Prevalence and risk factors of dry eye patients with corneal epithelial defects: A hospital-based retrospective study / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the characteristics and correlated risk factors of dry eye patients with corneal epithelial defects.METHODS: Outpatient medical records of dry eye patients with corneal epithelial defects at Peking University Third Hospital from July 2018 to June 2019 were retrospectively analyzed. The patients' data including sex, age, visit date, presence of comorbidities, and meteorological indicators at the same period were statistically analyzed.RESULTS: A total of 291 dry eye patients with corneal epithelial defects, of whom 75.3% were female, were retrospectively analyzed. Young patients aged 21-30 made up the most(26.5%), while the proportion of teenagers(<18 years, 5.8%)and the elderly(≥61 years, 17.2%)was low. However, as the largest proportion of this population, young and middle-aged patients tend to experience fewer visits(5.4±12.4). Spring and winter were the main seasons of complaints. The meteorological indicators at the same period including fine-particulate matter with a median aerometric diameter of less than 10μm(PM10), sulfur dioxide(SO2), nitrogen dioxide(NO2), and reduced average relative humidity were found significantly correlated with dry eye corneal epithelial defects(P<0.05). Conjunctivitis, cataracts, blurred vision, and trichiasis ranked the top four comorbidities.CONCLUSION: Dry eye corneal epithelial defects of young and female population cannot be ignored. PM10, SO2, NO2, and reduced humidity are found significantly correlated with dry eye corneal epithelial defects. For dry eye patients with conjunctivitis, cataracts, blurred vision, and trichiasis, more attention should be paid to their corneal conditions.

Analysis of fluorescence fundus angiography in patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To research and discuss the value and features of fluorescence fundus angiography(FFA)in patients with diabetic retinopathy. <p>METHODS: We selected 130 hospitalized diabetic patients suspected with diabetic retinopathy from January 2014 to February 2018 in our hospital. All patients underwent fundus photography and FFA examination. We analyzed the detected retinopathy. The reference was the clinical diagnosis, to calculate the sensitivity, specificity and accuracy of fundus photography and FFA in diagnosis of diabetic retinopathy and to compare the diagnostic accuracy of fundus photography and FFA for different degrees of diabetic retinopathy. The FFA characteristics of diabetic retinopathy was analyzed. Kappa consistency test was used to analyze the consistency between the two diagnostic methods and the clinical diagnosis results. <p>RESULTS: The sensitivity, specificity and accuracy of FFA on diabetic retinopathy were 96.8%, 97.1%, 96.9%, all of which were above the fundus photography(<i>P</i><0.05). The diagnostic accuracy of FFA in mild, moderate, severe diabetic retinopathy were 97.1%, 97.0%, 96.4%, all above the fundus photography without significance(<i>P</i>>0.05). The Kappa consistency test showed the consistency between the diagnosis results of FFA and clinical was good; and the consistency between the results of the clinical and fundus photography was only moderate. FFA test results showed that diabetic retinopathy were more visible in nasal retinal lesions and in the peripheral part from the optic disc, and intraretinal microvascular abnormalitie could be seen. In diabetic retinopathy, the number and distribution of retinal microangiomas, hemorrhagic spots, vein beads, and capillary non-perfusion zone were different. Some patients, before the formation of retinal microangiomas and hemorrhagic spots, had increased foveal thickness, focal cotton-wool spot and focal capillary non-perfusion zone. <p>CONCLUSION: FFA has high sensitivity, specificity and accuracy in the diagnosis of diabetic retinopathy. The consistency between FFA diagnosis and clinical diagnosis is strong. FFA has accurate diagnosis for suspected changes in fundus photography.

Proliferation and differentiation of endogenous neural stem cells in subventricular zone in rats after traumatic craniocerebral injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the time course of proliferation and differentiation of neural stem cells (NSCs) in the subventricular zone (SVZ) of rats following traumatic craniocerebral injury (TBI).</p><p><b>METHODS</b>Forty-eight SD rats were randomized into 3 groups, namely the control group without any treatment, the sham-operated group with scalp incision and preparation of a cranial window, and TBI group with craniocerebral injury induced by Feeney's method. With nestin and BrdU as two cell markers, NSE as the neuron-specific marker and GFAP as the glial cell marker, immunofluorescence assay with double labeled antibodies was performed to examine the proliferation and differentiation of endogenous NSCs in the SVZ at different time points after TBI.</p><p><b>RESULTS</b>s The numbers of cells positive for nestin/NSE, nestin/GFAP, BrdU/NSE, and BrdU/GFAP in the SVZ of the rats increased significantly after TBI. The positive cells began to increase at 1 day after TBI, reached the peak level at day 3 and became normal at day 14, showing significant differences between the time points of measurement following TBI and from the cell numbers in the control group measured at the same time points. The cells positive for nestin/ GFAP showed the most distinct increase in the SVZ of the rats with TBI.</p><p><b>CONCLUSION</b>TBI results in mobilization of the NSCs in the SVZ on the injured side to cause the proliferation and differentiation of the endogenous NSCs. The SVZ is one of the most important germinal centers of NSC proliferation and differentiation.</p>

Parthenogenetic activation of rabbit oocytes and effects of different additives on development of activated embryos / 中华神经医学杂志

Objective To investigate an appropriate activation method and culture system in vitro of rabbit parthenogenetic embryos to make an experimental foundation for therapeutic cloning.Methods Mature rabbit oocytes with polar body 1 (PB 1) were divided randomly into four groups and accepted,respectively,parthenogenetic activation with 1.2 kV/cm,1.6 kV/cm and 2.0 kV/cm electricity stimulation or 5 μg/mL ionomycin (ION) chemical treatment; the effects of these different treatments on activation efficiency were compared.The activated oocytes were cultured,respectively,in M199 supplemented with various concentrations of leukaemia inhibitory factor (LIF,0,10,20,40 and 80 ng/mL) or insulin+transferrin+sodium selenite (ITS,0×,1 × and 2×); and then,the rate of cleaved oocytes,percentage of morulas/blastocysts and total number of blastocysts were compared between each two groups.Results (1) Different treatments on the oocytes showed significantly different activation effects.The survival rate of oocytes activated in 5 μg/mL ION for 4 min were significantly higher than that of ones stimulated by 2.0 kV/cm electricity treatment (P=0.016).The percentages of cleaved oocytes,developing 8-16 cells and morulas/blastocysts in the ION treatment group were also significantly higher as compared with those in the 1.2 kV/cm electric treatment group (P=0.000,P=0.002 andP=0.026,respectively).(2) Significant difference in the percentage of morulas/blastocysts was noted between 20 ng/mL and 80 ng/mL of LIF supplementation (P=0.003); in addition,the total number ofblastocysts in the medium supplemented with 20 ng/mL LIF was significantly different to those with 40 ng/mL and 80 ng/mL,respectively (P=0.011,P=0.002); In experiment of ITS supplementation,significant difference in the percentage ofmorulas/blastocysts was shown between 1 × and 2× ITS supplementation (P=0.003); in addition,the total number of blastocysts in the medium supplemented with 0× or 1 × ITS (166 and 147 cells) was significantly different as compared with that with 2× ITS (78 cells,P=0.015,P=0.044).Conclusion The parthenogenesis activation of the New Zealand rabbit's oocytes treated with ION+6-DMAP shows a higher rate of cleaved oocytes and blastocysts than that with electricity stimulation; the subsequent development of parthenogenetic embryos in vitro could be significantly improved by supplementing with 20 ng/mL LIF and 1× ITS.

CatWalk gait analysis in assessment of functional changes of the hind paw after SD rats being established the artificial bladder reflex arc / 中华神经医学杂志

Objective To assess the functional changes of the hind paw after SD rats being cut offthe L5 spinal nerve by CatWalk gait analysis.Methods Eleven male SD rats were chosen; the right side of these rats,as the experimental side,was cut the ventral root and dorsal root of L5 spinal nerve,while the left side was without any operation.One day after the operation,whether the models were successfully constructed was verified by tracer experiment with wheat germ agglutinin horseradish peroxidase (WGA-HRP) in 3 rats; 1 day before the operation and 3 months after the operation,the changes of such parameters as general parameters,individual paw parameters,pain-related parameters and coordination-related parameters were assessed by using CatWalk gait analysis.Results The WGA-HRP-labeled motoneurons and fibers were not found in posterior horn and anterior horn of the experimental side of L5 spinal gray matter,but both located in the left side,indicating that the models were constructed successfully.General parameters,individual paw parameters and pain-related gait parameters could be recorded 3 months after the operation,but no statistically significant difference was noted as compared with those 1 day before the operation; however,significant decrease of values of regularity index (RI) and coupling diagonal right after-left front in rats 3 months after the operation was noted as compared with those 1 day before the operation (P＜0.05).Conclusion Only a part of coordination-related gait parameters (the lower limbs) is influenced after being cut L5 spinal nerve;therefore,L5 spinal nerve can be used for constructing of a bladder reflex arc.

Effects of hemoglobin level on Rho kinase of endothelial cells and tight junction protein claudin-5 in blood-brain barrier of rats after intracerebral hemorrhage / 中华神经医学杂志

Objective To explore the effects of hemoglobin level on Rho kinase Ⅱ of endothelial cells and tight junction protein clandin-5 in blood-brain barrier (BBB) of rats after intracerebral hemorrhage (ICH).Methods Male SD rats were randomized into normal control group (n=18) and hemoglobin inducement group (n=72); hemoglobin inducement group was divided into 4 subgroups (24 and 48 h,and 3 and 7 d after surgery,n=18).Rat ICH models in the hemoglobin inducement group were established by hemoglobin injection into the brain tissues.The expression ofRho kinase Ⅱ of endothelial cells and phosphorylated myosin light chain protein in BBB were observed by immunohistochemical staining,and BBB permeability was evaluated by Evans Blue dye; The protein and mRNA expressions of claudin-5 in the perihematomal brain tissues were detected by immunofluorescence staining and real-time quantitative PCR.Results Immunohistochemical staining indicated that expressions of Rho kinase Ⅱ of endothelial cells and phosphorylated myosin light chain in BBB of normal control group were rare,and those in the hemoglobin inducement group were high; as compared with that in the normal control group,mean optical density of Rho kinase Ⅱ and phosphorylated myosin light chain proteins in the endothelial cells of BBB in hemoglobin inducement group increased significantly at 48 h and 3 d after inducement (P＜0.05).The permeation of Evans Blue in hemoglobin inducement group significantly increased (P＜0.05).Immunofluorescence staining showed that the expression of clandin-5 in hemoglobin inducement group was uncontinuous and low as compared with that in the normal control group at 24 and 48 h and 3 d after inducement.PCR showed that the mRNA expression of claudin-5 in hemoglobin inducement group was significantly lower than that in the normal control group at 24 and 48 h and 3 and 7 d after the inducement (P＜0.05).Conclusion After intracerebral hemorrhage,hemoglobin can lead to increased expression of phosphorylated myosin light chain,and reduce the expression of tight junction protein claudin-5 of BBB by increasing the expression ofRho kinase Ⅱ in endothelial cells of BBB,which may be an important mechanism in the disruption of BBB and the occurrence of brain edema after intracerebral hemorrhage.

Affection of embolism on rat aneurysm model with new platinum coils modified by VEGF / 中华神经医学杂志

Objective To explore the embolization effect of new platinum coils coated with [4COOH-P (DLLA-co-TMC)] biodegradable polymer and released vascular endothelial growth factor (VEGF) into intracranial aneurysms on rat intracranial aneurysms. Methods A total of 54 adult healthy female SD rats were randomly divided into Group Ⅰ with general platinum coils, Group Ⅱ with polymer-coated platinum coils and Group Ⅲ with platinum coils modified with VEGF (n=18).The right common carotid arteries (CCA) of rats in each group were exposed; and the 8 mm lengths of platinum coil segments were inserted into the ligated right CCA of rats. The distal right CCA was performed ligation and restored the blood flow; 6 rats each time at 15,30 and 90 d after the surgery were chosen;and the distal right CCA was used as aneurysm models,and the left CCA without the coil placement or surgical disruption in Group I with general platinum coil was chosen as normal control.The proliferation and fibrosis of endothelial cells were observed by HE staining; von Willebrand Factor (vWF) expression was detected by immunohistochemical staining; and VEGF expression was examined by Western blotting. Results Cellular proliferation and fibrosis in Group Ⅲ with platinum coils modified with VEGF enjoyed significantly higher grade than those in Group Ⅰ with general platinum coils 10,60 and 90d after the surgery (P＜0.05); Cellular proliferation and fibrosis in Group Ⅲ with platinum coils modified with VEGF enjoyed significantly higher grades than those in Group Ⅱ with polymer-coated platinum coils 30 d after the surgery (P＜0.05).Pathological observations showed that the massive intimal hyperplasia and substantial clot completely occluded the aneurysm lumen in Group Ⅲ with platinum coils modified with VEGF; New small blood vessels having vwf-positive expression were noted in the fiberized tissues;the thrombosis in Group Ⅰ with general platinum coils and Group Ⅱ with polymer-coat platinum coils were not fully organized and showed loose hyperplasia structure with a large number of internal spaces.Western blotting indicated that the VEGF level in Group Ⅲ with platinum coils modified with VEGF were significantly higher than that in other groups 15 and 30 d after the operation,however,the VEGF level in Group Ⅲ with platinum coils modified with VEGF 90 d after the surgery was decreased because the lumen completed fibration and degradation of 4COOH-P (DLLA-co-TMC). Conclusion The VEGF-eontaining biodegradable polymer,by slowly releasing VEGF to modify the surface of platinum coils, could enhance the cellular proliferation, thrombosis and formation of dense fibrous tissue in aneurysm lumen; as compared with general platinum coils,these new platinum coils could occlude the rat aneurysm faster and more completely.

The photodynamic effects of a novel photosensitizer ZnPcS4-BSA on human U251 glioma cells in vitro / 中华神经医学杂志

Objective To report the phototoxicity effects of a novel photosensitizer ZnPcS4-BSA on photodynamic therapy (PDT) towards human U251 glioma cells in vitro. Methods The cellular uptake of ZnPcS4-BSA by U251 glioma cells was quantified by UV-spectra to determine the optimal incubation time. Human U251 glioma cells were incubated with ZnPcS4-BSA of various concentrations and received laser irradiation of different energy densities. Cell survival rates were measured by CCK-8 assay.Flow cytometer was used to detect apoptosis.Gene expressions of vascular endothelial growth factor (VEGF) were detected by Real-Time PCR in the U251 cells after PDT and β-actin was used as an internal standard. The normal U251 cells severed as controls. Results The uptake of ZnPcS4-BSA by U251 glioma cells reached the maximum after incubation for 4 hours.ZnPcS4-BSA of different concentrations without laser irradiation had no significant effects on cell survival rates (P＞0.05).Without ZnPcS4-BSA incubation,compared with 0,25,50,100,200 J/cm2 groups, the cell survival rate of the 400 J/cm2 group was significantly lower (P＜0.05), whereas no significant difference was found between any other two groups. When the U251 glioma cells incubated with 30 μ mol/L ZnPcS4-BSA for 4 hours underwent laser irradiations of 25,50,100,200 J/cm2,the cellular survival rates significantly decreased with the increased energy densities (P＜0.05). When the U251 glioma cells incubated with ZnPcS4-BSA of 20,40,60,80,100 μ mol/L for 4 hours underwent laser irradiation of 200 J/cm2, the cellular inhibition rates significantly increased with the increased concentrations (P ＜0.05). Compared with controls, the cellular apoptosis and VEGF expression significantly increased in the U251 glioma cells incubated with ZnPcS4-BSA of 20 μmol/L after laser irradiation of 100 J/cm2 (P＜0.05). Conclusion The novel ZnPcS4-BSA is a good photosensitizer for PDT towards U251 glioma cells,because the ZnPcS4-BSA-mediated PDT can induce effective apoptosis of the targeted cells.

The effect of Tenascin-R and anti-EGF-L antiserum on the cortical neurons in vitro in rats / 中华神经医学杂志

Objective To study the effect of the EGF-L functional region containing human TN-R protein in prokaryocytes and anti-EGF-L serum on the cortical neuronsin vitro in rats. Methods The anti-EGF-L serum was obtained from the rabbits immunized with the protein of EGF-L, then combine with TN-R coat petri dish to prepare various culture substrate.The effect on adhere,migrate and neurite outgrowth of neuron by differ culture substrate was observed. Results High concentration of antiserum against protein of EGF-Lwas successfully obtained from the rabbits,and it has no effect on the neuron when coated on petri dish as culture substrate with PLL in vitro,but can add the adhesive to the neuron,partly neutralize the inhibitory on the neuron neurite of TN-R.The antiserum can also allow the neuron with its neurite to migrate to the area coated with TN-R. Conclusion The anti-EGF-L serum can weaken the inhibitory on the neuron of TN-R in vitro,the function in vivo need further research,

Immunotherapy effect of dendritic cells pulsed by lysate of tumor cell Ar-He cryoablation on rat models of intracraniai gliomas / 中华神经医学杂志

