OBJECTIVETo study the factors influencing short-term prognosis of tuberculous meningitis (TBM) in children.

METHODSThe clinical data of 137 hospitalized children with TBM between January 2007 and February 2011 were retrospectively reviewed. A total of 30 potential factors influencing short-term prognosis of TBM were evaluated by univariate analysis and multivariate logistic regression analysis.

RESULTSClinical staging showed that of the 137 children 21 cases (15.3%) were in the early stage, 67 cases (48.9%) in the medium stage and 49 cases (35.8%) in the late stage of TBM. The univariate analysis revealed 8 factors associated with a poor short-term prognosis: clinical stage of TBM (late), coma, positive Babinski signs, cranial nerve involvements, paralysis, seizures, obvious abnormalities in brain computed tomography (CT) or magnetic resonance imaging (MRI) and elevated protein concentrations in cerebrospinal fluid (CSF). Factors associated with a favourable short-term prognosis for TBM included glucocorticoid steroids therapy, positive reaction of PPD skin test and an increased length of stay in hospital. Multivariate logistic analysis revealed two independent risk factors for a poor short-term prognosis: clinical stage of TBM (late) (OR: 11.168, 95%CI: 3.521-35.426) and positive signs of meningeal irritation (OR: 4.275, 95%CI: 1.043-17.521). An increased length of stay in hospital was shown as a favorable factor (OR: 0.893, 95%CI: 0.825-0.968).

CONCLUSIONSLate-stage TBM and positive signs of meningeal irritation suggest a poor prognosis, while an appropriately longer length of stay in hospital may contribute to a favorable short-term prognosis for children with TBM.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Prognosis ; Retrospective Studies ; Tuberculosis, Meningeal ; complications ; diagnosis

Humans ; Subacute Sclerosing Panencephalitis

ObjectiveTo understand the distribution of interleukin(IL)-28B gene polymorphisms in human immunodeficiency virus (HIV)/hepatitis C virus (HCV) coinfected Han patients in Hubei Province.MethodsOne hundred Han patients with anti-HIV and anti-HCV double positive in Hubei Province were enrolled.HCV RNA level was detected by fluorescence quantitative polymerase chain reaction (PCR) and genotyping of IL-28B gene was pcrformed by sequencing.The data were analyzed by chi square test.ResultsThe proportion of IL-28B C/C genotype was 95.0 ％ in target population,arnong which 21.1％ (20/95) were HCV RNA negative.While there were no HCV RNA negative cases in C/T and T/T genotypes (0/5;x2 =1.043,P=0.588).ConclusionAmong HIV/HCV coinfccted Han patients in Hubei Province,the proportion of IL-28B C/C genotype is high.

Objective To explore the application value of ultrasound-guided fine needle aspiration biopsy(FNAB) in diagnosing thyroid nodules of TI-RADS 4-6 categories.Methods A retrospective analysis was made on the pathologic data of 494 patients (totally 501 nodules) who underwent TI-RADS ultrasonographic stratification and ultrasound-guided FNAB.168 thyroid nodules of TI-RADS 4-6 categories were selected from 163 patients and classified into three groups:group T,TI-RADS 4A categories;group Ⅱ,TI-RADS 4B category; group Ⅲ,TI-RADS 5-6 category(the pathologically confirmed cases in category 6 had been removed).The ultimate pathologic findings were taken as golden standards to calculate the accuracy,sensitivity,specificity,positive predictive value of ultrasound-guided FNAB in the three groups.ROC curve was drawn to evaluate the diagnostic value of FNAB.Results The accuracy,sensitivity,specificity,positive predictive value of the ultrasound-guided FNAB in the three groups were listed as follows:group Ⅰ 74.4％,73.3％,75.0％,64.7％;group Ⅱ 83.9％,87.5％,71.4％,91.3％;group Ⅲ 89.3％,91.7 ％,75.0％,95.7 ％ respectively.The positive predictive values of the ultrasound-guided FNAB for group Ⅰ and group Ⅱ was higher than that of conventional TI-RADS classification criteria,so the difference was statistically significant; while the positive predictive values for group Ⅲ was close to the conventional ultrasound TI-RADS classification criteria,the difference was not statistically significant.Conclusions The combination application of TI-RADS classification criteria and FNAB can improve the accuracy of ultrasound diagnosis in group Ⅰ and group Ⅱ,and reduce the puncture rate of group Ⅲ.

OBJECTIVETo construct recombinant plasmids containing the truncated gene of the major surface antigen sta56 of Orientia tsutsugamushi (Ot.) Karp strain for expression antigen in E. coli so as to compare the expression efficiency in different systems.

