Objective To evaluate the clinical value of 6 h 99mTc-diethyl iminodiacetic acid (99mTc-EHIDA) planar hepatobiliary scintigraphy (HBS),6 h tomographic HBS and 24 h planar HBS in diagnosis on biliary atresia(BA).Methods Seventy cases(32 male,38 female) with continuous jaundice received planar and tomographic HBS in Beijing Friendship Hospital from Jan.2005 to Dec.2007.The mean age was 48.7 d (29 d-4 months).According to final diagnosis,all cases were divided into BA group (45 cases) and non-BA group (25 cases).All cases fasted at least 4 hours before HBS.The equipment was 3 head IRIX from Philips company with low energy high resolution collimator.The tracer was 99mTc-EHIDA and the radiochemistry purity was more than 95 percent.The dosage was 7.4 MBq/kg.All diagnosis demonstrated by operation pathology and clinical follow-up.All cases received HBS at 5,10,15,20,30 min and 1,6 h after tracer injection.HBS would ended if radioactivity appeared in gallbladder or intestine.These cases would received tomographic HBS and 24 h HBS if radioactivity did not appear in gallbladder or intestine at 6 h post injection.All these images were analyzed by 2 or more nuclear medicine physicians.Results There were not radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 27 cases,which suggested the BA.There were radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 30 cases,which suggested the non-BA.Positive rate of 6 h tomographic HBS was significantly higher than that of 6 h planar HBS and there was significantly difference between the 2 methods.Positive rate of 6 h tomographic HBS was significantly higher than that of 24 h planar HBS and there was significant difference between the 2 methods.Conclusions 99mTc-EHIDA HBS is a noninvasive,safety,valuable examing method and has definitely clinical value in the diagnosis on BA.The clinical value of 6 h tomographic HBS is significantly higher than that of 6 h planar HBS and 24 h planar HBS.

Objective To study the effect of injured podocytes on glomerular maturation and its underlying mechanism in neonatal mice.Methods Single i.p.injection with puromycin aminonucleoside (PA,0.1 mg/g BW) was given to ICR neonatal mice at day 1 after birth (1 dpp). Littermates injected with normal saline (NS) were used as control.Animals were examined for urine protein,blood pressure,kidney weight/body weight (KW/BW),renal histology at 2,4,8,12, 30,60 and 90 dpp (n=6～9 for each group).Immunohistochemistry and quantitative RT-PCR were performed to examine the expression of WT-1,CD31,VEGF,Flk-1,Ang-1,Ang-2,Tie-1 and Tie-2.Results Mice with PA injection had lower kidney weight and body weight at all time points as well as lower KW/BW at 4,8,12 dpp when compared with NS controls.Electron microscopy revealed nearly complete foot process effacement and segmental microvillous transformation as early as 1 day after PA injection.PA-injected kidneys showed fewer capillary loops and decreased maturation index as well as less CD31-positive endothelium in cortical glomeruli at 12 dpp. Glomerular mesangial injury and developing glomerulosclerosis along with proteinuria were noted in PA-injected kidneys starting from 30 dpp.Significantly increased systolic blood pressure was detected at 60 dpp in PA mice.Compared with NS injection,PA injection significantly induced decreased mRNA expression of Flk-1 and Tie-2 as well as increased expression of Ang-1,without obvious changes of VEGF at 2 dpp.Conclusions Podocytes in neonatal kidney of ICR mice are susceptible to PA. Such podocyte injury can alter the expression of VEGF and angiopoietin system in glomeruli,leading to abnormal development of glomerular capillaries,and subsequent proteinuria,hypertension and glomerulosclerosis.

BACKGROUNDCellular cardiomyoplasty by transplantation of various cell types has been investigated as potential treatments for the improvement of cardiac function after myocardial injury. A major barrier for the clinical application of cell transplantation is obtaining sufficiently large quantities of suitable cells. Allogeneic cellular cardiomyoplasty may provide an alternative source of abundant, transplantable, myogenic cells by in vitro manipulation of cardiac fibroblasts using chemicals including 5-azacytidine. This study evaluated cardiomyogenic differentiation of cardiac fibroblasts, their survival in myocardial scar tissue, and the effect of the implanted cells on heart function.

METHODSPrimary cardiac fibroblasts from neonatal rats were treated with 5-azacytidine (10 µmol/L) or control. Treatment of 5-azacytidine caused myogenic differentiation of cultured cardiac fibroblasts, as defined by elongation and fusion into multinucleated myotubes with sarcomeric structures as identified by electron microscopy, and positive immunostaining for cardiac specific proteins, troponin I and β-myosin heavy chain (β-MHC) and the gap junction protein connexin 43. The myogenic cells (1.0 × 10⁶) were transplanted into the infarcted myocardium 2 weeks after coronary artery occlusion.

RESULTSBy 1 month after transplantation, the converted fibroblasts gave rise to a cluster of cardiac-like muscle cells that in the hearts occupied a large part of the scar with positive immunostaining for the myogenic proteins troponin I and β-MHC. Engrafted cells also expressed the gap junction protein connexin 43 in a disorganized manner. There was no positive staining in the control hearts treated with injections of culture medium. Heart function was evaluated at 6 weeks after myocardial injury with echocardiographic and hemodynamic measurements. Improvement in cardiac function was seen in the hearts transplanted with the 5-azacytidine-treated cardiac fibroblasts which was absent in the hearts treated with control.

CONCLUSIONThe 5-azacytidine has a unique capacity to induce myogenesis in cardiac fibroblasts in vitro and transplantation of cardiac-like muscle cells into ventricular scar tissue improves myocardial function.

Animals ; Azacitidine ; therapeutic use ; Cells, Cultured ; Fibroblasts ; drug effects ; transplantation ; ultrastructure ; Immunohistochemistry ; Microscopy, Electron, Transmission ; Myocardial Infarction ; drug therapy ; therapy ; Rats

Objective To compare the pharmacognosy characteristics of aerial roots of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem.. Methods Fresh aerial roots were harvested and were used as the experimental samples. Stereoscopy was used for the observation of macroscopic appearance of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem.,and the microscope was used for the examination of their microscopic features of the velamen surface, cross section of root tip, cross section and longitudinal section of the posterior root, and powder. Results The appearance characteristics of the two species were as follows:the number of aerial roots of Ficus microcarpa Linn. f. was more,and the diameter was smaller than that of Ficus elastica Roxb. ex Hornem. The root tips of Ficus microcarpa Linn. f. aerial roots were light yellow turning to yellow-white, covered with gray or yellowish-white lenticels;the root tips of Ficus elastica Roxb. ex Hornem. aerial roots were light yellow or yellow, covered with gray lenticels. Microscopic identification results of the two plants were as follows:the primary xylems of transverse section of root tips and posterior roots of Ficus microcarpa Linn.f. and Ficus elastica Roxb. ex Hornem. were different,the former being five to seven heptarch,and the latter being six to eleven heptarch. Both of the two species had non-articulated unbranched laticifers in their longitudinal section of posterior root, and the diameter of Ficus. elastica Roxb. ex Hornem. was slightly larger than that of Ficus microcarpa Linn. f.. The powder of Ficus microcarpa Linn. f. was red brown,with spiral and pitted vessels;Ficus elastica Roxb. ex Hornem. was yellow brown,with single small and large pitted vessels,and the color of its fiber was shallow or nearly colorless or even transparent, with lines of cluster crystal. Conclusion The results will provide evidence for the identification , exploitation and utilization of Ficus microcarpa Linn . f . and Ficus elastica Roxb. ex Hornem.

Objective To obtain the elasticity value of solid breast lesions with supersonic shear wave elastrography (SWE) and apply the binary Logistic regression in order to evaluate the value of SWE in differential diagnosis of benign and malignant breast lesions. Methods SWE quantitative elastography was preformed in 91 breast lesions of 91 patients in Zhenghai Longsai Hospital to obtain the maximum and mean elasticity value (Emax, Emean). And receiver operating characteristic (ROC) curves were used to assess the diagnostic performance. A Logistic regression for the gray scale ultrasound and the elastic modulus was conducted with multiple variables including Emax, Emean, border, echo, form, calcification. Results Pathological examination showed 73 benign lesions and 18 malignant lesions. Emax and Emean of malignant lesions were obviously higher than those of benign lesions [(99.73±41.15) kPa vs (38.59±14.28) kPa, (61.45±24.88) kPa vs (23.46±11.44) kPa, t=-15.05,-14.12, both P=0.000]. The area under the ROC curve of Emax and Emean were 0.932 and 0.915. Taking 63.70 kPa as the threshold of Emax, the sensitivity was 77.8%and the speciifcity was 97.3%. Then taking 44.22 kPa as the threshold of Emean, the sensitivity was 83.3%and the speciifcity was 94.5%. The results of Logistic regression analysis showed:the 3 most effective variables were Emax, border of the lesions and Emean. Conclusions The multivariate analysis model of binary Logistic regression can select the valuable indexes of differential diagnosis of benign and malignant breast lesions. SWE plays an important role in differentiating benign and malignant lesions and it is valuable in clinical practice.

Idiopathic pulmonary fibrosis (IPF) is characterized by myofibroblast foci in lung parenchyma.Myofibroblasts are thought to originate from epithelial-to-mesenchymal transition (EMT).Wnt1 and lithium chloride (LiCl) induce EMT in alveolar epithelial cells (AECs),but the mechanisms are unclear.AECs were treated with Wnt1 and LiCl,respectively;morphological change and molecular changes of EMT,including E-cadherin,fibronectin,and vimentin,were observed.SB203580 was administrated to test the role of p38 MAPK signaling in EMT.Then AECs were treated with siRNAs targeting p38 MAPK to further test the effects of p38 MAPK,and the role was further confirmed by re-expression of p38 MAPK.At last β-catenin siRNA was used to test the role of β-catenin in the EMT process and relationship of β-catenin and p38 MAPK was concluded.Exposure of AECs to Wnt1 and LiCl resulted in upregulation of vimentin and fibronectin with subsequent downregulation of E-cadherin.Wnt1 and LiCl stimulated the p38 MAPK signaling pathways.Perturbing the p38 MAPK pathway either by SB203580 or through p38 MAPK siRNA blocked EMT and inhibited fibronetin synthesis,which were reversed by transfection of p38 MAPK expression plasmid.β-catenin siRNA attenuated the EMT process and decreased p38 MAPK phosphorylation,indicating that β-catenin is involved in the EMT-related changes through regulation of p38 MAPK phosphorylation.These findings suggest that p38 MAPK participates in the pathogenesis of EMT through Wnt pathway and that p38 MAPK may be a novel target for IPF therapy.

The eye tumors are divided into intraocular and extraocular, and mainly involve lids, conjunctiva, cornea, and orbit. The authors analysed histopathological and clinical review of 9 cases of eye tumors that had experienced.
Conjunctiva ; Cornea ; Orbit

Adoptive immunotherapy using allogeneic natural killer (NK) cells provides to be useful in recipients after allogeneic hematopoietic stem cell transplantation (Allo-HSCT), but its application has been limited by the inability to obtain sufficient numbers of pure NK cells. This study was aimed to optimize the expansion of high purity NK cells from human peripheral blood. First, the NK cells were isolated from PBMNC by using miniMACS (magnetic cell-selection) and NK Cell Isolation Kit II. Then the isolated cells were cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of IL-2 and/or IL-12, IL-15 for 15 days. Cultures were fed with fresh media and cytokines every 3 days, and were evaluated for cell expansion, phenotype, and cytotoxicity at the end of the culture period. The results showed that in group IL2 + IL15 and IL2 + IL15 + IL12, cells were expanded 50.46 +/- 4.31 and 52.35 +/- 6.72-fold respectively, much higher than others (P<0.01), but no significant difference between themselves (P>0.05). And the purity of CD3(-)CD56(+) NK cells was over 94% in all groups except the control. The cytotoxicity of expanded NK cells cultured with cytokines was significantly higher than the starting population at different E:T ratio (P<0.01), although the cytotoxicity of IL2 + IL15 + IL12 group was slightly higher than that of IL2 + IL15 group, but no significant difference between themselves (P>0.05). It is concluded that high purity of NK cells can be efficiently expanded in culture with IL2 + IL15.
CD3 Complex ; analysis ; CD56 Antigen ; analysis ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Cell Separation ; Cells, Cultured ; Culture Media ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Killer Cells, Natural ; cytology ; immunology ; Leukocytes, Mononuclear ; cytology

OBJECTIVETo study the predictors of depression in patients with chronic prostatitis (CP).

METHODSWe enlisted 49 CP patients in this study, and evaluated their clinical symptoms with NIH-CPSI and IIEF-5, their depression symptoms with the Self-Rating Depression Scale (SDS) and Symptom Checklist 90 (SCL-90), their trait-oriented coping styles with the Trait-Oriented Coping Styles Questionnaire (TCSQ), and their illness perceptions with the Illness Perception Questionnaire (IPQ-R). We conducted Pearson correlation analysis on the correlation of depression with the clinical symptoms of the patients and used multivariate Logistic regression models to analyze the predictors of their depression.

RESULTSPain or discomfort, urination symptoms, impact of symptoms and total score in NIH showed a significant positive correlation with depression symptoms, but a negative correlation with erectile function (r = 0.32, 0.31, 0.35, 0.38, and -0.36, P<0.05). Besides, 43.4% of the depression symptoms in the CP patients could be explained by negative trait-oriented coping and disease impact factors (R2 = 0.434, adjusted R2 = 0.456, F = 14.853, P<0.001).

CONCLUSIONDepression symptoms are closely associated with clinical symptoms, and negative trait-oriented coping and disease impact factors are the main predictors of depression in CP patients.

Adaptation, Psychological ; Adult ; Chronic Disease ; Depression ; etiology ; Humans ; Logistic Models ; Male ; Multivariate Analysis ; Prostatitis ; psychology ; Self-Assessment ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo study the effect of gamma-interferon (IFNgamma), tumor necrosis factor related apoptosis inducing ligand (TRAIL), and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms.

METHODSThe expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFN-gamma, TRAIL, IFNgamma + TRAIL, IFN-gamma + Caspase 8 inhibitor + TRAIL, IFNgamma + cisplatin + TRAIL, and IFNgamma + etoposide + TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay.

RESULTSCaspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNgamma. SH-SY5Y cells themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNgamma were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFN-gamma + TRAIL group was significantly higher than those of control group, IFN-gamma group, TRAIL group, and inhibitor group (P < 0.01). There was no significant difference among IFN-gamma + TRAIL group, IFNgamma + cisplatin + TRAIL group, and IFNgamma + etoposide + TRAIL group.

CONCLUSIONSIFNgamma could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 8 ; genetics ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Etoposide ; pharmacology ; Humans ; Interferon-gamma ; pharmacology ; Neuroblastoma ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology