Background Expulsive choroidal haemorrhage is a severe complication during the intraocular operation.To create a feasible animal model of expulsive choroidal haemorrhage is very important for the study of its nature disease course and therefore prevention and treatment.Objecfive Present study was to modify the method of establishing acute choroidal haemorrhage animal model and explore the risk factor and pathogenesis mechanism.Methods The suprachoroidal explusive haemorrhage animal models were created by modifying the Zauberman method to ligate 2-4 vortex veins and rapidly intravenous injection of 250 ml normal solution to elevate the intraocular pressure(IOP),and then low the IOP by opening the anterior chamber via limbal incision.The bleeding of choroid and detachment of retina were examined by B-type ultrasound immediately,1,2 and 4 weeks after operation to assess the disease alteration with lapse of time.The samples of eyeballs were prepared in mentioned above time points for the histopathologieal evaluation of retina tissue.Results The animal models of suprachoroidal explusive haemorrhage were established bv this modified method with the successful rate 100％,and the manifestation resembled a clinical disease course.The shape,range and location of choroidal bleeding and retinal detachment were determined,and the diagnosis and change of suprachoroidal explusive haemorrhage with time were clarified using B-uhrasonography.The histopathological change revealed the breaking of Brueh's membrane and retinal pigmentary layer.It was verified that the dominant pathological change of retina was retinal destroy in 1 week after operation,and in 4 weeks after operation,the main pathological mechanism was proliferation of fibroblasts and collagen fiber. Conclusion The consequence procedure of modified Zauberman method,quickly intravenous injection of normal solution and puncture of the anterior chamber can create an ideal animal model of suprachoroidal explusive haemorrhage.The rapid IOP descent is a main risk factor of supraehoroidal explusive haemorrhage.The results of supraehoroidal explusive haemorrhage is severe destroy of retina and ehoroid.This study implies that supraehoroidal explusive haemorrhage is likely to be prevented and treated in the early stage.

Objective To figure out changes of serum excitatory amino acids (EAAs) levels in patients with amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD).Methods The levels of serum EAAs was assessed in 34 cognitively normal control subjects,30 patients with aMCI,and 32 patients with AD using high performance liquid chromatography (HPLC).Results ①Higher serum concentrations of glutamate((39.6?22.1) ?mol/L),alanine((282.5?71.3) ?mol/L) were found in the aMCI patients (P=0.044,P=0.007),and higher serum concentrations of glutamate ((42.2?21.8) ?mol/L),glycine ((464.2?142.6) ?moL/L) were found in the AD patients than in the control subjects (P=0.010,P=0.010).②No statistically significant difference of EAAs level between the aMCI and AD groups was found.③A close and positive correlation between the serum concentrations of glutamate, aspartate and the mini-mental status examination scores were found in AD patients:the 2 amino acid levels were higher in patients with mild dementia((42.1?21.3),(55.0?29.0) ?mol/L) than those with moderate or severe dementia ((25.4?9.2) ?mol/L,P=0.023;(34.6?11.1) ?mol/L,P=0.036). Conclusion EAAs,correlating with the severity of the condition,play a significant role in AD,while aMCI patients also have disturbance of metabolism of EAAs,indicating that it has similar pathogenesis to AD.

Objective To study the clinical features of diagnosis and treatment in children with low-risk myelodysplastic syndrome(MDS) and improve the diagnosis and treatment.Methods The examinations of 17 children with low-risk MDS were analyzed.The blood concentrations of cyclosporine,one of the therapeutic drugs,were monitored and the responses to the treatments were evaluated.Results Exa-mination of full blood count showed that the reductions of 3 cell types,2 cell types and 1 cell type were 11 (64.7%)cases,5(29.4%)cases and 1(5.9%)case,respectively.Reticulocyte count showed an increase in 82.4% of the patients and normal in 3 cases.Fourteen in 17 cases were hyperplastic marrow and 3 cases were hypoplastic marrow.Among all cases,one lineage,2 lineages and 3 lineages dyspoiesis were seen in 8(47.1%),7(41.2%) and 1 (5.9%)cases,respectively.One case showed no dyspoiesis.Cytogenetics examination showed normal in 10(58.8%) cases and abnormal in 7(41.2%) cases.Fifteen (88.2%) cases had normal proportions of CD59 negative cells,while 2 cases had higher proportions.The blood concentrations of cyclosporine that were tested in 9 cases at the end of the third week were in a range of 95.3-316.5 ?g/L.The therapeutic effect of 10 cases were evaluated at the end of the third month after being treated.Eight cases achieved haematological improvement and 2 cases didn′t.The rate of improvement was 80%.Conclusions The patients of low-risk MDS are mostly school-aged children and pancytopenia is the most common sign.The combination of predisone,cyclosporine and stanozolol agents shows good effect to treat low-risk MDS.The absorption of cyclosporine is different individually,so it is significant to adjust the dosage of cyclosporine according to the concentration regularly in clinical practice.