METHODSFrom the recombinant plasmid TOPO-sta56 containing sta56 of Orientia tsutsugamushi Karp strain, several truncated genes of sta56 with different length were amplified and subcloned into the expression vectors pPROEX HTb and pET30a. These genes were expressed in E. coli DH5alpha and BL21(DE3) respectively when induced by IPTG. The expressed recombinant proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot.

RESULTSSix recombinant plasmids containing truncated sta56 genes of different length were constructed as follow: pHTbOt957, pHTbOt498, pHTbOt342 and pETOt957, pETOt498, pETOt342. The recombinant sta56 proteins were highly expressed as 6 x His fusion proteins in E. coli DH5alpha and BL21(DE3) respectively. The fusion proteins showed as different bands of different molecular weight respectively when analyzed with SDS-PAGE. Western blot demonstrated that the recombinant proteins were recognized by the positive serum of Ot. patients.

CONCLUSIONThe sta56 gene of Orientia tsutsugamushi Karp strain could be highly expressed in E. coli and its expression showed better efficiency in pET30a than in pPROEX HTb. The recombinant sta56 antigen with immunoreactivity could be used as diagnostic reagent for Ot. infection.

Animals ; Antibodies, Bacterial ; biosynthesis ; Antigens, Bacterial ; biosynthesis ; genetics ; immunology ; Bacterial Proteins ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Membrane Proteins ; biosynthesis ; genetics ; immunology ; Orientia tsutsugamushi ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Scrub Typhus ; immunology ; prevention & control

The aim of this study is to investigate the growth and proliferation of bone marrow mesenchymal stem cells (BMSCs) three-dimensionally cultured in Pluronic F-127 gel, in order to explore the cellular compatibility of gel and to investigate the feasibility of BMSCs differentiating into adipocytes in gel. Rat BMSCs were isolated from adult bone marrow, and then cultured and amplified in vitro. The BMSCs derived from the 4th passage were seeded on the scaffolds and incubated in adipogenic stimuli culture to differentiate into adipocytes. BMSCs were dispersed into gel and cultured in vitro for two weeks then the status of adhesion, growth and proliferation of the cells were observed. The edipogenic differentiation of the BMSCs was assessed by cellular morphology and further confirmed by Oil Red O staining. BMSCs were able to attach, grow and proliferate well in Pluronic F-127 gel. The BMSCs differentiated into adipocytes in gel in the presence of adipogenic stimuli over a period of 2 weeks. After only 4 days of adipogenic induction, small lipid droplets were observed within BMSCs in gel wells treated with differentiation media. At the end of 14 days, in the presence of differentiation media in gel, the size of the lipid droplets increased to occupy most of the cytoplasm, consistent with differentiation of BMSCs into adipocytes. Lipid droplets in differentiating BMSCs were positively stained with Oil Red O in the presence of differentiation media in the Pluronic F-127 treatment. We demostrated BMSCs incubated in the 3D Pluronic F-127 gel scaffolds could be induced and differentiated into adipocytes. The system for inducing differentiation of BMSCs into adipocytes is promising to apply in the construction of tissue engineering adipose tissue and the repair of fat injury, and Pluronic F-127 gel may be a suitable scaffold for cellular therapy of BMSCs.
Adipocytes ; cytology ; Adipose Tissue ; cytology ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Hydrogels ; chemistry ; Mesenchymal Stromal Cells ; cytology ; Poloxamer ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering

A 3-year-old boy had abnormal liver function, which was found in physical examination, for 5 months before admission. He had no symptoms such as anorexia, poor appetite, and jaundice, had normal growth and development, and showed no hepatosplenomegaly. Laboratory examination revealed significantly reduced ceruloplasmin (35 mg/L), as well as negative hepatotropic virus, cytomegalovirus, and Epstein-Barr virus. There were normal muscle enzymes, blood glucose, and blood ammonia and negative liver-specific autoantibodies. The boy had negative K-F ring and normal 24-hour urine copper (0.56 μmol/L). The ATP7B gene testing for the boy, his sister, and their parents detected two novel missense mutations in the boy and his sister, i.e., compound heterozygous mutations in exon 7 (c.2075T>C, p.L692P) and exon 13 (c.3044T>C, p.L1015P), which were inherited from their father and mother, respectively. Wilson's disease was confirmed by genetic diagnosis in the boy and his sister. The boy and his sister were given a low-copper diet. The boy was administered with penicillamine for decoppering and zinc supplement against copper uptake. His sister received zinc supplement alone because no clinical symptoms were observed. The boy showed normal liver function in the reexamination after 3 months of treatment.
Adenosine Triphosphatases ; genetics ; Alanine Transaminase ; blood ; Cation Transport Proteins ; genetics ; Child, Preschool ; Copper-transporting ATPases ; Female ; Hepatolenticular Degeneration ; genetics ; physiopathology ; Humans ; Liver ; physiopathology ; Male ; Mutation, Missense ; Physical Examination

OBJECTIVETo observe the effect of Tiam-l gene silencing on the metastasis of human colorectal carcinoma cell line SW480 in nude mice by real-time whole-body fluorescence imaging.

