Objective To investigate the effect of brain-derived neurotrophic factor (BDNF) gene modified neural stem cells (NSCs)transplantation on cerebral ischemic injury in rats.Methods NSCs from newborn rat hippocampus were isolated,cultured in a medium containing fibroblast factor (FGF) in vitro. Their proliferation and differentiation were detected by immunohistochemistry.Virus vectors pLXSN-BDNF were built and BDNF were transfected into NSCs.Biological activity were detected to obtained engineering stem cells of BDNF protein with secretary activity.Middle cerebral artery occlusion (MCAO) models were made and transplanted with NSCs-BDNF by stereotaxic technique.The effect of transplantation on MCAO models was evaluated histologically and behaviorally.Results Statistical analysis showed that the behavioral performance of the animals improved after transplantation (Longa mark being 1.343?0.293 four weeks after stem cell transplantation).The number of hippocampal dentate gyrus neurons increased to 87.5%?6.6% , four weeks after stem cell transplantation on Nissle stained hippocampal sections,which was significantly different from that of controls.Positively BrdU stained neural stem cells revealed by immunohistochemistry in the cultured cells and in the transplanted brain sections,indicated that the engineering cells transplanted had survived in the host brain and began to proliferate.Conclusion Transplantation of BDNF genetically modified NSCs can effectively promote the recovery from cerebral ischemic injury.

Objective T o prom ote the further research on body stature estim ation and the innovative ap-plications based on the distances betw een the anatom ical landm arks on body torso surface. Methods A specification for the collection of distances betw een the anatom ical landm arks on body torso surface w as established. T he data of 933 cases of adult population in Y angtze R iver D elta region w ere collected. M ultiple linear regression m ethod w as used to statistical analyse and establish the regression equation of stature estim ation. Results A regression equation about 5 variables including gender (x1), cervical verte-brae-coccyx line (x2), sterna-pubis line (x3), distance betw een acrom ion and iliospinale anterius (x4) and shoulder breadth (x5), and stature (y) w as established, y=105.406+5.414 x1+0.436 x2+0.286 x3+0.225 x4+0.193 x5. Conclusion T he m ethod is suitable for the rapid, sim ple and accurate estim ation of stature for the forensic experts.

OBJECTIVETo explore the effect of Sedum sarmentosum Bunge Extract (SSBE) on severe acute pancreatitis (SAP) induced acute lung injury (ALI) model rats and their excessive inflammatory reactions.

METHODSForty-two healthy adult male Sprague-Dawley (SD) rats were randomly divided into 3 groups, the sham-operated control group (C), the SAP group (SAP), and the SSBE treated group (SSBE), 14 in each group. SAP induced ALl rat model was induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the pancreatic duct. SSBE (100 m/kg) was administrated subcutaneously after the establishment of the SAP model. Equal dose of SSBE was injected again 12 h later. Equal volume of normal saline was administrated in the same way for rats in the C group and the SAP group. Rats were sacrificed after successful modeling and samples taken at 12 and 24 h. Pathological changes in the pancreas and the lung tissue were observed under light microscope. The ascites, serum amylase (AMS), wet/dry proportion (W/D) of the lung tissue, activities of myeloperoxidase (MPO), interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were also measured.

RESULTSAscites and serum AMS activities significantly increased; MPO, IL-1, IL-6, TNF-alpha contents, and W/D ratio also significantly increased in the SAP group, when compared with the C group (P<0.05). Compared with the SAP group, those parameters were all attenuated in the SSBE group at 12 and 24 h (P<0.05, P<0.01). Pathological changes in the pancreas and the lung tissue were alleviated in the SSBE group under light microscope. The injury degree ranged between that of the C group and the SAP group.

CONCLUSIONSSBE could relieve the ALl in SAP model rats, which could be achieved through alleviating inflammation responses of SAP rats.

Acute Lung Injury ; drug therapy ; etiology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Interleukin-1 ; Interleukin-6 ; Lung ; Male ; Pancreas ; Pancreatitis ; complications ; drug therapy ; Peroxidase ; Rats ; Rats, Sprague-Dawley ; Sedum ; Taurocholic Acid ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate the clinical features of Epstein-Barr virus-related hemophagocytic lymphohistiocytosis (EBV-HLH), to analysis the outcome of HLH-2004 protocol, and to explore the prognostic factors in EBV-HLH patients.

METHODSThe clinical features at onset and outcome of HLH-2004 protocol from 83 pediatric patients with EBV-HLH enrolled from January 2006 to December 2009 in our hospital were analyzed retrospectively. Univariate and multivariate COX regression analysis were used to identify statistically significant prognostic factors.