Objective To investigate the role of C6 glioma cells mediated by rapid freezing and thawing ofAr-He cryoablation in the maturation of marrow-derived dendritic cells (BM-DCs) in Wistar rats,and the anti-tumor effect of these DCs on rat models of intracranial gliomas. Methods C6 glioma cells were routinely cultured in vitro; rapid freezing and thawing of Ar-He cryoablation was employed in C6 glioma cells of the experimental group, and C6 glioma cells of the negative control group were only performed insertion of the probe; blank control group (using rapid freezing and thawing of Ar-He cryoablation on the same amount of PBS) was also employed.Bone marrow-derived mononuclear cells (MNCs) were first prepared from tibia and femur bones of Wistar rats. These cells were cultured with such cytokines as recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF),recombinant interleukin-4 (rmIL-4) and tumor necrosis factor-alpha (TNFα) to induce their maturation; BM-DCs were pulsed with or without tumor cell lysate obtained by rapid freezing and thawing of Ar-Hecryoablation at a ratio of (DC:tumor cells =1:3) 7 d after that.Morphological observation of BM-DCs was performed by light microscopy and the expression of DCs costimulatory molecules CD80 and CD86 were measured by flow cytometry 48 h after the addiction; the IL-12 level in the supematant of DCs was detected by ELISA. In order to determine whether or not vaccination with C6 TP DCs can induce the therapeutic potential in the established glioma-bearing models, the C6 cells cultured in vitro were stereotaxically implanted into the left caudate nucleus of Wistar rat brain; glioma-bearing rats were injected with vaccination with DCs,cells from the blank control group and negative control group on the 3rd and 10th d. Survival time was observed and determined using the method of Kaplan-Meier and Log-Rank analysis. Results DCs from rats' bone marrow cells cultured with cytokines and pulsed with tumor lysates showed the characters of typical mature DCs.Morphologically,these cells were large with oval or irregularly shaped nuclei and with many small dendrites. Phenotypically, they expressed high levels of CD80 and CD86 antigens (71.8 1％± 1.10％ and 74.66％± 1.48％ in experimental group,49.49％±1.08％ and 51.20％±2.06％ in negative control group, and 48.47％±1.09％ and 49.53％±1.89％ in blank control group); significant difference was noted between each 2 groups (P＜0.05).Functionally,the IL-12level in the supernatant of DCs showed obvious increment in the experimental group (245.99 ±3.20pg/mL) as compared with that in the negative control group (138.68±3.20 pg/mL) and blank control group (135.16±2.88 pg/mL,P＜0.05).These cells gained the capacity of mediating immunotherapy against intracranial gliomas in rats:the median survival in the experimental group (33 d) was significantly higher than that in the negative control and blank control groups (22 and 24 d, respectively, P＜0.05).Conclusion C6 glioma cells mediated by rapid freezing and thawing of Ar-He cryoablation can induce maturation of BM-DCs in Wistar rats; these BM-DCs pulsed with tumor lysates, as new therapeutic vaccines,can mediate immunotherapy against intracranial gliomas in rats.

Quantitative CatWalk analysis of gaits in rats with 6-hydroxydopamine-induced PD / 中华神经医学杂志

Objective To investigate the probability of quantitative analysis of parkinsonian gaits in unilateral 6-hydroxyl dopamine-lesioned rats. Methods A total of 24 male Wistar rats were assigned to control group (n=6),sham-operated group (n=6) and 6-OHDA inducement group (n=12).All subjects received anesthetization and the latter 2 groups further processed with injection of equivalent volumes of normal saline and 6-OHDA into the left medial longitudinal fasciculus on the operation day,respectively. The cylinder test and Catwalk analysis were applied successively 1 week before the operation for baseline value,and 3 d, 1 and 2 weeks after the operation. Results The cylinder test indicated that the proportion of using the right forelimbs was significantly decreased as compared with that using the left forelimbs in the 6-OHDA inducement group 3 d, 1 and 2 weeks after the operation (P＜0.05), but no significant differences of using the right forelimbs were shown in the control and sham-operated counterpart (P＞0.05). Catwalk analysis showed no significant differences either on the girdle comparisons of former/hinder limbs or on pre- and post-operation comparisons of individual limbs between control and sham-operated groups (P＞0.05). In the 6-OHDA inducement group,max contact area (MCA),paw length (PL),paw width (PW) and paw area (PA) for fore girdle comparisons and MCA for hinder ones showed significant differences (P＜0.05); all the 4 parameters of right hinder limb,the PL,PW and PA of left fore limb,the PA,PL and MCA of right fore limb,the PW of left hinder limb after the operation significantly decreased as compared with those before the operation (P＜0.05). Conclusion It is applicable to detect changes in gaits of Parkinson's disease quantitatively with Catwalk analysis,especially in terms of MCA,adding new tool for further study.

Scriptaid improves early development potential of mouse somatic cell nuclear transfer embryos / 中华神经医学杂志

Objective To examine the effect of Scriptaid,a histone deacetylase inhibitor and a kind of anti-cancer drug,on development of mouse somatic cell nuclear transfer (SCNT) embryos in vitro and explore a new strategy to improve the efficiency of SCNT. Methods SCNT was carried out by pizeo-activated micromanipulator in C57/BL6 mouse,from which the oocytes were chsoen as recipients and the cumulus cells as donors.The rcconstructcd embryos were randomly divided into 5 groups and activated in calcium-free activators with 0 mmol/L (negative control group),and 50,100,250 and 500 mmol/L Scriptaid for 6 h, respectively; and then, they were transferred into KSOM medium with corresponding concentrations of Scriptaid for 4 h.The cloned embryos were finally cultured in KSOM medium for 96 h.The development (form rate) of cloned embryos and the count of blastocysts cells were recorded. Results No significant differences on the activated rate of the reconstructed embryos and the 2-cell cleavage rate were noted between each 2 groups (P＞).05).However,the form rate (24.2％) and cell numbers (56.27±2.43) of blastocysts in 250 mmol/L Scriptaid activation group were significantlyhigher as compared with those in the negative control group, and 50, 100 and 500 mmol/L Scriptaid activation groups (form rates:5.3％,6.5％,9.4％ and 6.9％; cell numbers:44.67±1.53,50.25±1.26,52.33±2.50 and 50.75±1.50,respectively,P＜0.05). Conclusion The early development potential of mouse SCNT embryos in vitro can be dramatically improved by 250mmol/L Scriptaid.