METHODSEnhanced green fluorescence protein (EGFP)-labeled human colorectal carcinoma cells, SW480/EGFP(+)/Tiam-1(-) and SW480/EGFP(+), were implanted into nude mice via tail vein injection or orthotopic colonal inoculation, and real-time whole-body fluorescence imaging was performed to compare the difference in tumor progression and metastasis between the two cells.

RESULTSBoth SW480/EGFP(+) and SW480/ EGFP(+)/Tiam-1(-) cells stably expressed EGFP, and Tiam1 gene expression was reduced in SW480/EGFP(+)Tiam-1(-) to 30% of the expression level in SW480/EGFP(+) cells. The growth rate of the two cell lines had no significant difference in vitro (P>0.05), but SW480/EGFP(+)/Tiam1(-) cell proliferation and metastasis were depressed obviously in comparison with SW480/EGFP(+) in vivo (P<0.05).

CONCLUSIONTiam-1 gene may play an important role in invasion and metastasis of human colorectal cancer.

Animals ; Blotting, Western ; Bone Neoplasms ; genetics ; metabolism ; secondary ; Cell Line, Tumor ; Cell Survival ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Diagnostic Imaging ; methods ; Female ; Fluorescence ; Gene Silencing ; Green Fluorescent Proteins ; chemistry ; genetics ; metabolism ; Guanine Nucleotide Exchange Factors ; genetics ; metabolism ; Immunohistochemistry ; Liver Neoplasms ; genetics ; metabolism ; secondary ; Lung Neoplasms ; genetics ; metabolism ; secondary ; Mice ; Mice, Nude ; Microscopy, Fluorescence ; Neoplasm Transplantation ; T-Lymphoma Invasion and Metastasis-inducing Protein 1 ; Transplantation, Heterologous

Objective　To observe the curative effect and sid e effect of the gastrokinetic agent mosapride citrate by RCT. Methods　 42 cases of functional dyspepsia (FD) were divided into two groups rando mly, the group of mosapride(21 cases):orally administrated mosapride, 5mg, t.i.d for 4 weeks, and the control (21 cases):orally administrated domperidone, 5mg, t.i.d for 4 weeks. Symptoms and side effects were recorded before and at d 14, d 28 after administration of the medicines according to GCP and double blind pri ciple. Gastric empting test was also carried out in randomly selected patients. Results　Mosapride and domperidone were significantly effective on alleviating symptoms of the patient with FD. In mosapride treated group the half emptying time was shortened and the 120 min remain rate was reduced. No sid e effect was found. Conclusion　These results suggest that mosa pride 5 mg t.i.d. is effective and safe on alleviating symptoms of patients with FD and improving the ga stric empting time.

Objective To investigate the effect of leptin on the tolerance of cultured rat brain astrocytes to ischemia and hypoxia.Methods The brain astrocytes isolated from neonatal SD rats,after purification and identification,were incubated in serum-and glucose-flee medium in the presence of 5%CO2+95%N2 for 90 min to induce isehemic and hypoxic injury. RT-PCR was performed to detect the expressions of the leptin receptors Ob-Ra and Ob-Rb in the cells, and colorimetry was used to measure the content of malonaldehyde(MDA) and lactate dehydrogenase(LDH) activity in the cell supematant.The expression level ofglial fibrillary acidicprotein(GFAP)in the cells was detected with fluorescence immunocytochemistry.Results Ischemic and hypoxic exposure of the cells induced obvious cell necrosis.Compared with the cells without the exposure,significantly decreased Ob-Rb expression(0.52±0.01 vs 1.32±0.01,P<0.05)and increased MDA,LDH and GFAP levels(709.68±47.16 vs 516.13±29.08,3.94±0.36 vs 1.81±0.21,and 0.122±0.016 vs 0.057±0.006,respectively,P<0.05) occurred after the exposure,whereas the expression level of Ob-Ra underwent no significant changes(3.87±0.13 vs 3.96±0.24,P>0.05). Compared with the exposed cells,the leptin-treated cells showed a significant reduction in MDA levels(3.94±0.36 vs 3.19±0.25,P<0.05) with significantly increased GFAP expression(0.057±0.006 vs 0.109±0.008, P<0.05)after the exposure, and the cells maintained basically intact cell morphology.Conclusion With neuroprotective effects against ischemic neuronal injuries,leptin canimprove the tolerance of rat brain astrocytes to ischemia and hypoxia.