RESULTS(1) Among the 83 patients, 45 were males and 38 were females. The age of onset ranged from 6 months to 14 years 4 months. 44 patients were treated with HLH-2004, and 3-year overall survival (OS) was (55.8 ± 7.9)%. (2) The most common clinical features of EBV-HLH included high fever, cytopenia, hepatosplenomegaly, and coagulopathy; The respiratory symptoms, angina phlogistic, skin rashes, neurologic abnormality were rare. 97.3% of patients showed an elevation of serum ferritin, liver dysfunction and lipid metabolism disorders was found in most of EBV-HLH patients. 89.0% of patient had hemophagocytosis in bone marrow at diagnosis of EBV-HLH. (3) COX regression analysis revealed that anemia degree, serum albumin < 30 g/L, CD4:CD8 abnormity, NK cell < 3%, treatment protocol were related with the prognosis significantly (P < 0.05).

CONCLUSIONEBV-HLH in pediatric patients has severe clinical feature and poor prognosis. HLH-2004 protocol is an effective treatment for patients with EBV-HLH. Symptomatic treatment can't rescue the patients of EBV-HLH.

Adolescent ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; drug therapy ; Female ; Herpesvirus 4, Human ; Humans ; Infant ; Killer Cells, Natural ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; virology ; Male ; Prognosis ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo isolate and culture the murine fetal epidermal stem cells (ESCs) and folliculus pili cells (FPCs) in vitro, and to observe the regeneration of hair follicle and epidermis after cografting of ESCs and FPCs.

METHODSThe ESCs were isolated by adhering to murine type IV collagen and were cultured in conditional medium. The expression level of beta1-integrin and keratin 15 in ESCs was detected. At the same time, the cell cycle and clony forming eficiency (CFE) in ESCs were also determined. The FPCs were isolated and cultured and inoculated in fibrin-gel to form FPCs-gel. A full skin equivalent was prepared by combining ESCs with FPCs-gel and was grafted onto total skin loss wounds on the back of BALB/C nude mice. The histological changes of the wounds and the hair follicles were observed at 8 - 10 weeks after the grafting.

RESULTSThere were high level expressions of beta1-integrin and keratin 15 in murine fetal ESCs. It was indicated by cell cycle analysis that cells in G1 stage accounted for 94.9% of the cells, while that in S stage, 3.5%, suggesting slow cell cycle. Nevertheless, the keratinocytes in G1 stage accounted for 74.1% and that in S stage, 17.5% of cells in control group. The CFE of ESCs was 15.3%, and it was much higher than that in control group (6.7%). The newly formed hair follicles could be found in the grafted rats but not in the control group 8 - 10 weeks after the wound healing in nude rats.

CONCLUSIONThe ESCs could be successfully isolated and cultured in vitro and might participate in the formation of hair follicle structure under the induction of FPCs.

Animals ; Cells, Cultured ; Dermatologic Surgical Procedures ; Epidermis ; Fetus ; Hair Follicle ; cytology ; physiology ; Immunohistochemistry ; Integrin beta1 ; analysis ; Keratin-15 ; Keratins ; analysis ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Regeneration ; Skin ; injuries ; Skin Transplantation ; methods ; Skin, Artificial ; Stem Cells ; chemistry ; cytology ; Wound Healing

Fluorescence intraoperative cholangiography (IOC) is a potential alternative for identifying anatomical variation and preventing iatrogenic bile duct injuries by using the near-infrared probe indocyanine green (ICG).However,the dynamic process and mechanism of fluorescenceIOC have not been elucidated in previous publications.Herein,the optical properties of the complex of ICG and bile,dynamic fluorescence cholangiography and iatrogenic bile duct injuries were investigated.The emission spectrum of ICG in bile peaked at 844 nm and ICG had higher tissue penetration.Extrahepatic bile ducts could fluoresce 2 min after intravenous injection,and the fluorescence intensity reached a peak at 8 min.Inaddition,biliary dynamics were observed owing to ICG excretion from the bile ducts into the duodenum.Quantitative analysis indicated that ICG-guided fluorescence IOC possessed a high signal to noise ratio compared to the surrounding peripheral tissue and the portal vein.Fluorescence IOC was based on rapid uptake of circulating ICG in plasma by hepatic cells,excretion of ICG into the bile and then its interaction with protein molecules in the bile.Moreover,fluorescence IOC was sensitive to detect bile duct ligation and acute bile duct perforation using ICG in rat models.All of the results indicated that fluorescence IOC using ICG is a valid alternative for the cholangiography of extrahepatic bile ducts and has potential for measurement of biliary dynamics.

Objective To measure the concentration of intracellular free Ca2+ ([Ca2+]i) in neuron-like cells resulted from rat bone mesenchymal stem cell (BMSCs) differentiation induced by salvia miltiorrhiza injection and provide some theoretical basis for the BMSCs transplantation. Methods The rat BMSCs were separated from rat bone marrow and cultured in vitro. After induced by basic fibroblast growth factor and 10mL/L salvia miltiorrhiza injection, the cells were identified with immunofluorescence staining against NeuN. The same procedure was performed on primarily cultured hippocampal neurons. Then, the [Ca2+]i of the differentiated neuron-like cells was determined and compared with primarily cultured hippocampal neurons. Results The BMSCs after induced by basic fibroblast growth factor and salvia miltiorrhiza injection expressed neuronal phenotypes similar to the cell appearance of neurons with NeuN. The average fluorescence intensity of the neuron-like cells derived from BMSCs was 984.75±79.51, while the average fluorescence intensity of the primarily cultured hippocampal neurons was 769.42±60.93. No significant difference was found between them (P>0.05). Conclusion The neuron-like cells from rat BMSCs differentiation induced by salvia miltiorrhiza injection possess certain neuronal properties.