Proliferation capacity and viability of adipose-derived stromal cells labeled with ultrasmall superparamagnetic particles of iron oxide / 中华神经医学杂志

Objective To label adipose-derived stromal cells (ADSCs) with different concentrations of ultrasmall superparamagnetic particles of iron oxide (USPIO) to investigate the biological characteristics of ADSCs treated with these USPIO,and determine the optimal concentration of USPIO in labeling ADSCs in vitro. Methods USPIO with different concentrations were prepared,and the particles were endocytosed by ADSCs generated from rat adipose tissue. Eight groups (negative control group,blank control group,and 11.25,22.5,45,90,135 and 180 μg/mL treatment groups) were chosen. Labeling efficiency and cellular uptake were analyzed by Prussian blue staining. Meanwhile,proliferation capacity and viability of ADSCs were evaluated by Alamar blue assay and Cell Counting kit-8. Results ADSCs could be effectively labeled with USPIO: approximately 95％ ADSCs were labeled when they were incubated with USPIO for 24 h under the concentration of USPIO was 45 μg/mL;and approximately 100％ ADSCs were labeled when the concentration of USPIO was 90 μg/mL and above. The CCK-8 and Alamar blue tests showed that USPIO of different concentrations (11.25-90 μg/mL) had little influence on cell growth viability,and no significant difference was noted between each 2 concentration groups (P＞0.05). In a word, 45-90 μg/mL USPIO were the optimal choice for transplantion of ADSCs in vivo. Conclusion ADSCs from the adipose tissue can be effectively labeled with USPIO with minimal effect on cell proliferation and viability.

A microelectrode array for chronic in vivo single-unit recording in subthalamic nucleus of freely behaving rats / 中华神经医学杂志

Objective To introduce a bundled microelectrode array of chronic in vivo single-unit recording in subthalamie nucleus (STN) of freely behaving rats. Methods STN single cell discharge was recorded by a self-designed bundled microelectrode array.Our microelectrode array design consisted of 3 parts:(1) a 29-guage stainless steel tube was served as a guide tube to facilitate the brain penetration of microwire tips,and as a ground electrode of the brain tissues as well. (2) Five 20μm platiniridium wires were used as recording electrodes and a 50.8 μm stainless steel wire with 1 mm bare tip was employed as local potential reference; all the above resembled a guiding-hand-shaped arrangement in microwire tips,and was fixed by trny plot of carbowax moreover.(3) A male connector with end screw receptors was employed to make the array connection more stable so as to minimize the movement artifacts; six normal rats were implanted with this kind of electrode arrays,and the stability of STN single-unit recording was evaluated in the following 5 weeks. Results Twenty-seven firing units were captured during operation,of which 70.4％ (19/27) survived more than 2 d,and 9 new units were acquired within 5 weeks.No significant linear correlation of peak-to-peak amplitude was noted between each 2 different recording sections (Pearson r=-0.047,P=0.655),and whilst signal-to-noise ratio was stable with significant correlation to peak-to-peak amplitude (r=0.934,P=0.000).More than half of the initial acquired STN units retained more than 3 weeks. And there were more than 75％ r values of waveform similarities large than 0.90 of the same units across different periods. Conclusion This methodology may be appreciated for STN long-term single-unit recording with stable recording quality and favorable cell retained rate.

Correlation between NAD(P)H oxidase and malignancy grade of glioma / 中华神经医学杂志

[Objective] To detect the expression of NAD(P)H oxidase (Nox) and the content of ROS in different malignant gliomas,and explore the influence of Nox and intracellular ROS levels in the survival,proliferation,and malignant phenotype of gliomas.[Methods]Thirty human glioma specimens (10 with grade Ⅰ and I1,10 with grade II and 10 with grade IV),performed resection in our hospital from August 2007 to August 2010,were collected in our study;another 10 normal brain tissues were collected as controls.Real time PCR was used to detect the mRNA expressions of Nox(1-5);ROS level was detected by flow cytometry and Nox4 protein level was detected by immunofluorescence and Western blotting.[Results] The Noxl-5 mRNA and protein levels and ROS content were significantly different between each 2 groups (P＜0.05);Nox4 mRNA expression and ROS level significantly increased following the increment of malignancy degrees (controls＜low-grade gliomas＜grade III gliomas＜ grade IV gliomas,P＜0.05);the Nox protein expression was low in controls while that in gliomas was high,and the higher malignancy of the tumors,the higher expression levels of the tumors.[Conclusion] Nox expression and ROS content have significantly positive relations with the malignancy of the tumors,which indicates that Nox expression and ROS content might paly important roles in the occurrence of glioma and malignant proliferation.

Effect of ACNU on scores of modified neurological severity scale in rat normal brain tissues / 中华神经医学杂志

[Objective] To study the effect of ACNU,a kind of antineoplastic,on scores of modified neurological severity scale (mNSS) when it with different concentrations contacted to the local cerebral tissues of rats.[Methods]Forty-five SD rats were randomly divided into 5 groups,including Group A,B,C and D (n=10) and Group E (n=5).After being anesthetized,the parietal of the rats was opened to exposure the left cerebral hemisphere,and then the gelfoam (specification as 2 mm x2 mm x2 mam) dipped with the different doses (2,1,0.5,0.25 and 0 mg/kg) of ACNU solution were respectively applied on the surface of the rat hemisphere in Group A,B,C and D and Group E.The wound was then surgically closed.The mNSS scores in every group were respectively evaluated at 6 h,and 1,3,7,10 and 14 d after the administration of ACNU.[Results] The mNSS scores showed significant difference between each 2 groups (P＜0.05);the trends ofmNSS scores were also obviously different at different times after the administration of ACNU (P＜0.05);the changes were much stable in Group D and E;at the same time point,the mNSS scores in Group A,B and C were significantly higher than those in Group D and E (P＜0.05);the mNSS scores were increased at first and then decreased following the prolongation of administration time.[Conclusion] The 0.25 mg/kg of ACNU or lower doses of that could be considered as safe doses for applying to the normal cerebral tissue ofrats.

Proliferation characteristics of adipose-derived stem cells from neonatal suckling rats and adult ones / 中华神经医学杂志

Objective To investigate the proliferative differences of adipose-derived stem cells (ADSCs) from neonatal suckling SD rats (5-d-old) and adult ones under the same culture condition.Methods ADSCs were isolated from the subcutaneous adipose tissues of neonatal suckling SD rats and adult ones,and then,type Ⅰ collagenase digestion was employed to obtain the ADSCs; the morphology of these cells was detected.The expressions of such cell surface markers as CD45,CD29 and CD90 were observed. The number of ADSCs on the 4th d of culture under the same condition and with the same planted density was compared between the neonate and adult rats. In vitro culture of the second generation of ADSCs was performed in the 96-well plates, and CCK-8 and alamar blue kit were employed to compare and quantitate the proliferative differences; optical density was observed by microplate reader. Results The ADSCs from neonatal SD rats and adult ones expressed the stem cell biomarkers: the expression of CD45 was negative, and that of CD29 was 98.04％ and 93.17％,respectively,and that of CD90 was 94.92％ and 93.3％,respectively,for neonate SD rat and adult ones.The cell counting results indicated that the number of ADSCs from neonatal rats ([8.87±0.13]×105 cells) was larger than that of adult ones ([4.51±0.36]×105 cells) after being cultured under the same condition and at the same planted density. The optical density value of ADSCs in neonatal rats was significantly higher than that in adult ones on the 6th and 7th d of culturing detected by CCK-8 kit and on the 2nd-7th d of culturing by alamar blue assay. Conclusion The proliferative ability of ADSCs from neonatal rats is greater than that of adult ones.

Hospital first aid effect on patients with traffic accident severe craniocerebral injury: a report of 1107 cases / 中华神经医学杂志

Objective To make reference for relevant departments of our country to develop acute craniocerebral trauma prevention and control measures based on the general rule of hospital first aid by changing the links as signs of life, consciousness, pupil and time for hospital emergency in patients with traffic accident severe traumatic brain injury. Methods The clinical data of 1107 patients with severe traumatic brain injury,collected from April 1,2008 to March 30,2009,were analyzed; these patients were chosen according to the severe traumatic brain injury statistical table made by Department of Neurosurgery of Zhujiang Hospital and authorized by committee of experts from 71 other hospitals.The consciousness,pupil changes,GCS scores,blood pressure, respiration,pulse index changes on admission and at time out of emergency department were analyzed and compared; according to these results,function projections was performed; development tendency of the disease and rescue effect were judged, and the next treatment measure step was determined. Results According to the hospital emergency time,most patients got to the Emergency at 10-60 min after the injury,mostly at 10-30 min after the injury (38.9％).The consciousness,pupil changes,GCS scores,blood pressure,respiration,pulse index changes of most patients gradually developed to good trends after hospital first aid; before the patients received further specific treatment, the vital signs and indicators of them were gradually stabilized. Function projections supported these results. Conclusion Hospital first aid and effective treatment,timely controlling indicators,correct diagnosis and all reasonable measures improving hospital first aid can help to improve the first aid quality of patients with traffic accident severe traumatic brain injury.

Blockage of transforming growth factor-β signaling pathway inhibits vasculogenic mimicry in gliomas / 中华神经医学杂志

Objective To study the effect of transforming growth factor-β (TGF-β) signaling pathway blockage on vasculogenic mimicry (VM) in gliomas and explore its possible mechanism.Methods Three-dimentional culture was performed on the glioma cell lines U251 and SHG44; the effects of U251 culture supematant and TGF-β on VM formation of SHG44 cells were observed; the capability of VM formation of U251 and SHG44 cells after being treated with 0 μg/mL (PBS group),15 μg/mL TGF-β neutralizing antibody (Ab15 group) and 30 μg/mL TGF-β neutralizing antibody (Ab30 group) was evaluated.ELISA was used to detect the concentrations of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in the supematant of U251 cells from the blank group,PBS group,Ab 15 group and Ab30 group and the concentrations of VEGF and PDGF in the supernatant of SHG44 cells from the blank group,TGF-β treatment group,PBS group,Ab15 group and Ab30 group.Results VM was formed in the U251 cells while not in the SHG44 cells during the three-dimentional culture; SHG44 cells could only gather into colonies of different sizes.U251 culture supernatant could induce SHG44 cells to form VM,enjoying the most obvious effect at 24-48 h of culture; TGF-β could not induce SHG44 cells to form VM.The number of U251 cells annulation in PBS group,Ab15 group and Ab30 group decreased in sequence with significant difference (P＜0.05).The number of U251 cells armulation in SHG44 cells cultured in U251 culture supematant from the PBS group,Ab15 group and Ab30 group decreased in sequence after being added TGF-β antibody with significant difference (P＜0.05).As compared with that in the blank group and PBS group,significant decrease of VEGF and PDGF concentrations in the U251 cells from Ab15 group and Ab30 group was noted (P＜0.05); as compared with that in the blank group and TGF-β treatment group,significant increase of VEGF and PDGF concentrations in the SHG44 cells from PBS group,Ab15 group and Ab30 group was noted.Conclusion Blockage of TGF-β signaling pathways inhibits VM in glioma,and it maybe probably due to the decrease of VEGF and PDGF expressions..

Overexpression of interleukin-l7 in tumor-associated macrophages is correlated with the differentiation and angiogenesis of laryngeal squamous cell carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Interleukin-l7 (IL-17), which exerts strong pro-inflammatory effects, has emerged as an important mediator in inflammation-associated cancer. The aim of this study was to clarify the relationship between IL-17 and tumor associated macrophages (TAMs), and the correlation of the microvessel density in the development of laryngeal squamous cell carcinoma (LSCC).</p><p><b>METHODS</b>Histopathological observations and immunohistochemistry staining for IL-17, CD68, and CD34 were performed on 72 specimens (32 cases of LSCC, 20 cases of adjacent tissues of carcinoma as controls, and 20 cases of chronic hypertrophic laryngitis). Double immunohistochemical staining was done to determine which cells expressed IL-17. Real-time quantitative PCR determined the mRNA expression of IL-17. ELISA was used to detect the expression of the serum level of IL-17 in the three groups.</p><p><b>RESULTS</b>The inflammation response had increased in LSCC. Overexpression of IL-17 and CD68 protein were seen in LSCC (P < 0.01). The expression of IL-17 was different between well and poorly differentiated LSCC (P < 0.01). The IL-17 expressing cells were mainly located in macrophages (CD68(+)/IL17(+)) as demonstrated by double immunohistochemical staining. IL-17 expression significantly correlated with high microvessel density (CD34(+)) in LSCC (P < 0.05). Relatively higher mRNA expression levels of IL-17 were seen in LSCC compared to the controls (P < 0.05). The serum expression of IL-17 was similar among the three groups (P > 0.05).</p><p><b>CONCLUSION</b>IL-17 was expressed by TAMs, and IL-17 may significantly correlate to the differentiation and angiogenesis in the development of LSCC.</p>