Objective To analyse the characteristics of orbital benign and malignant masses on diffusion weighted imaging in combination with conventional MR imaging and evaluate the diagnostic value of apparent diffusion coefficient in distinguishing benign and malignant orbital lesions.Methods Seventy- seven cases with orbital masses,including fifty-five benign lesions and twenty-two malignant tumors,who underwent conventional MRI and diffusion imaging scanning were studied with use of a 1.5 T magnetic resonance system.Quantitative ADC measurements of masses(ADC_M)and of the white matter of eontralateral temporal lobe(ADC_W)were made with two different b-values of 0 and 1000 s/mm~2.The ADC ratio(ADCR)of the lesion to the control was calculated.The receiver operating characteristic curves(ROC) were constructed using various cut points of ADC_M and ADCR for different parameters to differentiate between benign and malignant masses.The area under the ROC curve for each parameter was also calculated. Results All cases were proved by histopathology.The mean ADC_M and ADCR of benign orbital masses were(1.56?0.75)?10~(-3)mm~2/s and 1.85?0.91,respectively.The mean ADC_M and ADCR of malignant orbital masses were(1.09?0.42)?10~(-3)mm~2/s and 1.28?0.53,respectively.There were significant difference both between ADC_M and ADCR of benign and malignant masses(t=2.803,2.735,P

OBJECTIVETo establish the method for cotransferring human A20 gene and human heme oxygenase-1 (HO-1) gene into the isolated rat islets using lentiviral transfection system, and to study the protective effect of A20 and HO-1 protein against the apoptosis induced by cycloheximide (CHX) and TNF-α, and finally to explore the underlying mechanism.

METHODThe A20 gene and HO-1 gene were cloned and inserted into the lentiviral transfection system. The efficacy of gene transfer was measured by the intensity of the enhanced green fluorescent protein (EGFP) fluorescence-positive islets. Western blot was applied to verify the expression of the A20 and HO-1 genes. To induce apoptosis in vitro, the isolated islets were treated with CHX+TNF-α, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and the fluorescence-activated cell sorting (FACS) methods were used to evaluate the apoptosis of the islet cells and Western blot was used to detect caspase-3 activation.

RESULT(1) A20 and HO-1 genes were introduced into the isolated islets by lentiviral transfection, both of the genes were highly expressed in the islets after 96 hours culture detected by Western blot method. (2) The insulin levels in the cell culture medium from A20 and/or HO-1 transgenic islets were significantly higher than that in non-transgenic controls (P < 0.01). (3)After CHX + TNF-alpha treatment, the cell culture medium insulin concentration in the A20 gene transfected group [(93.58 ± 4.12)µg/ml], HO-1 gene transfected group [(88.98 ± 4.77) µg/ml ] and A20/HO-1 co-transfected group [(103.33 ± 3.16) µg/ml] were significantly higher than that in the EGFP group [(9.03 ± 0.65) µg/ml ] and the control group [(8.86 ± 0.38) µg/ml] (P < 0.001). Minimum expression level of the activated caspase-3 was found in the A20/HO-1 co-transfected group.

CONCLUSIONThe lentiviral gene transfer system was an efficient and stable gene transfer vector, the over-expressed A20 and HO-1 protein delivered via lentivirus could preserve rats' islets function and act against the apoptosis induced by CHX and TNF-α.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Flow Cytometry ; Genetic Vectors ; Heme Oxygenase-1 ; genetics ; metabolism ; Humans ; Insulin ; metabolism ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Islets of Langerhans ; drug effects ; enzymology ; physiology ; Lentivirus ; genetics ; Male ; Nuclear Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection ; methods ; Tumor Necrosis Factor alpha-Induced Protein 3 ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the mechanism of Ca(2+) on the apoptosis induced by hyperthermia in neonate rat hippocampal neurons to provide the applicative evidence of dantrolene for preventing brain injuries.

METHODSDantrolene, Ca(2+) specific blocking agent, was used in the hyperthermia-induced apoptosis of primary hippocampal neurons in vitro to observe its effect on the apoptosis, fluorescent intensity, and dynamic change of Ca(2+) by flowcytometry and laser confocal microscopy.

RESULTSThe rate of apoptosis was decreased significantly after hyperthermia treatment by dantrolene sodium. The intracellular Ca(2+) fluorescent intensity in 42 degrees C treatment group (107.35 +/- 6.0) was significantly lower than that in control group (159.12 +/- 33.8). The concentration of Ca(2+) began to decrease 20 approximately 25 s after adding dantrolene sodium, and reached the lowest level about 50 s later, and then kept lower than the basal level.

