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>

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>

Survey on clinical characteristics of pediatric allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the clinical symptom, precipitating factor, associated symptom, family history and life quality of pediatric patients with allergic rhinitis, and to analyze the characteristic of clinical symptoms.</p><p><b>METHODS</b>A questionnaire survey on pediatric AR patients since June 2008 to June 2010, one hundred and forty-eight pediatric AR patients were divided into 2 groups, group A (n = 43) included children aged from 3.2 to 6.0, group B (n = 105) included children aged from 6.1 to 14.8. The severity degree of clinical symptom was assessed by visual analogue scale.</p><p><b>RESULTS</b>Preschool age children had more severe rhinocleisis, more severe cough and less rhinorrhea than school age children (χ(2) value were 29.194, 12.277 and 16.904, respectively, P < 0.05). According to the classification criteria of ARIA 2008, preschool children had more mild intermittent AR and less moderate-severe persistent AR than school age children (χ(2) value were 20.370 and 24.546, P < 0.05). The precipitating factor of common cold, fitment, climate, environment factors were 22.3% (33/148), 5.4% (8/148), 16.2% (24/148), 3.4% (5/148), the others was 4.7% (7/148), no obvious precipitating factor was 48.0% (71/148). The rate of parent or parents who had allergic disease history was 11.5% (17/148). Quality of sleep that 66.2% (98/148) were upset and 62.2% (92/148) had no cathexis.</p><p><b>CONCLUSIONS</b>The preschool children have different clinical symptom characteristic from the school age children, and we got some clinical data of pediatric AR patients, those were beneficial to the diagnose and therapy of pediatric AR. The clinical data obtained in this study from pediatric AR patients are beneficial to the diagnosis and therapy of pediatric AR.</p>

Construction of a mouse pDsRed2-N1-SDF-1alpha eukaryotic expression vector and its expression in mouse bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector pDsRed2-N1-SDF-1alpha and observe its expression in the mouse bone marrow mesenchymal stem cells.</p><p><b>METHOD</b>SDF-1alpha gene sequence with XhoI, EcoRI restriction enzyme cutting site was amplified from the total RNA of mouse smooth muscle cells by reverse transcription-polymerase chain reaction (RT-PCR) and inserted into the eukaryotic expression vector pDsRed2-N1 encoding red fluorescent protein gene, and the insertion was verified by endonuclease digestion and DNA sequencing. Mouse bone marrow mesenchymal stem cells identified with immunofluorescence assay for vimentin expression were transfected with the constructed plasmid pDsRed2-N1-SDF-1alpha, and the expression of sdf-1alpha was detected using immunofluorescence assay.</p><p><b>RESULTS</b>The DNA fragment amplified by PCR from the total RNA was identical to SDF-1alpha from the gene library, and an identical DNA fragment was also amplified from the recombinants. Sequence analysis confirmed the successful insertion of SDF-1alpha into the pDsRed2-N1 vector and the eukaryotic expression vector pDsRed2-N1-SDF-1alpha was successfully constructed. The cultured mouse bone marrow mesenchymal stem cells positive for vimentin protein showed SDF-1alpha expression 24 h after transfection with the recombinant vector.</p><p><b>CONCLUSION</b>The pDsRed2-N1-SDF-1alpha eukaryotic expression vector constructed is capable of expression of SDF-1alpha fusion protein in the mouse bone marrow mesenchymal stem cells.</p>

