ObjectiveTo develop a stable model of focal cerebral infarction in rat to study the curative effect of neural stem cells transplantation.MethodsThirty-seven rats were selected which were divided into two groups in random, experimental group and control group. The focal infarction model was developed by the ligation of the left middle cerebral artery followed by the ligation of the ipsilateral common carotid artery and the temporary clip occlusion of the contralateral common carotid artery for 1.5 h. The operation adopted minimally invasive craniotomy though temporal bone. The model was evaluated by examining the neurologic deficits, ink perfusion, TTC staining and Magnetic Resonance imaging.ResultsAll the rats were in good condition after the operation, the mortality rate was 6.25% after 4 weeks. Ink perfusion and TTC staining confirmed that the ischemia was confined to the cortex. The areas of infarction measured 83.52 mm3 by Magnetic Resonance imaging after 4 weeks.ConclusionA stable focal cerebral infarction model can be achieved by minimally invasive craniotomy. It is superior for its homogeneity of infarction volume and site, and its low mortality. It can be used for the study of transplantation of neural stem cells.

Neurotrophins promote and modulate the repair and regeneration of central nervous system (CNS) on the levels of synapses, neurites and neural cells, and even of the auxiliary structures of CNS. As to immune molecules, recent studies denied the classical doctrine that CNS is an immune privilege organ. In the last decade, it is found that immune responses can be beneficial for CNS repair. What are more, some macromolecules that were previously thought to be the members of the family of immune system play essential roles in the development and regeneration of the CNS. Therefore, the authors postulate that there are crosstalks between the neurotrophins and the immune molecules in the CNS.

BACKGROUND: Nanoparticles (NPs) prepared from biodegradable polymers, such as poly (D,L-lactic acid-co-glycolic acid) (PLGA), have been studied as vehicles for the delivery of antigens to phagocytes. This paper describes the preparation of antigen-loaded PLGA-NPs for efficient cross-priming. METHODS: NPs containing a similar amount of ovalbumin (OVA) but different sizes were produced using a micromixer-based W/O/W solvent evaporation procedure, and the efficiency of the NPs to induce the cross-presentation of OVA peptides were examined in dendritic cells (DCs). Cellular uptake and biodistribution studies were performed using fluorescein isothiocyanate (FITC)-loaded NPs in mice. RESULTS: The NPs in the range of 1.1~1.4microm in size were the most and almost equally efficient in inducing the cross-presentation of OVA peptides via H-2Kb molecules. Cellular uptake and biodistribution studies showed that opsonization of the NPs with mouse IgG greatly increased the percentage of FITC-positive cells in the spleen and lymph nodes. The major cell type of FITC-positive cells in the spleen was macrophages, whereas that of lymph nodes was DCs. CONCLUSION: These results show that IgG-opsonized PLGA-NPs with a mean size of 1.1microm would be the choice of biodegradable carriers for the targeted-delivery of protein antigens for cross-priming in vivo.
Animals ; Cross-Priming ; Dendritic Cells ; Fluorescein ; Immunoglobulin G ; Isothiocyanates ; Lactic Acid ; Lymph Nodes ; Macrophages ; Mice ; Nanoparticles ; Ovalbumin ; Ovum ; Peptides ; Phagocytes ; Polyglycolic Acid ; Polymers ; Spleen

Previously we showed that biodegradable nanoparticles containing poly-IC or CpG oligodeoxynucleotide (ODN) together with ovalbumin (OVA) were efficient at inducing MHC-restricted presentation of OVA peptides in dendritic cells. The CTL-inducing activities of the nanoparticles were examined in the present study. Nanoparticles containing poly-IC or CpG ODN together with OVA were prepared using biodegradable polymer poly(D,L-lactic acid-co-glycolic acid), and then were opsonized with mouse IgG. The nanoparticles were injected into the tail vein of mice, and 7 days later the OVA-specific CTL activities were measured using an in vivo CTL assay. Immunization of mice with the nanoparticles containing poly-IC or CpG ODN together with OVA elicited potent OVA-specific CTL activity compared to those containing OVA only. In accordance with these results, nanoparticles containing poly-IC or CpG ODN together with OVA exerted potent antitumor activity in mice that were subcutaneously implanted with EG7.OVA tumor cells. These results show that encapsulation of poly-IC or CpG ODN together with antigen in biodegradable nanoparticles is an effective approach for the induction of potent antigen-specific CTL responses in vivo.
Animals ; Dendritic Cells ; Immunization ; Immunoglobulin G ; Lactic Acid ; Mice ; Nanoparticles ; Ovalbumin ; Ovum ; Peptides ; Polyglycolic Acid ; Polymers ; Veins

BACKGROUND: Enterovirus is a common cause of aseptic meningitis, respiratory disease and nonspecific febrile illness. The conventional methods for laboratory diagnosis of enterovirus infections have been virus culture and serotyping by an immunofluorecent test. We studied a new and more rapid approach for enterovirus detection in cerebrospinal fluid (CSF) by real-time nested PCR. METHODS: This study was performed on 50 CSF specimens from patients suspected of aseptic meningitis. Enterovirus was detected in CSF by PCRs for 3 different targets and real-time nested PCR. Enterovirus culture was also performed in 44 CSF specimens. RESULTS: The positive rate of PCRs for each of the 3 different targets was 26.0%, 40.0%, or 46.0%, and that of real-time nested PCR was 86.0%. Only 6.8% were positive in culture. Thus, the positive rate of real-time nested PCR was much higher than other methods. CONCLUSIONS: Our study revealed that the real-time nested PCR should be useful for diagnosis of enterovirus infections because of a high sensitivity and rapid detection.
Cerebrospinal Fluid* ; Clinical Laboratory Techniques ; Diagnosis ; Enterovirus Infections ; Enterovirus* ; Humans ; Meningitis, Aseptic ; Polymerase Chain Reaction* ; Real-Time Polymerase Chain Reaction ; Reverse Transcription* ; Serotyping

Autoimmune hepatitis in children is a rare and severe inflammatory disease of unknown etiology, and progress to cirrohosis and liver failure, generally is responsive to immunosuppressive therapy. It is more prevalent in women than men, and characterized by the presence of circulating autoantibodies, a high serum globulin. Extrahepatic manifestations such as thyroiditis, ulcerative colitis, glomerulonephritis and autoimmune hemolytic anemia, are associated. We report, to our knowledge, the first case of autoimmune hepatitis in conjunction with choledochal cyst and pancreatitis in 11-year-old female patient. At the time of diagnosis, she suffered from acute upper abdominal pain, jaundice, and pallor. Laboratory findings showed Cooms positive hemolytic anemia, hypergammaglobulinemia, hyperbilirubinemia, and high serum transaminases. Antinuclear antibody was of homogeneous type. In liver biopsy, cellular infiltrates largely lymphocytes were noted. Treatment with corticosteroids induced clinical, biochemical remission, but subsequent withdrawal leaded to relapse. Incidentally choledochal cyst were found and then acute pancreatitis developed. After management for acute pancreatitis, surgical resection of cyst with hepatojejunostomy was performed.
Abdominal Pain ; Adrenal Cortex Hormones ; Anemia, Hemolytic ; Anemia, Hemolytic, Autoimmune ; Antibodies, Antinuclear ; Autoantibodies ; Biopsy ; Child ; Choledochal Cyst* ; Colitis, Ulcerative ; Diagnosis ; Female ; Glomerulonephritis ; Hepatitis, Autoimmune* ; Humans ; Hyperbilirubinemia ; Hypergammaglobulinemia ; Jaundice ; Liver ; Liver Failure ; Lymphocytes ; Male ; Pallor ; Pancreatitis* ; Recurrence ; Thyroid Gland ; Thyroiditis ; Transaminases

OBJECTIVETo evaluate the quantitative and functional changes of T helper (Th) cell subsets in the bone marrow of severe aplastic anemia (SAA) patients and the relationship between these changes and the patients hematopoietic function.

METHODSBy FACS, the quantity and ratio of Th1 and Th2 cells, the percentage of CD3(+)CD8(+) cells in the bone marrow were detected in 24 patients with SAA at active phase, 15 patients with SAA at recovery phase, and 16 normal controls. By radioimmunoassay, the serum levels of TNF-alpha, or IL-4 in 20 SAA patients at active phase, 12 at recovery phase and 16 normal controls were measured. The relationships between CD3(+)CD8(+) cells, TNF-alpha and Ret, ANC; and between Th1 cells and CD3(+)CD8(+) cells, TNF-alpha or Ret, ANC; between IL-4, balance of Th1/Th2 and Ret, ANC were evaluated.

RESULTSThe percentages of Th1 and Th2 cells, and ratio of Th1/Th2 in bone marrow of SAA patients at active phase were (4.87 +/- 2.64)%, (0.41 +/- 0.26)% and 21.22 +/- 5.07, respectively, being higher than those of normal controls [(0.42 +/- 0.30)% (P < 0.01), (0.24 +/- 0.17)% (P < 0.05) and (1.57 +/- 0.93) (P < 0.01), respectively] and all of them reduced to normal levels of SAA at recovery phase (P > 0.05). The percentage of CD3(+)CD8(+) cells significantly decreased from (32.32 +/- 8.69)% at active phase to (13.76 +/- 2.96)% at recovery phase (P < 0.01). The serum levels of TNF-alpha and IL-4 at active phase was (4.29 +/- 3.15) microg/L and (1.24 +/- 0.73) microg/L, respectively, being higher than those of normal controls (1.21 +/- 1.16) microg/L, (1.18 +/- 0.97) microg/L, but only the difference of TNF-alpha was statistically significant (P < 0.01). In recovery SAA patients, the serum levels of TNF-alpha significantly decreased to (1.46 +/- 1.41) microg/L (P < 0.01), and the levels of IL-4 increased markedly to (3.05 +/- 1.94) microg/L. The CD3(+)CD8(+) cells and TNF-alpha of patients negatively correlated with Ret (P < 0.05; P < 0.05) and ANC (P < 0.05; P < 0.05), Th1 cells correlated with CD3(+)CD8(+) cells and TNF-alpha positively (P < 0.01; P < 0.05), the Ret and ANC negatively (P < 0.01; P < 0.01), IL-4 and the balance of Th1/Th2 positively correlated with Ret and ANC (P < 0.05, P < 0.01; P < 0.01, P < 0.01).

CONCLUSIONThe bone marrow failure in SAA might be caused not only by the increase of Th1 cells, Th1 type effector cells and cytokines, but also by insufficient compensation of Th2 cells and Th2 type cytokines, which shifted the balance of Th1/Th2 favorable to Th1.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; pathology ; physiopathology ; Bone Marrow ; metabolism ; pathology ; CD3 Complex ; blood ; CD8 Antigens ; blood ; Child ; Female ; Hematopoietic System ; metabolism ; pathology ; physiopathology ; Humans ; Interleukin-4 ; blood ; Male ; Middle Aged ; Radioimmunoassay ; T-Lymphocytes, Helper-Inducer ; metabolism ; pathology ; Th1 Cells ; metabolism ; pathology ; Th2 Cells ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

Lymphangioma is a tumor which is composed of lymph vessels and spaces containing lymph. It consists of endothelial cells and supporting tissue elements of the lymphatic system which are involved in the neoplastic process. Grossly, the cysts are thin walled and multiloculated. Intra-abdominal omental origin lymphangiomas are quite rare and only 21 cases were reported in English literature by 1978. The most common location is the mesentery, followed by the omentum, mesocolon, and retroperitoneum. We report a case of huge cystic lymphangioma originating from the greater omentum in a 14-year-old male patient with brief review of literatures.
Adolescent ; Endothelial Cells ; Humans ; Lymphangioma ; Lymphangioma, Cystic* ; Lymphatic System ; Male ; Mesentery ; Mesocolon ; Omentum*

OBJECTIVETo study the transplantation efficacy of neural stem cells (NSCs) and Schwann cells (SC) in a rat model of spinal cord contusion injury.

METHODSMultipotent neural stem cells (NSCs) and Schwann cells were harvested from the spinal cords of embryonic rats at 16 days post coitus and sciatic nerves of newborn rats, respectively. The differential characteristics of NSCs in vitro induced by either serum-based culture or co-culture with SC were analyzed by immunofluorescence. NSCs and SCs were co-transplanted into adult rats having undergone spinal cord contusion at T9 level. The animals were weekly monitored using the Basso-Beattie-Bresnahan locomotor rating system to evaluate functional recovery from contusion-induced spinal cord injury. Migration and differentiation of transplanted NSCs were studied in tissue sections using immunohistochemical staining.

RESULTSEmbryonic spinal cord-derived NSCs differentiated into a large number of oligodendrocytes in serum-based culture upon the withdrawal of mitogens. In cocultures with SCs, NSCs differentiated into neuron more readily. Rats with spinal cord contusion injury which had undergone transplantation of NSCs and SCs into the intraspinal cavity demonstrated a moderate improvement in motor functions.

CONCLUSIONSSC may contribute to neuronal differentiation of NSCs in vitro and in vivo. Transplantation of NSCs and SCs into the affected area may be a feasible approach to promoting motor recovery in patients after spinal cord injury.

Animals ; Cells, Cultured ; Disease Models, Animal ; Female ; Kaplan-Meier Estimate ; Motor Activity ; Neurons ; cytology ; transplantation ; Postoperative Period ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Schwann Cells ; transplantation ; Spinal Cord ; pathology ; Spinal Cord Injuries ; chemically induced ; therapy ; Stem Cell Transplantation ; Stem Cells ; cytology

Objective To study the correlation of neural cell adhesion molecule 1 (NCAM1) with pathogenesis of cognitive dysfunction in epileptic rats and explore its effect on cognitive dysfunction in epileptic rats.Methods A total of 120 Wistar rats were randomly assigned to control group (n=20) and experimental group (n=100); and rats in the experimental group were subdivided into 5 groups (n=20),the epilepsy models were induced by pilocarpine,and rats in these 5 groups received daily treatments for 30 days with either saline,carbamazine,oxcarbazine,aniracetam or donepezil (nootopic).Spatial learning and memory abilities were assessed with Water Maze test.Hippocampus tissue was assessed for NCAM1 mRNAs by RT-PCR and proteins by immunochemistry.Results The mean escape latency of rats in the Morris water maze test:carbamazepine treatment group ＞oxcarbazepine treatment group ＞aniracetam treatment group ＞epilepsy group ＞donepezi treatment group; significant differences were noted between each two groups (F=91.920,P=0.000).Immunohistochemical and RT-PCR results:donepezi treatment rgroup ＞aniracetam treatment group ＞epileptic group ＞oxcarbazepine treatment group ＞carbamazepine treatment group＞ control group; there were differences between each 2 groups (Immunohistochemical result:F=324.510,P=0.000; RT-PCR result:F=81.160,P=0.000).Conclusion NCAM1 expression in the hippocampus rises within 30 days of epileptic attack,who participates in the pathogenesis of cognitive dysfunction; carbamazepine can aggravate the cognitive dysfunction and donepezil can obviously improve the cognitive function of epilepsy.