Objective To explore the clinical characteristics of nosocomial infection in children with acute leukemia and the strategy of prevention and treatment.Methods One hundred and thirty-three cases of nosocomial infection in children with acute leukemia were analyzed by retrospective study.The relationship between nosocomial infection and stage of leukemia,hospitalization duration,and the rate of infection were investigated.Results Nosocomial infection rate was 53.4%(71/133 cases),significant difference of infection rate between acute lymphoblastic leukemia and nonlymphoblastic leukemia group was found(P0.05).The main pathogens of septicaemia were gram negative bacilli,and they were generally sensitive to Amicacin and Pi-peracillin/tazobactam.Conclusions Children with acute leukemia have high nosocomial infection rate.The occurrence of nosocomial infection was related to the type and stage of leukemia and hospitalization duration but not to the prognosis.The main pathogens of septicaemia were gram negative bacilli.

Human leukocyte antigen G (HLA-G), a kind of non-classical major histocompatibility complex class I antigens, can inhibit inflammatory reaction, assist tumor cells to escape from immune surveillance and promote the immunologic tolerance of the graft. HLA-G, expressed and secreted by human amniotic mesenchymal cells (HAMC), suppresses the functions of NK cells, T cells and B cells and modulates the activity of dendritic cells (DC). These findings provide a theoretical basis for illustrating the mechanism of immunosuppression on HAMC. In this article, the recent advances on not only the gene and the molecular structure of HLA-G, but also the possible mechanisms of HLA-G in immunoregulatory function of HAMC, as well as the relation of HLA-G with HAMC, NK, DC, T and B cells are reviewed.
Amnion ; cytology ; HLA-G Antigens ; immunology ; Humans ; Immune Tolerance ; Mesenchymal Stromal Cells ; cytology ; immunology

The aim of this study was to isolate, cultivate and phenotypically characterize two types of human amnio-tic membrane (HAM)-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells (hAEC) were derived from the embryonic ectoderm, while human amnion mesenchymal cells (hAMC) were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immunofluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers (CD29, CD73, CD44, CD59, CD90, CD105, CD166), but did not express the hematopoietic markers (CD31, CD34, CD45, HLA-DR) and showed the weak expression of costimulatory molecules (CD40, CD40L, CD80, CD86). Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytes potential, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.
Amnion ; cytology ; Cell Differentiation ; physiology ; Cell Lineage ; Epithelial Cells ; cytology ; Humans ; Immunophenotyping ; Stromal Cells ; cytology

Objective： This study was designed to investigate the relationship between expression level of multidrug resistance-associated protein (MRP) or P-glycoprotein (P-gp) genes and chemotherapy efficacy or clinical drug resistance in the patients with malignant lymphomas. Methods： Using the methods of semi quantitative reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry (FCM), The authors examined the expression level of MRP and P-gp of 46 lymphoma patients. Results： The expression level and positive rate of P-gp in the recurrent patients was higher than that in untreated patients (P<0.001). There was no difference in MRP gene expression level and positive rate between recurrent and untreated patients (P>0.05). Chemotherapeutic effective rate in P-gp positive patients (26.67％ ) was lower than that in P-gp negative patients (83.87％ )(P<0.001). While there was no difference between MRP gene positive and negative patients, their chemotherapy effective rate had no difference. Relevant analysis showed that there was no correlation between MRP and P-gp (r-0.0808, P>0.05). Conclusion： P-gp expression correlates to multidrug resistance, and is the major mechanism of clinical drug resistance of lymphomas, whereas, MRP gene does not appears to play any role in that course.

OBJECTIVEThe study was aimed to investigate the feasibility and clinical significance of quantitative detection of minimal residual disease (MRD) in childhood acute lymphoblastic leukemia (ALL) by real-time quantitative polymerase chain reaction (RQ-PCR).

METHODSClonal IgH gene rearrangements of samples at diagnosis were identified by standard PCR assay with consensus primers. Monoclonal IgH gene rearrangements were analyzed using DNAPLOT software. Upstream primers were designed with the Primer Express software and allele specific oligonucleotide developed complementary to the V-D or D-J junction. Samples at diagnosis were serially diluted to generate the patient specific standard curves. RQ-PCR method was used to quantify the MRD of the follow up samples collected at five time points during chemotherapy. To check the quantity and quality of DNA, the investigators used RQ-PCR analysis for the albumin gene.

RESULTSTotally 16 monoclonal IgH gene rearrangements were identified from 34 patients with B-ALL. The analysis of the 16 monoclonal rearrangements showed that the most frequently used V segment was from V3 family and J segment from J4 and J6. The RQ-PCR sensitivity of 10(-4) to 10(-5) was mostly reached. Non-specific amplification was seen in 6 patients. The number of inserted and deleted nucleotides did not appear to be related to the sensitivity (P > 0.05). The correlation coefficients of all 16 standard curves were excellent (> or = 0.99). The mean slope of the standard curves was -3.4 +/- 0.37 and the mean intercept was 24.3 +/- 2.95. MRD analysis of follow up samples from the 16 patients showed an association between high degree of MRD and relapse. There was no apparent relationship between MRD degree at the end of induction chemotherapy and other high risk factors of ALL (P > 0.05).

CONCLUSIONThe study showed that the above approach with RQ-PCR was applicable to clinical detection of MRD in childhood ALL. Quantitative and dynamic study of MRD was of prognostic importance.

Child ; Gene Rearrangement, B-Lymphocyte ; Humans ; Neoplasm, Residual ; diagnosis ; genetics ; Polymerase Chain Reaction ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; drug therapy ; genetics ; Prognosis

Animals ; Cattle ; Chickens ; Escherichia coli Infections ; epidemiology ; microbiology ; Escherichia coli O157 ; isolation & purification ; pathogenicity ; Sheep ; Swine

OBJECTIVETo study the effect of ursolic acid (UA), apentacyclic triterpene acid, on MCF-7 cell apoptosis, and probable mechanism involved by detecting the expressions of caspase-3 and poly ADP-ribose polymerase(PARP) at protein level.

METHODMCF-7 cells were cultured with different concentrations of UA. Growth inhibition of UA on MCF-7 cells was evaluated by MTT assay. Cell cycle and sub-G1 peak were performed by FCM. Morphologic changes of UA-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Ho 33258 staining by a fluorescence microscope (EX: U. V.). The protein expression of caspase-3 and PARP was analyzed by immunofluorescence cell staining (SABC-Cy3).

RESULT24 hours after UA treatment, inhibition of MCF-7 cell growth was concentration-dependent. The IC50 value for UA was (22.6 +/- 3.0) micromo x L(-1). Cell cycle anaysis by FCM showed that 50 micromol x L(-1) of UA arrested MCF-7 cell cycle at G0 - G1 phase. Morphological changes of MCF-7 Cells exhibited many of the hallmark features of apoptosis, including chromatin clumps and aggregation and DNA fragmentation. UA increased caspase-3 protein expression.

CONCLUSIONThe results suggest that UA evokes MCF-7 cell apoptosis is correlation with the up-regulation of caspase-3. Our study indicated that UA might be a potential Chinese medical component for breast neoplasm.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; enzymology ; pathology ; Caspase 3 ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Female ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Triterpenes ; pharmacology