Intranasal pretreatment with toll like receptor 9 ligand CpG oligodeoxynucleotides prevents the development of allergic rhinitis in juvenile guinea pigs / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of intranasal oligodeoxynucleotides with CpG motifs (CpG ODN) in prevention of allergic rhinitis in juvenile guinea pigs.</p><p><b>METHODS</b>Juvenile guinea pigs aged from 7 to 10 weeks were administrated with CpG ODN alone or combined with OVA at single dose concentration intranasally (on day 0, 5, 10, 15 in sequence) while control and blank group were administrated with saline. Both experimental and control animals were again sensitized by OVA (on day 18, 25), and 14 days after second sensitization animals were challenged by OVA intranasally (on days 39 and 46). Two hours after challenge, the animals were sacrificed. Then Hemotoxin and Eosin stain were carried out to analyze local eosinophilic reactions and nasal lesions. Local and systemic cytokines interleukin IL-5 and IFN-γ levels were examined by ELISA. Immunofluorescence was carried out with ICAM-1 antibody. Statistical analysis was performed using a SPSS 11.0 software.</p><p><b>RESULTS</b>In CpG ODN-administration or CpG ODN with OVA-administration group allergic rhinitis symptoms were not as severe as model control group (P < 0.05). Compared with the model control group, CpG ODN-administration did not increase production of OVA-specific Th1 cytokine IFN-γ but decreased productions of ovalbumin-specific Th2 cytokines IL-5 both in serum and nasal specimen (q value were 3.890 and 4.019, P < 0.05). Moreover, nasal lesions with infiltration of mean (x ± s) eosinophils (20.0 ± 9.6) in CpG group animal were prominently reduced by the CpG ODN-treatment compared with the control animals (53.5 ± 19.8) and CpG+OVA group (9.5 ± 5.7) were lower than CpG-M+OVA group (49.2 ± 18.9), the differences were significant (q value were 3.785 and 4.576, P < 0.05). Immunofluorescence results showed lower ICAM-1 expression in nasal specimen of CpG group compared with model group and CpG plus OVA group animal to CpG mimics plus OVA group (Z value were 3.697 and 3.765, P < 0.05).</p><p><b>CONCLUSIONS</b>Intranasal administration of CpG oligodeoxynucleotides with or without allergen may be an effective way to prevent the development of allergic rhinitis.</p>

Clinical characteristics in patients with non-allergic rhinitis and allergic rhinitis: preliminary analysis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To compare and analyze the clinical characteristics in patients with hyperreactive non-allergic rhinitis (HNAR) and allergic rhinitis (AR).</p><p><b>METHODS</b>A questionnaire survey on AR and HNAR patients between January and August 2009 was conducted. The clinical data of 298 AR patients and 100 HNAR patients were analyzed, including gender, age distribution, seasonal, clinical symptom and induced factors.</p><p><b>RESULTS</b>The number of male patients was more than female in AR, while in NAR, the number of female patients was more than male (χ(2) = 6.415, P = 0.01). The highest morbidity age in AR was teenagers, aged between 10 - 19 (χ(2) = 12.772, P = 0.00), while in HNAR, the highest morbidity age was middle-aged and youth, aged between 30 - 39 (χ(2) = 51.533, P = 0.00). The main onset seasons in AR was autumn, while there was no seasonal diversity in HNAR. The main allergen in AR was mugwort and ragweed, consistent with the vegetative cover characteristic in Jilin province. The main classification of AR was moderate-severe persistent (χ(2) = 123.991, P = 0.00), while the main classification of HNAR was moderate-severe intermittent (χ(2) = 97.420, P = 0.00). The clinical symptoms were significantly different between AR and HNAR except rhinocnesmus (all P < 0.05). There was consistency about non-specificity induced factors in AR and HNAR (all P > 0.05).</p><p><b>CONCLUSIONS</b>There were significant differences between AR and HNAR in sex, age, classification and seasons. The severity of clinical symptoms in AR was higher than that in HNAR except sneezing and gasping. There was consistency about induced factors in AR and HNAR.</p>

Endoscopic surgery for 89 cases of nasal inverted papilloma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of endoscopic surgery for nasal inverted papilloma.</p><p><b>METHODS</b>The clinical data of 89 patients treated with endoscopic surgery in our department from May 2003 to May 2008 were retrospectively analysed. The clinical classification was assessed by preoperative endoscopy and CT or MRI. All cases were routinely followed up from 1 to 5 years.</p><p><b>RESULTS</b>The recurrence rates in group of endoscopic management and group of endoscopic plus Caldwell-Luc management were 12.5% and 11.8% respectively (chi(2) = 0.007, P > 0.05). The recurrence rates in group of grade I, grade II and group of grade III patients were 9.1% (5/55), 17.2% (5/29) and 20% (1/5) respectively (P > 0.05). The recurrence rate in the two times operation group (27.3%) was significantly higher than that in the one time group (7.5%, chi(2) = 4.311, P < 0.05). There were 70 lesions certified to be resected completely, six of these patients (8.6%) recurred. Nineteen lesions were certified to be resected uncompletely, five of these patients (26.3%) recurred. The difference was no significant (chi(2) = 2.860, P = 0.09).</p><p><b>CONCLUSIONS</b>The endoscopic surgery in nasal inverted papillomas was an effective method with minimally invasiveness, but it still had a high recurrence. The pathological examine of surgical margin could partially judge the prognosis.</p>

