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Objective:To investigate the blocking effect of non-competitive N-methyl-D-aspartic acid(NMDA) receptor antagonist Memantine on glutamate abnormal signal transmission in immature white matter induced by ischemia in vitro and in vivo. Methods:The oligodendrocyte (OL) precursor oxygen glucose deprivation (OGD) cell models of 2-day-old newborn rats were prepared and divided into the normal control group, the OGD group and the Memantine group.The extracellular glutamate level of the OL precursor was measured by high performance liquid chromatography, while the concentration of intracellular calcium and the apoptosis rate of OL precursor were detected by flow cytometry.The animal models of ischemic periventricular leukomalacia (PVL) were established and divided into the sham group, the PVL group and the Memantine group.The pathological evaluation of white matter was performed under light microscope.The positive OL expression rate of myelin basic protein(MBP) was detected by immunohistoche-mistry.The myelination of white matter was evaluated under electron microscope.Results:Compared with the normal control group in vitro, the OGD group had a higher extracellular glutamate level of the OL precursor [(24.60±2.42) μmol/L vs.(9.49±1.08) μmol/L, t=9.28, P<0.01], a higher intracellular calcium concentration [(32.9±6.9)% vs.(6.9±3.5)%, t=4.41, P<0.01], a higher apoptosis rate of the OL precursor [(24.77±2.05)% vs.(6.65±1.39)%, t=15.01, P<0.01]. After treatment with Memantine, the extracellular glutamate level [(14.70±1.70) μmol/L, t=5.68, P<0.01], the intracellular calcium concentration [(23.1±2.0)%, t=6.13, P<0.01], and the apoptosis rate of the OL precursor [(11.80±2.06)%, t=5.18, P<0.01] decreased significantly.Compared with the sham group in vivo, the white matter of the PVL group showed mild or severe pathological changes, and the PVL group had a lower MBP-positive OL expression rate in the white matter [(5.94±1.37)% vs.(15.40±3.22)%, t=4.63, P<0.01]less myelin sheaths (4.00±1.00 vs.14.67±2.70, t=6.11, P<0.01) and thinner myelin sheaths [(33.83±3.21) nm vs.(79.67±6.45) nm , t=10.43, P<0.01]. After the treatment with Memantine, the number of myelin sheaths (10.30±1.50, t=6.01, P<0.01), the thickness of myelin sheaths [(57.21±4.05) nm, t=7.47, P<0.01], and the pathological changes in the white matter of newborn rats ( Z=88.479, P<0.01) all improved markedly, and the MBP positive OL expression rate in the cerebral white matter [(11.02±1.35)%, t=4.40, P<0.05] also increased significantly. Conclusions:Ischemia-induced abnormal signal transmission of glutamate in immature white matter is the important pathway leading to ischemic PVL.Memantine can effectively block the abnormal signal transmission and thus may probably provide a new approach for the effective prevention and treatment of PVL in premature infants.
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Objective:To observe the efficacy and safety of vitrectomy combined with submacular injection of tissue-type plasminogen activator (t-PA), gas filling and anti-VEGF drugs (multiple therapy) for thick submacular hemorrhage.Methods:A retrospective case study. From January 2014 to June 2018, 24 patients (24 eyes) with thick submacular hemorrhage who received multiple therapy at the Department of Ophthalmology of Peking University Third Hospital were included in the study. Among them, there were 15 males and 9 females with the average age of 69.05±8.86 years. The average submacular bleeding time was 17.15±10.30 days, the average bleeding area was 13.85±8.63 DD. Seventeen eyes showed hemorrhagic RPE detachment. The international standard visual acuity chart was used to BCVA examination, which was converted to logMAR visual acuity in statistics. The frequency domain OCT was used to measure the height of the foveal elevation. The average logMAR BCVA of the affected eye was 1.37±0.38. The average height of the macular fovea was 824.94±294.38 μm. All eyes underwent 23G or 25G vitrectomy. During the operation, 0.1-0.5 ml t-PA (10 μg/0.1 ml) was injected under the macula. The vitreous cavity was filled with 15% C 3F 8 after gas-liquid exchange in 13 eyes, and 11 eyes were filled with sterilized air. Eleven eyes were injected with anti-VEGF drugs at the end of the operation, and anti-VEGF drugs were given PRN treatment according to the activity of the lesion. The average follow-up time after treatment was 27.90±22.21 months. The absorption of bleeding under the macula, the improvement of vision, the occurrence of rebleeding and treatment-related complications were observed and recorded. The Wilcoxon rank sum test was performed to compare the BCVA and the height of foveal elevation before and after treatment. Results:One month after the treatment, the blood in the fovea of all affected eyes was cleared. At the last follow-up, the logMAR BCVA and macular foveal elevation were 0.82±0.28 and 253.88±71.75 μm, respectively. Compared with those before treatment, the difference was statistically significant ( Z=-3.727, -3.234; P<0.001, <0.001). The average intravitreal injection of anti-VEGF drugs was 1.08 times. During the operation, a tiny hole was formed in the center of the macula when t-PA was injected under the retina. Two eyes showed mild vitreous hemorrhage early after the operation. During the follow-up period, bleeding recurred in 2 eyes. Conclusions:Vitrectomy combined with submacular injection of t-PA, gas filling, and anti-VEGF drugs is an optimal combination for the treatment of thick submacular hemorrhage. It can effectively remove submacular hemorrhage, improve vision, reduce foveal elevation with good safety.
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<p><b>OBJECTIVE</b>To analyze clinical manifestations and genetic mutation in a child with severe short stature and other malformations.</p><p><b>METHODS</b>The child has undergone history taking and physical examination. Genome DNA was extracted from peripheral blood samples of the proband and her family members. Candidate genes were captured with Agilent SureSelect and sequenced on an Illumina platform. Suspected mutation was verified by Sanger sequencing.</p><p><b>RESULTS</b>The patient, a six-year-and-10-month old girl, presented with non-symmetrical short stature, dysmorphism, abnormalities of limbs and spine, amblyopia of left eye, and cataract of right eye, in addition with frequent respiratory infection and micturition. Laboratory testing suggested 25-hydroxy vitamin D deficiency (18.9 ng/mL). Spine X-ray showed multiple malformations with centrums. Her mother also featured short stature (138 cm). Her aunt had short stature (130 cm) and limb-length discrepancy. Her little brother was 2.5 years old, and his height was 81 cm (-3.4 SD). Exome sequencing revealed a heterozygous mutation c.184C to T (p.Arg62Trp) in the proband and her mother. The same mutation was not found in her father and brother.</p><p><b>CONCLUSION</b>The patient was diagnosed with X-linked chondrodysplasia punctata 2. Mutation of the EBP gene probably underlied the disease in this family.</p>
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Objective To study the protective effect of curcumin on precurosor oligodendrocytes (preOLs) damage induced by hydrogen peroxide (H2O2),and explore its mechanism.Methods (1) In vitro primary culture ofpreOLs was performed; and 0 (normal controls),100,250 and 500 μmol/L H2O2 were added for 30 min; the apoptosis and death of preOLs were observed by Hoechst33342/PI double staining.(2) The preOLs were divided into normal control group,model group,and 5,10 and 20 μmol/L curcumin treatment groups; cells the later three groups were given 5,10 and 20 μmol/L curcumin for one h,and the later four groups were given 100 μmol/L H2O2 for 30 min; MTT assay was,then,employed to detect the cell viability.(3) The preOLs were divided into normal control group,model group,and 10 μmol/L curcumin treatment group; the apoptosis ofpreOLs was detected by Annexin V/FITC flow cytometry; Western blotting was used to detect the protein expressions of B cell lymphoma/leukemia-2 (Bcl2),Bcl-2 associated X protein (Bax) and cleaved caspase-3 and caspase-9; activities of total superoxide dismutase (T-SOD),glutathione peroxidase (GPx) and catalase (CAT) and malondialdehyde (MDA) were detected by spectrophotometry.Results (1) As compared with those in the normal control group,significantly decreased number of normal healthy cells and statistically increased apoptotic and necrotic cells in the 100,250 and 500 μmol/L H2O2 treatment groups were noted (P<0.05); 100 μmol/L H2O2 treatment group had larger number of apoptotic cells than that of necrotic cells,while 250 and 500 μmol/L H2O2 treatment groups had larger number of necrotic cells than that of apoptotic cells.(2) Cells from 5,10 and 20 μmol/L curcumin treatment groups had significantly higher preOLs viability than those from model group,and 10 and 20 μmol/L curcumin treatment groups had significantly higher preOLs viability than those from 5 μmol/L curcumin treatment group (P<0.05).(3) As compared with the model group,the 10 μmol/L curcumin treatment group had obviously decreased apoptotic and necrotic cells,increased activities of T-SOD,GPx and CAT,increased GSH level and decreased MDA concentration,up-regulated Bcl-2 expression,and inhibited Bax,caspase-3 and caspase-9 expressions,with significant differences (P<0.05).Conclusion Curcumin has protective effect on preOLs against oxidative injury through effectively reducing lipid peroxidation,regulating Bcl-2/Bax expression and suppressing caspase-3 and caspase-9 activation.
