Objective To study the regulation effect of estrogen in expression of matrix metalloproteinase-9 (MMP-9) in the central nervous system (CNS) in mice with experimental autoimmune encephalomyelitis ( EAE).Methods The 60 mice were overiectomized and 2 weeks later EAE was induced with MOG35-55 peptide in these mice.They were divided into a treatment group and a control group.The treatment group was treated with estrogen and the control group was given PBS.Clinical symptoms in these two groups were scored and compared.HE staining was used to observe inflammation in the brain and spinal cord.The MMP-9 expression in the CNS was examined by quantitative real-time PCR and immunofluorescence staining.Results The incidence of disease was lower (treatment and control group were 8/30 and 28/30 respectively) and clinical symptoms were milder (treatment and control group were 3.23±0.83 and 1.62 ±1.00 respectively,t=3.811 and P＜0.05) in the treatment group than those in the control group.HE staining showed the decreased infiltration of inflammatory cell in the treatment group (Treatment group:inflammatory score were 0.895 ±0.206,0.752 ±0.302,0.732 ±0.183 in acute,relief and chronic phase respectively;Control group:inflammatory score were 3.472 ±0.635,2.881 ±0.662,1.891 ± 0.482 in acute,relief and chronic phase respectively.t = 8.622,6.543 and 5.027,all P ＜ 0.05).The quantitative real-time PCR and immunofluorescence staining showed that the expression of MMP9 in the CNS was decreased in the treatment group.Conclusion Estrogen may decrease MMP-9 expression in the CNS,reduce inflammation and clinical symptoms in mice with EAE.

Anemia, Aplastic ; physiopathology ; therapy ; Child ; Child, Preschool ; China ; Cord Blood Stem Cell Transplantation ; adverse effects ; methods ; Female ; Graft Survival ; Graft vs Host Disease ; etiology ; immunology ; therapy ; HLA Antigens ; blood ; immunology ; Histocompatibility ; immunology ; Histocompatibility Testing ; Humans ; Male ; Treatment Outcome

BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.

Objective To explore the feasibility of application of multiplex quantitative fluorescent PCR with non-polymorphic loci in prenatal diagnosis of aneuploidies. Methods From Mar 2006 to Nov 2007, a total of 63 samples were collected from the Department of Obstetrics and Gynecology, Affiliated Drum Tower Hospital of Medical College, Nanjing University, including 54 villous samples obtained for karyotyping because of spontaneous abortion, six anmiotic fluid samples of second trimester and three umbilical cord blood samples of third trimester. Blood samples of 60 healthy adults were obtained at the same time as a control group, including 30 males and 30 females. Non-polymorphic QF-PCR was performed on both testing group and control group for the detection of aneuploidies. The Amelogenin gene (AMXY) was selected as an internal control, and dosage quotiety (DQ) of each locus was calculated according to the known formula, ff DQ was between O. 7 and 1.3, the sample was considered as normal If the figure turned out to be >1.3 or <0.7, a potential duplication or deletion of the corresponding gene or chromosome was indicated. If the results implied numerical abnormalities in more than one euchromusome, sex chromosome aneupioidies should be considered. Cell culture and karyotyping were carried out for every sample simultaneously. The results of non-polymorphic QF-PCR were checked with karyotypes. Results ( 1 ) In the control group, all female samples presented only an AMX peak for sex chromosome while all males showed AMX and AMY amplified peaks. The AMY/AMX ratios were between 0.7-1.3, and SD was between 0.05-0.12. (2) Among 19 QF-PCR abnormal cases, 13 cases were proved by karyotyping. Of the six cases which turned out to be conflicting, one case of trisemy 18 shown by karyotyping was not completely detected by QF-PCR, a locus on chromosome 18 implied trisomy, while another turned out to be normal(DQ=1.28). Four cases were detected by non-polymorphic QF-PCR as trisemies but showed normal female karyotype because of maternal contamination during cell culture. A karyotyping]y ' 46, XY' case did not present an AMY peak. Thirty-six out of 44 (82%) normal results implied by non-polymorphic QF-PCR were in accordance with cytogenetic analysis. Of the other eight cases, one case which failed cytogenetic analysis was detected by QF-PCR as normal Four cases showed multiploidy by karyotyping but normal in QF-PCR analysis, including three eases of 69, XXX, one case of 92, XXXX and one case of 45,XX,rob(13;21). The other two cases that showed normal male results turned out to be normal female karyotypes. Conclusions Prenatal aneuploidy detection by non-polymorphic QF-PCR is feasible in a clinical diagnostic setting. With the advantages of high throughput, rapidness and low cost, this method shows a good prospect in clinical application.