CONCLUSIONDantrolene sodium has an important protective effect on hippocampal neurons apoptosis induced by hyperthermia and may have some applicative value of preventing heat-induced brain injury.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Dantrolene ; pharmacology ; Female ; Hippocampus ; cytology ; drug effects ; Male ; Neurons ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Temperature

Through systemically sorting and studying literature of Chinese medicine, this article pointed out that leech used by the traditional Chinese medicine in ancient time has the features of small, living in water, able to suck blood of animal and people. The species of leeches having these features were Hirudo nipponia Whitman, H. pulchra Song, Poecilobdella nanjingensis sp. Nov. , P. manillensis (Lesson) and P. hubeiensis Yang, which were not fully coincidence with the species recorded in Chinese Pharmacopeia of 2010 edition. We suggests that species of leech in Chinese Pharmacopeia be revised: H. nipponica Whitman should be kept, P. manillensis (Lesson) should be added in, Whitmania pigra Whitman and W. acranulata Whitma should be temporarily reserved, and H. pulchra Song, P. nanjingensis sp. Nov. , and P. hubeiensis Yang should be considered.
Animals ; Conservation of Natural Resources ; statistics & numerical data ; Humans ; Leeches ; classification ; Medicine in Literature ; Medicine, Chinese Traditional

OBJECTIVETo investigate the effect of PD98059 on the proliferation and cell cycle of rat hepatic stellate cells (HSCs) stimulated by acetaldehyde and explore its mechanism.

METHODSRat HSCs stimulated by acetaldehyde were incubated with different concentrations of PD98059. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. The mRNA of cyclin D1 and CDK4 were examined by RT-PCR.

RESULTS20, 50, 100 micromol/L PD98059 could significantly inhibit the proliferation of HSCs stimulated by acetaldehyde in a does-dependent manner (0.109+/-0.020, 0.081+/-0.010 and 0.056+/-0.020 vs 0.146+/-0.030, F=31.385, P<0.05) and provoke G0/G1 phase arrest of HSCs stimulated by acetaldehyde in a does-dependent manner (61.9%+/-6.3%, 64.1%+/-3.3% and 70.9%+/-4.8% vs 55.2%+/-4.4%, F=16.402, P<0.05). 50, 100 micromol/L PD98059 could markedly inhibit cyclin D1 mRNA expression of HSC stimulated by acetaldehyde (0.56+/-0.04 and 0.46+/-0.03 vs 0.65+/-0.07, F=68.758, P<0.05) and CDK4 mRNA expression (0.39+/-0.07 and 0.33+/-0.05 vs 0.50+/-0.06, F=29.406, P<0.05).

CONCLUSIONThe Erk signal transduction pathway plays an important role in regulating the proliferation and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde, which may be partly related to its regulative effect on the expression of cyclin D1 gene and CDK4 gene

Acetaldehyde ; pharmacology ; Animals ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Hepatocytes ; drug effects ; Proto-Oncogene Proteins ; Rats

OBJECTIVETo develop a simple, rapid and reliable method of purifying Sprague-Dawley (SD) rat islets by sequential filtration through two cell strainers of different sizes and to evaluate the efficacy of the method.

METHODSIslets were isolated from 8 to 12-week-old clean grade Sprague-Dawley rat pancreases using the standard collagenase digestion procedure and purified with either the generally used Ficoll density gradient method or the innovative two-step sequential filtration method. The purity and vitality of the isolated islets were visualized and assessed with DTZ and AO/PI staining. Glucose stimulating tests were performed to assay cell activity, and immunohistochemical staining was used to evaluate the synthesis function of islet cells.

RESULTSThe yield of islets in the two-step filtration method group was 782±115 IEQ per rat, which was significantly higher than in the conventional Ficoll density gradient method group (598 ± 135 IEQ per rat, P < 0.01). Purity of the isolated islets in the two-step filtration method group was 90%-100% and vitality was over 95%. In the conventional Ficoll density gradient method group, islet purity was 65%-85% and vitality was 85%-95%. With regard to the high-sugar stimulation test in the two-step filtration method group, insulin concentrations in islets cultured for 24 hours were significantly higher than in those that were freshly purified (76.9 ± 6.1 μg/L vs 49.4 ± 3.9 μg/L; P < 0.01).

CONCLUSIONSA two-step sequential filtration method for rat islet purification was developed and the method was simple and reliable, with high islet vitality, purity and yield.

Animals ; Cell Separation ; methods ; Female ; Filtration ; Immunohistochemistry ; Insulin ; biosynthesis ; Islets of Langerhans ; cytology ; Male ; Rats ; Rats, Sprague-Dawley

AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P ＜ 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P ＜ 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.

Objective To explore a sensitive method and index to evaluate renal functional reserve (RFR) in patients with early diabetic nephropathy (DN) using protein loading-scintirenography.Methods Fifty subjects were studied and divided into 3 groups.Group one (G1) consisted of 14 healthy volunteers;Group two (G2) consisted of 15 patients with type 2 diabetes mellitus (DM) and normoalbuminuria; Group three (G3) consisted of 21 patients with type 2 DM and microalbuminuria.All subjects underwent baseline and protein loading-99 Tcm-DTPA scintirenography within one week.RFR was calculated as the difference between stimulated and baseline glomerular filtration rate (GFR), time of peak ( Tb ), time of half excretion ( C1/2 ), residual rate at 20 min ( C20/b ) .Variance analysis and t-test were used to analyze the group differences.Results ( 1 ) The RFR in terms of GFR had statistical difference between any two groups (t=14.884, 32.180, 16.042, all P＜0.01).After protein-loading, the GFR of G1, G2 and G3 increased 20.1, 10.9 and 2.2 ml·min-1·1.73 m-2 respectively.Therefore, the RFR decreased before microalbuminuria in type 2 DM patients.(2)There was statistical difference between the RFR of G1 and G2 in terms of C1/2 (t = 5.505, P＜0.05 ), and between G1 and G3 ( t = 8.914, P＜0.01 ).(3) There was statistical difference of the RFR in terms of TP between G1 and G3 (t = 5.690, P ＜ 0.01 ).(4) There was statistical difference of the RFR in terms of C20/b between G1 and G3 (t= 4.376, P＜0.05 ).Conclusions 99Tcm-DTPA protein loading-scintirenography is an effective method for measuring RFR to evaluate early DN in type 2 DM patients.

Objective To evaluate the effect of PTH gene polymorphisms on bone mineral density (BMD) at multiple skeletal sites in normal females.Methods PTH gene phenotype was determined by PCR-RFLP of restriction enzyme Bst BⅠin 596 females aged (46.3?13.7) years (20-80 years),and PCR products with or without enzymolytic site were considered as genotype B or genotype b respectively.BMDs of the anteropesterior spine (AP) and supine lateral spine (Lat) of lumbar vertebrae (L_1-L_4),femoral neck (FN),total hip (T-hip), Ward's triangle (Ward),Trochanter (Troch),forearm [radius+ulna ultradistal (RUUD) and total area of radius + ulna (RUT) ] were measured by DEXA (QDR4500A).Results (1) Hardy-Weinberg equilibrium was evident for PTH polymorphisms.The frequencies of genotype were BB 0.784,Bb 0.208,bb 0.008 and frequencies of alleles B,b were 0.888 and 0.112 respectively in 596 normal females.Frequencies of BB,Bb,bb genotypes were 0.781,0.210,and 0.009 respectively in 347 postmenopausal women and their frequencies of alleles B,b were 0.886,0.114.No significant difference was found between post- and premenopausal women in genotype frequen- cy.(2) Comparing their BMDs of AP,Lat,FN,T-hip,Ward,Troch,RUUD and RUT,there was no significant difference between BB and Bb genotypes of pre- and postmenopansal women groups.(3) Logistic regression analysis failed to show any statistical difference between normal and osteoporosis women with regard to PTH phenotype.Conclusion PTH gene polymorphism has little effect on BMD in normal females.

Objective To evaluate MR imaging findings of uveal metastases.Methods MR imaging findings of 20 cases with uveal metastases comfirmed by pathology or follow-up were retrospectively analyzed.MR imaging was performed in 20 patients,of which postcontrast T1-weighted imaging was performed in 19 patients including dynamic contrast enhancement scanning in four cases.Results Metastatic tumor was found in the iris and ciliary body in two cases,and in choroid in 18 cases.The tumor demonstrated slightly hypointense signal on Tl-weighted imaging and isointense signal on T2-weighted imaging in two cases,isointense signal on T1-weighted imaging and isointense signal on T2-weighted imaging in nine cases,isointense signal on T_1-weighted imaging and slightly hyperintense signal on T_2-weighted imaging in three cases,isointense signal on T_1-weighted imaging and slightly hypointense signal on T_2- weighted imaging in three cases,slighdy hyperintense signal on T_1-weighted imaging and slightly hypointense signal on T_2-weighted imaging in two cases,and slightly hyperintense signal on T_1-weighted imaging and slightly hyperintense signal on T_2-weighted imaging in one case.The tumor appeared as mild thickness of the wall of the globe in eight cases,a crescent mass in three cases,a fusiform mass in seven cases,and a nodule in two cases.Nineteen patients showed moderate or marked enhancement on postcontrast T_1-weighted imaging.The time-intensity curve of dynamic contrast enhancement in four patients suggested a rapid enhancement and slow washout pattern.Retinal detachment was observed in 11 patients and abnormal signal intensity within the vitreous body was seen in two cases.Conclusion MRI can display the location,shape, signal characteristics,and enhancement pattern of uveal metastases,contributing to diagnosis and differential diagnosis.