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Background The pathogenesis of dry eye has not been clearly established.There are more and more evidences to suggest that it is associated with ocular surface inflammation mediated by immunity.But how T helper cell 17 (Th17) plays its role in dry eye remains unclear.Objective The aim of this study was to investigate the expressions of Th17 related cytokines in ocular surface with dry eye.Methods A prospective cohortl study was designed.This protocol was approved by Ethic Commission of Peking University Third Hospital,and written informed consent was obtained from each subject prior to entering this cohort.Twenty female patients with Sjigren syndrome (SS group),20 patients with non-SS dry eyes and 20 normal volunteers were recruited in Peking University Third Hospital during 2011-2012 duration and all the subjects were menopausal female with the age 50 years old or more.The ocular surface disease index (OSDI)questionnaire designed by Schiffman was performed firstly,and then tear breakup time (BUT),corneal fluorescein staining and Schirmer Ⅰ test (S Ⅰ t) were carried out in the subjects.Expression of Th17 related cytokines mRNA were measured using PCR-Array.The correlation between IL-17A and ocular surface parameters was analyzed.Results The OSDI scores of the SS dry eye group,non-SS dry eye group and normal control group were 50.00 (33.50,66.50),45.00 (35.50,55.00) and 3.00 (0.00,5.00),the S Ⅰ t values were 2.50 (1.00,4.00),5.00 (2.00,5.00) and 15.50 (10.00,18.50),the BUT were 2.00 (1.00,4.00),4.00 (3.00,5.00) and 10.00 (10.00,12.00),the corneal fluorescein staining score were 8.50 (6.00,12.00),5.50 (4.00,7.00)and 0.00 (0.00,0.00),respectively,and significant differences were seen among the SS group,non-SS dry eye group and normal control group (x2=34.11,28.13,93.66,92.25,all at P<0.01).The relative expression values of IL-17A mRNA,IL-6 mRNA,IL-8 mRNA,IL-22 mRNA and IL-23 mRNA in the ocular surface were 1.98±0.16,11.64±1.32,6.67±1.12,1.88±0.18 and 1.78±0.17 in the SS group patients,and 1.45±0.17,1.32±0.14,1.12 ±0.13,1.23 ±0.15 and 1.23 ±0.13 in the non-SS dry eye group patients,respectively,with significant differences between the two groups (all at P<0.01).IL-17A level on the ocular surface was significantly negative correlated with BUT (r =-0.56,P<0.01) and positive correlation with corneal fluorescein staining scores (r=0.44,P=0.01).Conclusions Expressions of Th17 related cytokines in the ocular surface increased in patients with dry eye,especially in those with SS.IL-17A level in ocular surface is associated with BUT and corneal fluorescein staining scores,suggesting that immune is involved in the pathogenesis and devlopment of dry eye.
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Objectives To explore the effect of application of uridine diphosphate-glucose (UDP-glucose) on self-repairment potentiality of immature white matter (WM) in vivo. Methods Five-day-old rats were randomly divided into sham, periventricular leukomalacia (PVL) and UDP-glucose groups. The PVL model was constructed in the PVL and UDP groups, and UDP-glucose (2000mg/kg) was induced by an intraperitoneal injection at once to the rats of UDP group. PVL in-duced proliferation and differentiation of WM-glial progenitor cells invivo were detected by using the three-label lfuorescent immunoanalysis, the apopotosis in WM cell was observed by TUNEL test, and the pathology of WM and myelination were evaluated by light and electron microscopy at day 7 and day 21 after PVL model construction. Results The numbers of new WM-progenitors (NG2+), oligodendrocytes (OLs) progenitor marker (O4+), OL precursors, cyclic nucleotide phosphodiesterase (CNPase+), immature OLs and myelin basic protein (MBP+), and mature OLs in the UDP-glucose group are signiifcantly grea-ter than those in the PVL group at each time interval after induction of PVL (P<0.05). The numbers of the apoptotic cells in UDP-glucose group are less than those in the PVL groups. Under light and electron microscopy, the white matter pathological changes and myelination were found to be better than those in the PVL group (P<0.05). Conclusion The application of UDP-glucose can induce the WM-progenitors to activate, proliferate and differentiate into immature and mature OLs. UDP-glucose can also signiifcantly reduce the apoptotic rate of the WM-new glia cells;improve the white matter pathological changes and the myelin formation.
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Objectives To explore the clinical features, diagnosis and treatment of Crohn’s disease complicated by erythromelalgia (EM) in a pediatric case. Methods The clinical manifestation, results of laboratory testing and endoscopy, mutational analysis of the SCN9A gene, and the follow-up record were collected and analyzed based on review of literature to a thirteen-year-old girl with Crohn’s disease complicated by erythromelalgia. Results The patient experienced symptoms of anorexia, fatigue, diarrhea, dark red and swelling skin, increased skin temperature and burning pain in her both lower extremities during the course of disease. The endoscopic ifndings included multiple ulcerations and polypoid protrusion lesion in colon, and the pathological examination showed the local abscess formation in colonic mucosa. The mutation in SCN9A gene of the child was excluded by gene analysis. The symptoms were gradually ameliorated after treatment using prednisone and mesalazine combined with dipyridamole and low-molecular-weight heparin calcium. Conclusions Crohn’s disease complicated by erythromelalgia is rare. The pathogenesis may relate to immune factors, thrombocytosis, and hyper-coagulable states, etc. The combination of glucocorticoids, 5-aminosalicylic acid and anticoagulants may lead to a better therapeutic effect.
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OBJECTIVE@#To investigate 19 short tandem repeat (STR) loci in Chinese Kazakh population in Barkol County with Goldeneye™20A multiplex amplification system.@*METHODS@#DNA samples were screened from 81 unrelated individuals. The 19 loci were D8S1179, D21S11, CSF1PO, D3S1358, D7S820, TH01, D13S317, D2S1338, D18S51, D16S539, TPOX, vWA, D19S433, D5S818, PentaD, PentaE, D6S1043, D12S391 and FGA. The PCR products were analyzed and genotyped by ABI3130XL sequencer.@*RESULTS@#These loci were highly polymorphic. The combined power of discrimination was 0.999999999 and the combined paternity of exclusion was 0.999998914.@*CONCLUSION@#Goldeneye™20A multiplex amplification system is very useful in forensic case investigation for Barkol Kazakh population.
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Humans , Asian People , Genetics , China , Forensic Genetics , Gene Frequency , Genetic Loci , Genetics, Population , Microsatellite Repeats , Genetics , Polymorphism, GeneticABSTRACT
Objectives To investigate the incidence of brain injuri in premature infants in ten hospitals of seven large cities in China sponsored by the Subspecialty Group of Neonatology of Pediatric Society, China Medical Association. Methods All premature infants with gestational age less than 37 weeks in ten hospitals were given routine cranial ultrasound within three days of birth, and then repeated every 3-7 days till the discharge from the hospital during January 2005 to August 2006. Results Incidence of intraventricular hemorrhage (IVH) and severe IVH were 10.8% (406/3 768) and 2.4% (92/3 768) with 22.6% (92/406) for grade 1, 54.7% (222/406) for grade 2, 17.2% (70/406) for grade 3 and 5.4% (22/406) for grade 4 in nine hospitals; incidence of periventricular leukomalacia (PVL) and cystic PVL were 2.3% (112/4 933) and 0.3% (16/4 933) with 85.7% (96/112) for grade 1, 12.5% (14/112) for grade 2, and 1.8% (2/112) for grade 3 including all ten hospitals, respectively. Risk factors associated with increased severity of IVH were vaginal delivery (OR = 1.874, 95% CI = 1.172 - 2.997, P < 0.01), perinatal asphyxia (OR = 1.598, 95% CI = 1.077 - 2.372, P < 0.05), mechanical ventilation (OR = 3.988, 95% CI= 2.448 -6.948, P< 0.01), and amniotic fluid contamination (OR = 2.192, 95% CI = 1.054 - 4.544, P< 0.05). Risk factors that might result in the development of cystic PVL were vaginal delivery (OR = 1.400, 95% CI = 1.186 - 1.652, P < 0.001) and mechanical ventilation (OR = 3.000, 95% CI = 1.015 - 8.864, P < 0.05). Conclusions These data reflect basically the prevalence of brain injuriy in premature infants in major cities of China. However, more than 60% of population lives in the rural area, further multicenter investigation including the rural area is expected to be undertaken in future.
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BACKGROUND: Periventricular leukomalacia is a major syndrome of premature infant brain injury, which has been not prevented and cured yet. Theoretically, neural stem cells which were transplanted into white matter with an absence of oligodendroglial cells might be an ideal method to cure periventricular leukomalacia. OBJECTIVE: To prepare the multi-lineage potential of neural stem cells for the use of intraventricular transplantation. METHODS: Cerebral cortex was obtained from 12-14-day fetal rats and sectioned into 1.0-mm~3 sections. The single cell suspension was separated and purified. The neurospheres were incubated with DMEM/F12 culture medium containing fetal bovine serum to observe primary and passage culture of neural stem cells. The differentiation of neural stem cells was determined using immunohistochemical method. RESULTS AND CONCLUSION: The viability of cultured neural stem cells was (94.3±2.2)%. The neurosphere was formed at day 3 after primary culture. The proliferation of neurosphere slowed down after 10-passage culture, and some cells became old. All neurospheres were positively Nestin-staining, thus they were considered as neural stem cells. A further incubation of 4-passage neurospheres, immunohistochemical method indicated that the neurosphere was positively GFAP, β-tublin, and O4 staining, respectively. This suggested that cultured neural stem cells are able to self-renew, proliferate, and differentiate into neurons, astrocytes and oligodendroglial cells.