Objective To learn the plague's host animals and parasitic flea composition, and to investigate the natural foci of plague in Xiji county in order to provide basic information for plague prevention and control. Methods The Citellus alaschanicus density, nocturnal rodents, the body flea, the burrow track flea, the nest flea were investigated in 8 townships (town) of Xiji county from June 11 2007 to July 25 2007. Specimens of small mammalian, fleas were collected for bacteriological and serological testing. Results The average density of the main host Citellus alaschanicus was 0.85 per hectare. The nocturnal mouse capture rate was 0.80%(24/2987).The survey found 16 species of small mammals that belonging to 3 orders, 9 families and 16 species with Citellus alaschanicus the dominant species. The Citellus alaschanicus had 2.84 fleas per body. Four families and 16 species of fleas were identified in the areas. The Citellus alaschanicus and Citellophilus Tesquorum Mongolicus were the dominant species. Plague bacteriology and serology tests were negative. Conclusions The study shows that the area is suitable for the formation of natural foci of Citellus alaschanicus plague. Surveillance is an important measure for prevention and control of the plague.

Objective To study the biodistribution of anti-HER-2/neu monoclonal antibody Herceptin labeled by 131I(131I-Herceptin) in healthy KM mice and nude mice bearing human ovarian cancer xenografts and radioimmunoimaging (RII) of the nude xenografts-bearing mice.Methods 131I-Herceptin was prepared using Iodogen method.The labeling efficiency, radiochemical purity, stability and immunocompetence were measured.The percentage activity of injection dose per gram of tissue (%ID/g) and the radioactivity ratio of tumor to non-tumor tissue (T/NT) were calculated for each time point.The optimal time for imaging was investigated by comparing the 131I-Herceptin SPECT for the nude mouse models bearing ovarian cancer xenografts at different time points.Results The labeling efficiency and radiochemical purity of 131I-Herceptin were 89.8% and 98.4%, respectively.The labeling was stable and had good immunocompetence.131 I-Herceptin was cleared rapidly mainly through liver, spleen and kidneys, consistent with first order two-compartment model.The uptake of 131I-Herceptin in the tumors bearing human SKOV-3 xenografts was much higher than that in nontumor tissue.The% ID/g was 18.08 in the tumor at 24 h post injection.The T/NT ratio increased with time and was 27.27 at 72 h post injection.The tumors in nude mice bearing SKOV-3 xenografts could be visualized on 131I-Herceptin SPECT imaging 2 h post injection; definitely identiffed 48 h post injection and the radioactivity ratio of tumor to contralateral tissue was 11.44 at 120 h post injection.However, the tumor in nude mice bearing HO-8910 xenografts did not show abnormal uptake of 131 I-Herceptin at each time point.Conclusions 131 I-Herceptin is a good radiopharmaceutical targeting SK-OV-3 xeuografts and it may be useful in imaging carcinoma of ovary and target therapy of its metastases with high HER-2/neu expression.

Objective To analyze clinical and pathological features of dermatomyositis with panniculitis as a skin manifestation.Methods Clinical data were collected from 9 cases of dermatomyositis with panniculitis as a skin manifestation in Department of Dermatology of Zhongshan Hospital affiliated to Fudan University from October 2012 to July 2016,and their clinical and pathological features were analyzed.Results Of the 9 cases,6 were female and 3 were male,and the age ranged from 28 to 73 years.Panniculitis lesions of the 9 patients all manifested as painful indurated plaques or nodules on the buttock,thigh,waist,back,abdomen,upper extremities and cheeks.These lesions occurred before,after or simultaneously with the onset of characteristic skin and muscle lesions of dermatomyositis,especially preceded the onset of characteristic lesions of dermatomyositis by 30 years in 1 case.Histopathological examination of lesions showed liquefaction degeneration of basal cells,inflammatory infiltration of lymphocytes and plasma cells around blood vessels,in the fat lobules as well as between the lobules and septa in the dermis.The necrosis and calcification of lipocytes,lipomembranous changes,fibrinoid necrosis of damaged vessel walls and microvascular occlusion were observed in some cases.Because panniculitis preceded the onset of characteristic lesions of dermatomyositis,2 patients were misdiagnosed with lupus panniculitis and morphea profunda for several times.Most patients had good response to systemic glucocorticoids combined with immunosuppressive agents,while the patients with lipomembranous fat necrosis had poor response to the combination therapy.Conclusions Panniculitis lesions of dermatomyositis are histologically characteristic,and may do not coincide with the onset of characteristic lesions of dermatomyositis.If panniculitis lesions precede characteristic lesions of dermatomyositis,patients will be easily misdiagnosed.Thus,persistent follow-up visit will be of great importance for the diagnosis.

Objective To investigate the value of platelet-to-lymphocyte ratio (PLR) in the prognosis prediction of patients with diabetic ketoacidosis (DKA).Methods Total of 105 patients with DKA who were treated in resuscitation room of Peking Union Medical College Hospital from January 1,2006 to December 31,2015 were reviewed.Among them,there were 8 cases died,and the other 97 cases survived.Another 105 patients with diabetes mellitus who were treated in the ward of Endocrinol ogy Department in the same period were selected as non DKA control group.The clinical characteristics of the patients in each group were compared and Logistic regression analysis was performed on the prognosis of DKA.Results Mechanical ventilation,simultaneous other organ dysfunction,PLR,Glasgow coma score related to prognosis of DKA (P ＜ 0.05).The OR value of platelet-to-lymphocyte ratio was 3.242.The optimal cutoff value of PLR for predicting the prognosis of patients was 256.50.Its sensitivity and specificity were 87.5％ and 88.7％,respectively.Conclusions PLR can be used as a sensitive indicator to predict the prognosis of DKA patients.

Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P＜0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P＜0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P＞0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P＜0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P＜0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.

Objective To investigate the antimicrobial resistance of clinical isolates in Tongling People′s Hospital during 2013. Methods A total of 2 281 nonduplicate clinical isolates were collected.Kirby-Bauer disc diffusion method was employed to study the antimicrobial susceptibility.The data were analyzed with WHONET 5.6 software according to CLSI 2012 breakpoints. Results The top 5 most frequently isolated microorganisms were E.coli (479,21.0%),K.pneumoniae (360,15.8%),A. baumannii (271,11.9%),P .aeruginosa (240,10.5%),S.aureus (171,7.5%).Gram negative and gram positive microorganisms accounted for 76.5% and 23.5%,respectively.The prevalence of methicillin-resistant strains in S.aureus (MRSA)and coagulase negative Staphylococcus (MRCNS)was 38.6% and 73.1%,respectively.The resistance rates of MR strains to beta-lactams and other antimicrobial agents were much higher than those of MS strains.No staphylococcal strain was found resistant to vancomycin or teicoplanin.E.faecalis showed relatively lower resistance to penicillin,ampicillin and nitrofurantoin.E.faecium strains were more resistant than E.faecalis to most of the antibiotics tested.Approximately 50.5% of E.coli and 44.5% of Klebsiella isolates produced extended-spectrum beta-lactamases (ESBLs).The ESBLs-respectively.And 29.8% and 23.4% of the P .aeruginosa strains were resistant to imipenem and meropenem.Nearly all (94.0%)P .aeruginosa isolates were susceptible to amikacin.Conclusions There appears a trend of increasing resistance in the clinical bacterial isolates in this hospital,especially the carbapenem-resistant Enterobacteriaceae,which is of great concern.It is mandatory to take effective antibiotic policy and infection control measures.