Correlation analysis of two serum specific IgE test systems and skin prick test in allergic rhinitis patients / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the correlation between two serum specific IgE and skin prick test (SPT) for the diagnosis of allergic rhinitis.</p><p><b>METHODS</b>Two hundred and sixteen patients were referred to the allergist for a suspected allergic rhinitis between June and October in 2009. Patients were classified as positive for inhalant allergy if they had a positive clinical history and a related positive SPT for the suspected inhalant allergen. Statistical analysis was performed using SPSS13.0 software.</p><p><b>RESULTS</b>One hundred and fifty-eight patients had a positive SPT, comparing with the SPT, the diagnostic indexes (accuracy, sensitivity, specificity) of the ImmunoCAP system and the AllergyScreen system were 0.810 and 0.819, 0.872 and 0.780, 0.741 and 0.862 respectively. The accuracy was similar between the two systems (χ(2) = 0.112, P > 0.05). The ImmunoCAP system had a higher sensitivity (χ(2) = 7.361, P < 0.05). The AllergyScreen system had a higher specificity (χ(2) = 10.222, P < 0.05).</p><p><b>CONCLUSIONS</b>This data supported the use of ImmunoCAP system and AllergyScreen system to identify potentially significant individual allergens in the diagnosis of allergic rhinitis. The ImmunoCAP system had a higher sensitivity. The AllergyScreen system had a higher specificity. The AllergyScreen system can be used as a complementary with the ImmunoCAP system.</p>

Biocompatibility of a novel cavernous nickel-titanium alloy with rat bone marrow stromal cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the biocompatibility of a novel cavernous nickel-titanium alloy with rat bone marrow stromal cells (BMSCs) in vitro.</p><p><b>METHODS</b>Rat BMSCs were cultured on the surface of compact, microporous and macroporous nickel-titanium alloys, and the cell proliferation on day 3 during the culture was assessed using MTT assay. On day 7 of the cell culture, the cells were labeled with Hoechst33342 for cell counting under a fluorescence microscope. Scanning electron microscopy (SEM) was performed on day 7 of cell culture to observe the morphological changes of the cells.</p><p><b>RESULTS</b>The cell proliferation rate and cell numbers differed significantly between the cavernous alloy groups and the compact alloy group (P<0.05), but similar between the former two groups (P>0.05). SEM showed that compared with the compact alloy, microporous and macroporous nickel-titanium alloys had better biocompatibility with the BMSCs, and the cells on the surface of the cavernous alloys had normal cell morphology.</p><p><b>CONCLUSION</b>Cavernous nickel-titanium alloy has good biocompatibility and can promote the adhesion, aggregation and proliferation of rat BMSCs in vitro.</p>

Prokaryotic expression, purification of human LINGO-1(aa76-319) and preparation of its polyclonal antibody / 南方医科大学学报

<p><b>OBJECTIVE</b>To express and purify the fusion protein of extracellular domain of human Ig domain-containing, neurite outgrowth inhibitor (Nogo) receptor-interacting protein-1 (LINGO-1(aa76-319)) in prokaryotic cells and prepare the rabbit anti-LINGO-1 polyclonal antibody (pAb).</p><p><b>METHODS</b>The 732 bp DNA sequence of hLINGO-1(aa76-319) was obtained from pCMV-SPORT6 by PCR and inserted into pET30a(+) plasmid to construct the prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319), which was subsequently transformed into E.coli. The target fusion protein was expressed with IPTG induction and purified by Ni(2+)-NTA affinity chromatography column. The antiserum against hLINGO-1(aa76-319) was obtained from the rabbits immunized with hLINGO-1(aa76-319), and the titer of the pAb was determined using enzyme linked immunosorbent assay (ELISA) and its specificity identified using Western blotting.</p><p><b>RESULTS</b>The prokaryotic expression plasmid pET30a(+)-hLINGO-1(aa76-319) was constructed successfully. Efficient expression of the target fusion protein was achieved with IPTG induction at the optimal concentration of 0.4 mmol/L and culture temperature at 37 degrees celsius; for 2.5 h. The hLINGO-1(aa76-319) fusion protein was effectively expressed in E.coli as inclusion bodies, and the soluble protein was obtained through denaturation and refolding procedures, and the purified fusion protein showed a purity above 90%. The titer of the anti-hLINGO-1(aa76-319) pAb obtained by immunizing the rabbits with the purified protein reached 1:1.6x10(6), and Western blotting confirmed its good specificity.</p><p><b>CONCLUSION</b>The fusion protein hLINGO-1(aa76-319) with high purity has been obtained and the anti-hLINGO-1(aa76-319) pAb obtained shows a high titer and good specificity, which provide important experimental basis for further functional investigation of LINGO-1.</p>

Thymic stromal lymphopoietin expression is increased in nasal epithelial cells of patients with mugwort pollen sensitive-seasonal allergic rhinitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Excessive expression of thymic stromal lymphopoietin (TSLP) has been demonstrated in asthmatic airway epithelia and in nasal epithelia from animal models of allergic rhinitis (AR), but the evidence of expression of TSLP in nasal epithelial cells (NECs) of patients with AR is lacking. We aimed to investigate the expression of TSLP in NECs of patients with mugwort sensitive-seasonal AR and determine whether it is associated with severity of symptoms and the number of infiltrated eosinophils in nasal mucosa.</p><p><b>METHODS</b>NECs specimens were obtained by scraping with plastic curettes from the nasal inferior turbinates of patients with mugwort pollen sensitive-seasonal AR (n = 22) and nonallergic controls (n = 11) during last peak mugwort pollen season. The severity of nasal symptom was assessed using a Visual Analog Scale (VAS). In addition, serum mugwort pollen IgE levels were tested from each patient. In situ hybridization (ISH) was performed to test the messenger RNA (mRNA) of TSLP in the NECs. Furthermore, immunohistochemical staining (IHC) was scored to evaluate the expression of TSLP and eosinophil cell count was made by May-Grünwald/Giemsa staining. The correlation between expression of TSLP and all other parameters was analyzed in this study.</p><p><b>RESULTS</b>The mRNA level of TSLP was significantly increased in NECs of patients with AR compared with the nonallergic control group (P < 0.05). In addition, IHC results showed that expression of TSLP in NECs from patients with AR was up-regulated which was correlated with VAS score (r = 0.598; P < 0.05) and nasal eosinophils count (r = 0.702; P < 0.05), but it was unrelated with mugwort pollen specific IgE level.</p><p><b>CONCLUSIONS</b>These preliminary findings indicate a potential relationship between TSLP expression, severity of symptoms and nasal eosinophils count in pathogenesis of AR, but TSLP expression did not correlate with mugwort pollen specific IgE level. The elevated expression of TSLP might play a critical role in local atopical responses of AR. In the future, the TSLP has the potential to be one of the most important molecular markers for AR diagnoses and assessment.</p>

Culture and identification of neural stem cells derived from the subventricular zone of adult mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method for culturing and identifying neural stem cells (NSCs) derived from the subventricular zone (SVZ) in adult mice.</p><p><b>METHODS</b>NSCs were isolated from the SVZ of adult mouse brain and cultured in serum-free medium. Cell cloning and BrdU incorporation were performed to identify the self-renewal and proliferative capacity of the NSCs. Fluorescence immunocytochemistry was used to examine the expressions of the NSC markers nestin and SOX2, neuronal marker Tuj1, astrocyte marker GFAP and oligodendrocyte marker NG2. The expressions of nestin and SOX2 were further examined by Western blotting and RT-PCR.</p><p><b>RESULTS</b>NSCs with self-renewal and proliferative capacity were obtained from the SVZ of adult mice and grown as floating neurospheres. The NSCs expressed nestin and SOX2 and could differentiated into Tuj1-positive neurons, GFAP-positive astrocytes and NG2-positive oligodendrocytes.</p><p><b>CONCLUSION</b>This method allows simple and stable culture of NSCs from the SVZ of adult mice.</p>

Sinerem labeling and MRI tracking of neural stem cells in vivo and in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To label rat neural stem cells (NSCs) with the complex of Sinerem, the ultrasmall superparamagnetic iron oxide (USPIO), and poly-L-lysine (PLL), and evaluate the feasibility of tracking the labeled cells with magnetic resonance imaging (MRI) in vitro and in vivo.</p><p><b>METHODS</b>Sinerem was incubated with PLL to obtain the complex of Sinerem-PLL. The mesenchymal stem cells (MSCs) isolated from the bone marrow of SD rats were cultured and induced to differentiate into the neural stem cells. The second-passage cells were cultured overnight with the Sinerem-PLL complex, after which Prussian blue staining and transmission electron microscopy were performed to observe the nanoparticles in the cytoplasm. Cell apoptosis assay was performed to assess the cell viability 1 day, 1 week, and 2 weeks after the labeling. Cell tracking with 4.7 MR system was carried out in vivo and in vitro using T(2)WI and T(2)*WI sequences.</p><p><b>RESULTS</b>The NSCs could be effectively labeled with Sinerem-PLL complex with the labeling efficiency exceeding 95%. Prussian blue staining showed numerous blue iron particles in the cytoplasm, and under transmission electron microscope, these particles accumulated in the endosomes/lysosomes. The labeling did not significantly affect the cell viability and proliferation. Remarkable low signal density changes of the labeled cells was seen on T(2)WI and T(2)*WI in vivo and in vitro.</p><p><b>CONCLUSION</b>NSCs can be effectively labeled with Sinerem-PLL complex, and MRI can be used to track the labeled cells in vivo and in vitro.</p>

Time course of calpain activity changes in rat neurons following fluid percussion injury and the interventional effect of mild hypothermia / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the time course of calpain activity changes in rat neurons following fluid percussion injury (FPI) under normothermia (37 degrees celsius;) and mild hypothermia (32-/+0.5) degrees celsius;.</p><p><b>METHODS</b>In vitro cultured rat neurons were subjected to FPI followed by application of mild hypothermia for intervention at different time points, and the changes in intraneuronal calpain activity following FPI and the interventional effect of mild hypothermia on calpain activity were evaluated by UV-spectrophotometry at different time points.</p><p><b>RESULTS</b>Remarkable changes occurred in calpain activity in the neurons following FPI at 37 degrees celsius;, and mild hypothermia produced obvious interventional effect on calpain activity in close relation to the timing of intervention initiation.</p><p><b>CONCLUSION</b>Intraneuronal calpain activity changes following FPI are involved in the pathological process of cellular injury, and mild hypothermia might offer protection against traumatic brain injury to some extent by regulating calpain activity. The interventional effect of mild hypothermia is associated with the timing of the intervention initiation.</p>

Effect of superparamagnetic iron oxide labeling on neural stem cell survival and proliferation / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of superparamgnetic iron oxides (ferumoxides) on the survival and proliferation of neural stem cells (NSCs) and determine the optimal ferumoxides concentration for labeling.</p><p><b>METHODS</b>Bone marrow stromal cells (BMSCs) were obtained from rat femoral marrow and cultured in vitro to induce their differentiation into NSCs. Ferumoxides labeling of the NSCs was performed with different final concentrations of ferumoxides, and the labeling efficiency and viability of the labeled NSCs were evaluated by Prussian blue staining, MTT assay, flow cytometry and transmission electron microscope.</p><p><b>RESULTS</b>The NSCs could be effectively labeled with ferumoxides with a labeling efficiency of around 90%. Prussian blue staining showed numerous fine granules with blue staining in the cytoplasm of the labeled NSCs, and the intensity of the blue staining was in positive correlation with the ferumoxide concentration for labeling. Transmission electron microscopy of the labeled NSCs revealed the presence of numerous vesicles spreading in the cytoplasm and filled with electron-dense magnetic iron particles. The ferumoxides vesicles increased with the labeling concentration of ferumoxides, and at the final concentration exceeding 25 microg/ml, ferumoxides vesicles in the NSCs gave rise to conglomeration which hampered observation of the cellular ultrastructure by transmission electron microscope. The results of flow cytometry and MTT assay demonstrated that the cell viability, proliferation, differentiation and apoptosis of the labeled cells were affected by ferumoxides at the concentration above 25 microg/ml, but such effects could be minimal at lower concentrations.</p><p><b>CONCLUSION</b>Ferumoxides might be feasible for in vitro labeling of the NSCs with the optimal concentration of 25 microg/ml.</p>