Objective To investigate the clinical application value of Loewenstein Occupational Therapy Cognitive Assessment (LOTCA) battery in Chinese patients with post-stroke aphasia.
Methods Cognitive functions of 59 Chinese patients with aphasia following a stroke were assessed with the Chinese version of the second edition of LOTCA battery and their linguistic functions were tested with the Western Aphasia Battery (WAB) Scale, respectively. The results of LOTCA were analyzed and compared across different groups, in the light of gender, age, educational background, the length of illness, and the degree of aphasia.
Results Neither the score of subtests of the LOTCA nor the overall scores of LOTCA of aphasia patients with different gender and educational background differed (all P>0.05). In different age groups, apart from thinking operation (F=3.373, P=0.016), visuomotor organization (F=3.124, P=0.022), attention (F=3.729, P=0.009) and the total score (F=2.683, P=0.041), there was no difference in terms of the other subtest scores of LOTCA (all P>0.05). In the groups of different length of time with illness, apart from orientation (F=2.982, P=0.039) and attention (F=3.485, P=0.022), the score of other subtests and the total score of LOTCA were not different (all P>0.05). In the groups of different degree of aphasia, apart from attention (F=2.061, P=0.074), both the score of other subtests and the total score of LOTCA differed (all P<0.05).
Conclusion LOTCA might be suitable to assessing the cognitive ability of post-stroke Chinese patients with aphasia.

OBJECTIVETo evaluate the diagnostic value of 3.0 T diffusion-weighted imaging with background suppression (DWIBS) magnetic resonance (MR) for lymph node metastasis in rectal cancer.

METHODSThirty-five patients with rectal cancer who underwent preoperative routine MRI+DWI examination were enrolled in the study and were treated by rectal cancer resection plus lymph node dissection. Metastatic and non-metastatic lymph nodes were confirmed by postoperative pathology. Apparent diffusion coefficient (ADC) values, long-axis and short-axis diameters of lymph nodes were measured. Receiver operating characteristic (ROC) curve was used to assess the diagnostic efficacy of ADC, long-axis and short-axis diameters for differentiating metastatic lymph nodes from non-metastatic lymph nodes.

RESULTSA total of 151 lymph nodes were confirmed with exact location in 35 patients. Sixty-five metastatic lymph nodes and 86 non-metastatic lymph nodes were identified. The ADC values of metastatic lymph nodes and non-metastatic lymph nodes were(0.86±0.14)×10(-3) and (0.94±0.16)×10(-3) mm(2)/s respectively. The long-axis diameter were(9.78±3.13) and (7.90±1.77) mm, respectively. The short-axis diameter were (7.65±2.00) and (6.45±1.19) mm, respectively. There were statistically significant differences between metastatic and non-metastatic lymph nodes in ADC values, long-axis diameter, and short-axis diameter(all P<0.01). The areas under the ROC curve of ADC value, long-axis diameter, and short-axis diameter were 0.648, 0.706, and 0.692, respectively. Optimal cutoff values for these parameters were 1.05×10(-3) mm(2)/s, 7.95 mm, and 5.90 mm, respectively, and the corresponding sensitivities and specificities were 93.8% and 30.2%, 75.4% and 61.6%, 90.8% and 38.4%.

CONCLUSIONSQuantitative measurement of ADC value may reflect the degree of diffusion restriction of metastatic lymph nodes by DWIBS at 3.0 T MR. Accurate diagnosis of metastatic lymph nodes in rectal cancer demands comprehensive evaluation combining ADC value with diameter measurement.

Diffusion Magnetic Resonance Imaging ; methods ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Rectal Neoplasms ; diagnosis ; pathology

Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts- intergenic regions(IGRs), 423 nts-Pseudogene-like sequence (psi), 70 nts-Trailer. The four full-length genomes were in accordance with the properties of Rhabdoviridae Lyssa virus by BLAST and multi-sequence alignment. The nucleotide and amino acid sequences among Chinese strains had the highest similarity, especially among animals of the same species. Of the four full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so the nucleotide mutations happened in these four genomes were most synonymous mutations. Compared with the reference rabies viruses, the lengths of the five protein coding regions had no change, no recombination, only with a few point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the four genomes were similar to the reference vaccine or street strains. And the four strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessed the distinct district characteristics of China. Therefore, these four rabies viruses are likely to be street viruses already existing in the natural world.
Animals ; China ; Deer ; Disease Reservoirs ; virology ; Dogs ; virology ; Ferrets ; virology ; Genome, Viral ; Humans ; Mice ; Molecular Sequence Data ; Phylogeny ; Rabies ; virology ; Rabies virus ; chemistry ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics