Aim To investigate the effects of miR-122a on blood-spinal cord barrier after spinal cord ischemia-reperfusion injury in rats.Methods Thirty-six SD rats were randomly divided into three groups:group of sham(S group),group of control(C group)and group of miR-122a antagomir(M group).Rats in S group were subjected to exposure of aorta arch but without occlusion.Spinal ischemia-reperfusion injury was induced by clamping the aorta arch for 14 min in C group and M group.Rats in M group and C group were intrathecally injected with miR-122a antagomir or antagomir control daily for three times after injury.The miR-122a expression in injured spinal cord tissue was detected by real-time PCR.The occludin expression in injured spinal cord tissue was detected by Western blot.The permeability of blood-spinal cord barrier was examined using evans blue as a vascular tracer.The neurological motor function was evaluated by Basso Beattie Bresnahan score.Results Compared with S group,the expression of miR-122a was increased,the expression of occludin was decreased,the permeability of blood-spinal cord barrier was increased,and neurological motor function score was decreased significantly in C group(P<0.05).Compared with C group,the expression of miR-122a was decreased,the expression of occludin was increased,the permeability of blood-spinal cord barrier was decreased,and neurological motor function score was increased significantly in M group(P<0.05).Conclusion miR-122a can regulate the expression of occludin and change the permeability of blood-spinal cord barrier.

Humans ; Integrative Medicine ; Medicine, Chinese Traditional

A strain producing eggshell membrane protease (ESM protease) was isolated from the soil and identified as Pseudomonas aeruginosa. The enzyme isolated from the fermentation liquid of this strain and purified by ammonium sulfate precipitation, quadratic anion-exchange chromatography exhibited eggshell membrane degrading activity of 304.5 U/mg. By SDS-PAGE, the protein molecular mass is 32 kD. The N-terminal amino acid sequence of this protease is: Ala, Glu, Ala, Gly, Gly, Val, Ala, Gly, Lys, Glu, Asp, Ala, Ala, Glu, Leu.

Objective To investigate the effects of intrathecal transplantation of bone marrow stromal cells (BMSCs) on inter cellular cell adhesion molecule-1 (ICAM-1) expression and blood spinal cord barrier following spinal cord ischemia reperfusion injury.Methods Ninety Sprague Dawley rats were randomly divided into three groups:sham (Sham group),ischemia-reperfusion injury (I/ R group),and BMSCs transplantation (BMSCs group).Spinal I/R injury was induced by clamping the aortic arch between left common carotid artery and left subclavian artery for 14 min in I/R group and BMSCs group.Sham group was subjected to exposure of aortic arch but without occlusion.I/R group and BMSCs group were intrathecally injected with phosphate buffered saline (PBS) or BMSCs (2 × 106) two days before injury.At 1 d,3 d,and 7 d after injury,neurological function was evaluated and damaged lumbosacral seg ment was removed for measurement of blood spinal cord barrier permeability and ICAM-1 protein expression.Results Compared with Sham group,neurological function score was significantly lower:1 d (F =38:59,P =0.001),3 d (F =31.34,P =0.001),and 7 d (F =27.71,P =0.001) ; ICAM-1 expression was increased 1 d (F =34.33,P =0.001),3 d (F =29.76,P =0.001),and 7 d (F =23.65,P =0.001),and blood spinal cord barrier permeability was higher:1 d (F =42.57,P =0.001),3 d (F =32.75,P =0.001),and 7 d (F =26.89,P =0.001) in I/R group.Compared with I/R group,neurological function score was increased:1 d (F =16.62,P =0.001),3 d (F =21.54,P =0.001),and 7 d (F =12.84,P =0.002) ; ICAM-1 expression was decreased:1 d (F =19.84,P =0.018),3 d (F =17.38,P =0.008),and 7 d (F =22.46,P =0.007),and blood spinal cord barrier permeability was lower:1 d (F =22.38,P =0.016),3 d (F =27.59,P =0.009),and 7 d (F =23.25,P =0.001) in BMSCs group.Conclusions Intrathecal transplantation of BMSCs inhibited ICAM-1 expression and decreased blood spinal cord barrier permeability,and then attenuated spinal cord ischemia-reperfusion injury.

AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.

Objective The aim of this study is to detect the mRNA and protein expression levels of ERCC1 and XPF genes among different age groups of healthy Chinese Han individuals,and to analyze the correlation between the mRNA and protein expression levels andthe age of individuals in order to find new molecular markers for forensic age estimation.Methods Peripheral blood samples were obtained from 150 unrelated healthy Chinese Han individuals.The plasma was centrifuged from the whole blood by gradient centrifugation,and the totalRNA was extractedwithTrizol fromperipheral blood mononuclear cells(PBMCs).Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to quantitatively analyze the mRNA relative expression levels of ERCC1and XPF in PBMCs.Enzyme linked immunosorbent assay (ELISA) was used to quantitatively analyze the protein expression levels of ERCC1and XPF in plasma.Results There were no significant differences in the mRNA relative expression levels of ERCC1 and XPF in PBMCs between males and females(P＞0.05).Significant differences were found in the mRNA relative expression levels of ERCC1 and XPF between different age groups (P＜0.05).Regression analysis showed thatthe mRNA relative expression levels of ERCC1 and XPF were both negatively correlated with age.The correlation coefficients(r) were-0.578 and-0.844,respectively.When the age was used as independent variable(x) and the mRNA expression relative level as dependent variable (y),the fitting curveswere Y=3.3E-5X2-0.0261X+1.9175 (R2=0.3244,P＜0.01),Y=0.0003X2-0.0459X+2.0439 R2=0.729,P＜0.01),respectively.There were no significant differences inthe protein expression levels of ERCC1 and XPF in plasma between different age groups or genders (P＞0.05).Conclusion The mRNA relative expression levels of ERCC1 and XPF in PBMCsdeclined with the increase of age,however,the protein expression levels in plasma were unrelated to age.ERCC 1 and XPF genes can be used asnew molecular markers for forensic age estimation,so as to providetheoretical basis for establishing the mathematical model of ERCC1/XPF genesin concern ofindividual ages.

Purpose To investigate the clinicopathologic features, diagnosis and differential diagnosis of ovarian juvenile granulosa cell tumor (JGCT).Methods The history records, pathologic features and immunophenotype of 8 cases of JGCT were retrospectively evaluated and their prognosis was achieved by follow-up.Results The age of patients ranged from 6～21 years old,with an average age of 15.1 years.The main clinical manifestations included an abdominal mass, ascites and isosexual pseudoprecocity. Cut surface of the tumor was typically solid with cysts formed. The histopathological changes displayed solid nests, diffuse sheet, multiple round or ovoid follicles in variable size.Macrofollicles could be seen in some cases.The follicular pattern consisted of small cystic cavities containing eosinophilic secretions. The tumor cells were round or polygonal, medium in size. The tumor cells had abundant pale or slightly eosinophilic cytoplasm, round nuclei with fine chromatin. Nuclear grooves were inconspicuous.Mitosis figures could be found. Immunohistochemical results showed that the tumor cells expressed inhibin-α,CD99,vimentin; while Melan-A,calretinin and S-100 were positive staining in part of the cases.CKpan,EMA,PLAP,Syn and CgA were negative in all the cases.Conclusions Ovarian juvenile granulosa cell tumor is a rather rare, low malignant tumor with good prognosis. Its diagnosis depends on the histologic and immunohistochemical findings and clinical features. Its differential diagnosis includes adult granulose cell tumor, hypercalcaemic type small cell carcinoma, carcinoid and dysgerminoma.