P-JAK2 and P-STAT3 protein expression and cell apoptosis following focal cerebral ischemia-reperfusion injury in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in phosphorylated JAK2 and STAT3 protein expression of and cell apoptosis following focal cerebral ischemia-reperfusion injury in rats.</p><p><b>METHODS</b>A rat models of focal cerebral ischemia-reperfusion injury was established by middle cerebral artery occlusion using modified filament method. Immunohistochemistry and Western blot analysis were used to detect the expression of P-JAK2 and P-STAT3 proteins, and TUNEL assay was employed to examine the cell apoptosis.</p><p><b>RESULTS</b>P-JAK2 and P-STAT3 protein expression increased significantly after cerebral ischemia-reperfusion injury in rats. The immunoreactivity was prominent in the peripheral of the ischemic region and reached the peak level at 24 h of reperfusion, followed by slight decrement. The apoptotic cells increased obviously after cerebral ischemia-reperfusion injury, also reaching the peak level at 24 h of reperfusion.</p><p><b>CONCLUSION</b>The expression of phosphorylated JAK2 and STAT3 may be involved in the ischemic cellular events including apoptosis. JAK2/STAT3 signaling pathway plays a role in the pathophysiological process of cerebral ischemia/reperfusion cell injury and repair.</p>

Expression of TGFbeta1 and its type I receptors ALK1 and ALK5 mRNA in brain arteriovenous malformation / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the expression of transforming growth factor beta1 (TGFbeta1) and its type I receptors activin-like kinase 1 (ALK1) and ALK5 mRNA in the development of brain arteriovenous malformation (BAVM).</p><p><b>METHODS</b>The mRNA expressions of TGFbeta1, ALK1and ALK5 were detected with semiquantitative RT-PCR in patients with BAVM.</p><p><b>RESULTS</b>The expressions of TGFbeta1 and ALK5 mRNA increased significantly in BAVM, and their relative expression quantity were 0.777-/+0.047 and 0.585-/+0.074, respectively. However, ALK1 mRNA expression declined significantlies with a relative expression of 0.173-/+0.044 in comparison with the control group (0.720-/+0.098, P<0.01).</p><p><b>CONCLUSION</b>The balance of TGFbeta1 and its type I receptors ALK1 and ALK5 mRNA expressions may play important role in the development of BAVM.</p>

Lack of telomerase activity in rabbit bone marrow stromal cells during differentiation along neural pathway / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate telomerase activity in rabbit bone marrow stromal cells (BMSCs) during their committed differentiation in vitro along neural pathway and the effect of glial cell line-derived neurotrophic factor (GDNF) on the expression of telomerase.</p><p><b>METHODS</b>BMSCs were acquired from rabbit marrow and divided into control group, GDNF (10 ng/ml) group. Cytokine.NSCs medium (prepared by our lab, Patent No. ZL02134314. 4) supplemented with 10 percent fetal bovine serum (FBS) was used to induce BMSCs differentiation along neural pathway. Fluorescent immunocytochemistry was employed to identify the expressions of Nestin, neuron-specific endase (NSE), and gial fibrillary acidic protein (GFAP). The growth curves of the cells and the status of cell cycles were analyzed, respectively. During the differentiation, telomerase activities were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).</p><p><b>RESULTS</b>BMSCs were successfully induced to differentiate along neural pathway and expressed specific markers of fetal neural epithelium, mature neuron and glial cells. Telomerase activities were undetectable in BMSCs during differentiation along neural pathway. Similar changes of cell growth curves, cell cycle status and telomerase expression were observed in the two groups.</p><p><b>CONCLUSIONS</b>Rabbit BMSCs do not display telomerase activity during differentiation along neural pathway. GDNF shows little impact on proliferation and telomerase activity of BMSCs.</p>