capsular bags,and SFN may be a novel agent of potential chemopreventive and target treatment for PCO.
4.Advances in Electric Therapies for Peripheral Nerve Injury (review)
Weiwei LI ; Xiuhua YUAN ; Jian YANG
Chinese Journal of Rehabilitation Theory and Practice 2010;16(7):640-641
In recent years,the repair and regeneration of peripheral nerve injury has developed. This paper reviewed the researches of peripheral nerve injury repairing from the low, medium, and high frequency electric therapy. The electric therapies should be selected with the pathology and regeneration of the injured peripheral nerve. The mechanism need further study.
5.Effects of low dose of hydrocortisone on circulating thymus-dependent lymphocyte apoptosis in patients with septic shock
Xiuhua LI ; Yongming CHEN ; Liang HONG ; Shoutao YUAN ; Shuyun ZHENG
Chinese Journal of Emergency Medicine 2010;19(5):528-532
Objective To investigate the effects of low-dose of hydrocortisone on circulating thymus-dependent lymphocyte (T lymphocyte) apoptosis in patients with septic shock. Method fifty-seven patients with septic shock admitted into ICU from January 2006 to January 2009 were prospectively randomized (random number) to treatment group and control group. Another 20 healthy volunteers and 18 patients with sepsis alone were included as external control groups.The patients of treatment group and control group were treated with low-dose of hydrocortisone and placebo,respectively. Samples of peripheral blood were taken from healthy volunteers and patients 0 hr,24 hrs,48 hrs,72 hrs and 168 hrs after onset of the disease to determine the circulating T lymphocyte apoptosis by using the assays of Annexin V and flow cytometry. Least significant difference t -test was used for multiple comparisons. Results The percentage of Annexin V-positive CD4+ T lymphocytes in the primary stage was (11.01 +4.52)% in septic shock patients, (4.41 + 1.45)% in healthy volunteers, and (7.87 + 3. 82)% in patients with sepsis alone. And in the initial setting, the percentage of Annexin V-positive CD4+ T lymphocytes in the septic shock patients was higher than that in healthy volunteers ( P < 0.05) and in patients with sepsis alone ( P < 0.05). The percentage of Annexin V-positive CD8 + T lymphocytes at the beginning was (11.33+19.62)% in septic shock patients, (9.62+8.32)% in healthy volunteers, and (13.09+ 15.84)% in patients with sepsis alone (P > 0.05 between three groups). The percentages of Annexin V-positive CD4+ T lymphocytes in control group after 24 his, 48 hrs and 72 hrs were(13.51+6.85)%, (19.39 + 6.63)% and (15.33+ 6.21)%, respectively. And the percentages of Annexin V-positive CD4+ T lymphocytes in treatment patients after 24 hrs, 48 hrs and 72 hrs were (17.4 + 7.21)%, (22.61 + 5.64)%, and (25.73 + 6.91)%, respectively. The percentage of Annexin V-positive CD4+ T lymphocytes in septic shock patients was higher than that in control groups ( P < 0.05). The percentages of Annexin V-positive CD8+ T lymphocytes in control group after 24 hrs, 48 hrs and 72 hrs were (11.49+ 11.73)%, (12.74+ 10.39)% and (13.28+ 16.6)%, respectively, and in the treatment group, those were (9.49 + 8.9)%, (15.32+18.17)% and (13.68+16.84)%, respectively (P >0.05 between two groups). In the meantime, the percentages of Annexin V-positive CDS'1' T lymphocytes in control group and in treatment group were (12.72+ 19.69)% and (13.88 + 13.28)%, respectively (P >0.05). Conclusions Low-dose of hydrocortisone could induce CD4+ T lymphocyte apoptosis and has no effects on CD8+ T lymphocyte apoptosis when it is used to treat septic shock.
6.Consumption Analysis of Oral Hypoglycemic Agents of 32 Hospitals in Wuhan Area during 2010-2012
Yongfang LEI ; Xiping LI ; Weigang YUAN ; Xiuhua REN
China Pharmacist 2014;(2):296-300
Objective:To analyze the utilization and tendency of oral hypoglycemic agents in some hospitals of Wuhan from 2010 to 2012. Methods:The relative data of oral hypoglycemic agents used in 32 hospitals of Wuhan area during 2010-2012 were analyzed using consumption sum, DDDs and defined daily dose as the indices. Results:The consumption sum of oral hypoglycemic agents was increased year after year;acarbose and gliclazide were widely used and occupied front places among the drugs. However, the percent-age of traditional Chinese medicine used for decreasing blood sugar was declined. Conclusion:During 2010-2012, the DDDs value and the consumption sum of oral hypoglycemic agents in Wuhan area are stable and normal, and the application will be further developed.
7.Experimental study of iontophoresis-mediated corneal collagen crosslinking for rabbit Staphylococcus aureus keratitis
Yue, YUAN ; Shaobin, ZHANG ; Yuqiang LIU ; Xiuhua LIU ; Sheng, WANG
Chinese Journal of Experimental Ophthalmology 2014;32(7):607-612
Background It is determined that riboflavin/ultraviolet A (UVA)-induced corneal collagen crosslinking is able to increase resistance of cornea against enzymatic digestion and has antimicrobial efficacy for various kinds of bacteria in vitro.However,its in vivo study is less now.Objective This study aimed to evaluate the efficacy of iontophoresis-mediated corneal collagen crosslinking combined with or without drugs for Staphylococcus aureus keratitis.Methods Bacterial keratitis models were induced by the interstromaly injection of Staphylococcus aureus suspension with concentration 2× 109/ml in the right eyes of 40 rabbits,and then the rabbits were randomly classified into the model group,gatifloxacin eye drops group,riboflavin/UVA corneal crosslinking group and drugs+ crosslinking group.The smearing of corneal surface was performed for the identification of bacteria 24 hours after injection.Iontophoresis-mediated riboflavin/UVA crosslinking was applied on the eyes of the riboflavin/UVA corneal crosslinking group and drugs+crosslinking group,and gatifloxacin eye drops was topically used 7 times per day on the eyes of the gatifloxacin eye drops group and drugs+crosslinking group.The corneal inflammation was examined and graded under the slit lamp biomicroscope before and after treatment.Ocular anterior segment optical coherence tomography(AS-OCT),corneal histopathology and ultrastructure were examined 14 days after treatment.The living environment of the experimental animals was maintained at 21 ℃ with a 12-hour light and dark cycle.Animals used in this study were treated in accordance with the Weifang Medical College Animal Experimentation Ethic Committee (AEEC) guidelines.The study protocol was approved by the AEEC.Results Corneal inflammation and ulcer were observed,but no significant difference was found in the inflammatory grade among the 4 groups 24 hours after injection (x2=0.293,P>0.05).In the 14th day after injection,the corneal ulcer area was smaller and corneal edema was milder in the drugs+crosslinking group compared with the model group,gatifloxacin eye drops group and riboflavin/ UVA corneal crosslinking group,showing a significant difference in the inflammatory grade among them (x2 =38.710,P<0.001).The cornea thickness values of ulcer zone were (428.1 ± 146.2) μm on the 14th postinjected day in the drugs+crosslinking group,which was evidently higher than those in the model group,gatifloxacin eye drops group and riboflavin/UVA corneal crosslinking group,with a significant difference among the 4 groups (F =8.310,P<0.001).A lower degree of destruction of cornea collagen and less inflammatory cells were seen in the cornea tissue of the drugs+ crosslinking group by haematoxylin and eosin staining in comparison with other 3 groups,and normal keratocytes were much more in the drugs + crosslinking group than those in other treated groups.Conclusions Iontophoresismediated corneal collagen crosslinking can alleviate Staphylococcus aureus keratitis.The combination of crosslinking with drugs has a better effectiveness than the administration of gatifloxacin eye drops only or riboflavin/UVA corneal crosslinking only.
8.Effect of Ultrashort Wave Therapy on Motor Conduction Velocity and Expression of Vascular Endothelial Growth Factor of Damaged Movement Neurons in Rats with Sciatic Nerve Injury
Weiwei LI ; Xiuhua YUAN ; Lixin ZHANG ; Jian YANG
Chinese Journal of Rehabilitation Theory and Practice 2010;16(8):744-747
Objective To investigate the effect of ultrashort wave therapy on motor conduction velocity (MCV) and expression of vascular endothelial growth factor (VEGF) in rats with sciatic nerve injury.MethodsThe models of peripheral nerve injury were established by forceps clip of sciatic nerve and then a total of 60 SD rats were divided randomly into three groups, including experimental group (n=24), control group (n=24) and sham operation group(n=12). The rats of experimental group were treated by ultrashort wave therapy after operation. The injured sciatic nerve and spinal cord in waist were sampled at the 1st,2nd,4th,6th week respectively after operation and were observed by electrophysiology and immunohistochemistry.ResultsAfter operation,the MCV of sciatic nerve in both experimental group rats and control group rats is zero in 1st week. Since 2nd week,the MCV of sciatic nerve in rats of both two groups began to arise and the MCV of injuried sciatic nerve in rats of experimental group was higher than that in the control group (P<0.05). In injured spinal cord motoneuron,the Integrated Optical Density(IOD) of VEGF in experimental group was higher than that in the control group (P<0.05).ConclusionUltrashort wave therapy could increase the value of MCV of sciatic nerve and the expression of VEGF in spinal cord in rats,and so it could protect the injured peripheral nerve.
9.Effects of ultra-shortwave irradiation on the expression of the apoptosis-related genes Bcl-2 and Bax in the lens epithelial cells of the eye
Zhongli WANG ; Xiuhua YUAN ; Yangyang LIN ; Xudong GU ; Yunhai YAO ; Jing WANG ; Jianming FU
Chinese Journal of Physical Medicine and Rehabilitation 2014;36(12):913-917
Objective To observe the effects of low-dose ultra-shortwaves radiationon the expression of apoptosis-related genes Bcl-2 and Bax in lens epithelial cells(LECs) of rats.Methods Thirty-six Wistar rats were divided into a normal group(n=8 eyes),a control group(n=32 eyes) and an experiment group(n=32 eyes) randomly.Both eyes of each rat in the experiment group were exposed to a low-dose ultra-shortwave radiation for 7min,once a day until the end of experiment,while the control groups were not.Rats were executed after 1 w,3w,6w and 9w.Both eyes were resected,and lens were separated under a microscope and sectioned in paraffin.After Streptavidin-Biotin Complex (SABC) staining,Bcl-2 and Bax expression in the LECs were detected using an image analysis technique,and the data were analyzed using the software of SPSS 13.0.Results The average integral optical density (AIOD) of Bcl-2 expression in the experiment group was (0.391 ± 0.014) after 1 week,(0.4470.006) after2,(0.417 ±0.011) after3 and (0.275 ±0.007) after4 weeks.The corresponding AIODs for Bax expression were (0.180 ±0.015),(0.155 ± 0.007),(0.167 ± 0.003) and (0.251 ± 0.016) respectively.After l w of daily radiation,no significant difference was found in the expressions of Bcl-2 and Bax protein between the experimental and the other 2 groups(P > 0.05).Three weeks after the radiation,however,the expression of Bcl-2 protein in the experimental group was significantly higher than the control groups(P<0.001),with that of Bax being significantly lower(P<0.001).However,the expressions of Bcl-2 protein was significantly down-regulated and the expression of Bax protein was significantly up-regulated 6w later compared with those after3 w of exposure in the experiment group(P<0.001) After9w,in the experimental group,the expressions of Bcl-2 protein was obviously down-regulated and significantly lower than the control group.Moreover,the expression of Bax protein increased significantly compared to the control group (P<0.01).And there was no significant difference between the control and normal group(P>0.05) in the expression of Bcl-2 and Bax.Conclusions Low doses of ultra-shortwave radiation may protect LECs against apoptosis for a few days,but they may cause damage with repeated exposure.
10.The effect of 810 nm low-power Ga-Al-As laser treatment on the regeneration of a damaged optic nerve
Yangyang LIN ; Xiuhua YUAN ; Fangqian ZHOU ; Zhongli WANG ; Miaomiao LIU ; Zhenyuan LIN
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(9):662-667
Objective To determine whether or not 810 nm low power Ga-Al-As laser treatment can stimulate the regeneration of damaged optic nerves by measuring the expression of growth associated protein 43 ( GAP-43 )and flash-visual evoked potential (F-VEP). Methods Eighty-eight Wistar rats weighing (180-220) g were randomly divided into a laser therapy group with 40 rats,an injury group with 32 rats,and a normal control group with 16 rats.Each group was subdivided into 1st,3rd,6th and 9th week subgroups.A standardized crushing of the optic nerve was applied to make the model.After this,the laser therapy group was treated for 3 minutes daily at 60 mW applied transcutaneously to a 5 mm diameter spot on the injured optic nerve.The injury and normal control groups received the same treatment with no laser output.The expression of GAP-43 was detected by immunohistochemistry and RT-PCR after 1,3,6 and 9 weeks of treatment.F-VEP was measured pre-injury,immediately after injury and 1,3,6 and 9 weeks post injury. Results After the optic nerve was injured,obvious changes in F-VEP were detected,including significantly prolonged latencies of N1,P1 and N2 waves.The latency increased immediately after the optic nerve injured,and then recovered,but after 1 and 3 weeks the latency was still prolonged.There was significant recovery from the 3rd to the 9th week.In the laser therapy group,the peak latencies of the N1,P1 and N2 waves were also prolonged,but the changes were less than those in the injury group.Expression of GAP-43 was hardly detectable in normal retinas and optic nerves.GAP-43 had its highest expression level at 1 week post-injury,and then decreased.At the 1st,3rd and 6th week post-injury,the expression of GAP-43 in the laser therapy group was significantly higher than in the injury group.GAP-43 mRNA content in the retina showed the same tendency as GAP-43 protein. Conclusion A 810 nm low power Ga-Al-As laser can promote neural repair and axonal regeneration after optic nerve injury.