Objective To evaluate the association between vitamin D deficiency and diabetic nephropathy in type 2 diabetic patients.Methods A total of 594 patients with type 2 diabetes were enrolled from the inpatients of the Nanjing Medical University Affiliated Nanjing Hospital.Fasting serum lipid profile,25-hydroxycalciferol vitamin D and urinary albumin excretion rate were investigated.The relationship between nephropathy and vitamin D deficiency (＜ 20 μg/L) or insufficiency (20-＜ 30 μg/L) was analyzed.Results Nephropathy was found in 177 subjects (29.8％) with albuminuria in 141 and proteinuria in 36 subjects.Vitamin D deficiency was found in 180 subjects and insufficiency in 157 subjects.The proportion of vitamin D deficiency was higher in the individuals with nephropathy than those without nephropathy (36.2％ vs 27.8％,P ＜0.05).The urinary albumin excretion rate was significantly higher in the patients with vitamin D deficiency than those with normal vitamin D concentration [(123.0 ± 299.2)mg/24h vs (47.6 ±97.1) mg/24h,P ＜0.01].The prevalence of nephropathy was higher in the patientswith vitamin D deficiency than those with normal vitamin D concentration (35.6％ vs 26.1％,P ＜ 0.05),while the prevalence of proteinuria was higher in patients with vitamin D deficency (12.2％ vs 3.1％,P ＜0.01).Logistic regression analysis demonstrated that vitamin D deficiency was associated with nephropathy (OR 1.57,95％ CI 1.04-2.37),even after the adjustment for age,gender,hypertension,dyslipidemia,smoking status,use of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers (OR 1.78,95％ CI 1.12-2.81).The Vitamin D concentration was significantly negatively correlated with urinaryalbumin excretion rate (r =-1.783,P ＜ 0.001).Conclusions Type 2 diabetic patients have a high prevalence of vitamin D deficiency.Vitamin D deficiency is independently associated with diabetic nephropathy.

Adult ; Exons ; Female ; Gastrointestinal Stromal Tumors ; genetics ; metabolism ; pathology ; Gene Deletion ; Humans ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism

OBJECTIVETo investigate the role of p38 MAPK on ischemic postconditioning (IPO) attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI).

METHODSForty adult male SD rats were randomly divided into 5 groups based upon the intervention (n = 8): control group (C), LIR group (I/R), LIR + IPO group (IPO), IPO + solution control group (D), IPO + SB203580 group (SB). Left lung tissue was isolated after the 2 hours of reperfusion, the ratio of wet lung weight to dry lung weight (W/D), and total lung water content (TLW) were measured. The histological structure of the left lung was observed under light and electron transmission microscopes, and scored by alveolar damage index of quantitative assessment (IQA). Apoptosis index (AI) of lung tissue was determined by terminal deoxynuleotidyl transferase mediated dUTP nick end and labeling (TUNEL) method. The mRNA expression and protein levels of and Bax were measured by RT-PCR and quantitative immunohistochemistry (IHC).

RESULTSCompared with C group, W/D, TLW, IQA, AI and the expression of Bax of I/R were significantly increased, the expression of Bcl-2 and Bcl-2/Bax were significantly decreased (P < 0.05, P < 0.01), and was obviously morphological abnormality in lung tissue. Compared with I/R group, all the indexes of IPO except for the expression of Bcl-2 and Bcl-2/ Bax were obviously reduced, the expression of Bcl-2 and Bcl-2/Bax were increased (P < 0.05, P < 0.01). All the indexes between D and IPO were little or not significant( P > 0.05). The expression of Bcl-2 and Bcl-2/Bax of SB were significantly increased and other indexes were reduced than those of IPO (P < 0.05, P < 0.01).

CONCLUSIONIPO may attenuate pneumocyte apoptosis in LIRI by inactivation of p38 MAPK, up-regulating expression of Bcl-2/Bax ratio.

Alveolar Epithelial Cells ; cytology ; Animals ; Apoptosis ; Disease Models, Animal ; Ischemic Postconditioning ; Lung ; blood supply ; enzymology ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology ; pathology ; prevention & control ; bcl-2-Associated X Protein ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism