Medicine, Chinese Traditional ; Translations

BACKGROUND: Chronic myeloid leukemia (CML) is characterized by the clonal expansion of hematopoietic stem cells. Without effective treat ment, individuals in the indolent, chronic phase (CP) of CML will undergo blast crisis (BC), the prognosis for which is poor. Therefore, it is important to clarify the mechanism underlying CML from a whole-genome perspec tive. OBJECTIVE: To investigate the gene expression profile of bone marrow mononuclear cells from CML with Applied Biosystems Expression Array System.DESIGN: Observation and controlled analysis.SETTING: Institute of Gene Engineering, Southern Medical University PARTICIPANTS: Samples of two cases of bone marrow (a chronic myeloid leukemia patient and a healthy person).METHODS: This experiment was conducted at the Institute of Gene Engineering, Southern Medical University from October 2004 to September 2005.The total RNAs were extracted and purified from bone marrow mononuclear cells derived from a CML patient and a healthy person. mRNAs were purified using an oligo (dT)-cellulose mRNA purification kits and labeled using reverse transcription, in vitro transcription (RT-IVT), then hybridized with microarray. Gene expression differentiation of the bone marrow mononuclear cells were examined by ABI 1700 Chemiluminescent Microarray Analyzer. Reproducibility of microarray results was assessed by comparing data sets obtained from the same sample and analyzed by two different arrays.MAIN OUTCOME MEASURES: ①Assessment of quality of total RNA and labled cRNA. ②Reproducibility of microarray. ③ Hybridization of array.④Results of semi-quantitative reverse transcription-polymerase chain reaction RESULTS: ①Using statistical data analysis tools, we identified 6 706 genes that were up- or down-regulated in CML patient compared with the healthy person. In these genes, we found that 17 genes were up-regulated while 51 genes were down-regulated among 68 genes closely related to CML. ②most differentially expressed genes in C/EBPalpha mediated path way and CD40L signaling pathway had reduced expression. ③Good repro ducibility of microarray was confirmed by analysis of correlation and detection concordance in technical replicates. The correlation coefficient of the detectable probe in technical replicates was 0.991 for the CML patient and 0.988 for the healthy person. ④The results of semi-quantitative RT-PCR experiments supported the reliability of our microarray analysis.CONCLUSION: By comparing expression patterns of CML with those of the healthy person, we identified a large number of genes that, were up- or down-regulated in CML patients. These data should provide useful information for finding candidate genes whose products might serve as molecular targets for treatment of CML patients.

AIM: To study the difference in glucose and lactate levels between brain extracellular fluid (ECF) and plasma in rabbits in the different blood glucose levels. METHODS: Using intracerebral microdialysis technology, brain ECF levels of glucose and lactate were measured in every 10 min under basal conditions and during a hyperglycemia-hypoglycemia clamp study. RESULTS: Under basal condition, brain ECF glucose levels were markedly lower than ambient plasma levels (30% of plasma), whereas ECF lactate levels were substantially higher (165% of plasma). During the hyperglycemia-hypoglycemia clamps, the relationship between plasma and ECF levels of glucose remained similar, but changes in ECF glucose lagged about 30 min. There were no substantially changes in ECF levels of lactate during dynamical study. CONCLUSION: There are striking differences in glucose and lactate levels between brain ECF and plasma. Lactate may involve in the metabolic process of central nervous system.

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

Objective To report a new method of fluorescent labeling technique in microarray studies: universal primer U2 labeling( UPL). The efficiency was compared of the UPL with that of random primer, restriction display labeling method and the reverse transcription coupled random primer spiking labeling method(RT-PSL). Methods Influenza viral RNA was labeled with both UPL and the conventional random primer labeling method as well as two other more laborious labeling methods( RD-direct and RD-incorporate), and hybridized with influenza virus oligonucleotide microarrays. The signals extracted from the microarrays were analyzed using SPSS 10.0 software. Results The fluorescent intensity, signal-to-noise ration(SNR), true positive ratio(TPR) of probes and labeling reproducibility of UPL were demonstrated to be higher than those of the Random primer approaches.Conclusion These results established that UPL is a valid new labeling protocol, which may have wide applications in the research and development of the microarray technology.

Objective To reversing methicillin-resistant Staphylococcus(MRS) to methicillin-susceptible Staphylococcus(MSS) by changing nutritional conditions and continuous transfer of culture .Methods MRS trains separating from clinical specimens were cultured in different conditions ,continuous cultural transfer ,and drug sensitive test were proceeded periodically to observe the phe-notypic and chemical reaction change of MRS .The mecA gene were detected of the original and mutant strains by polymerase chain reaction(PCR) ,then the gene sequenced and compared .Results 53 MRS strains were studied .6 strains were phenotype successful-ly converted to MSS in different cultural conditions ,among them mecA gene was undetected in 2 strains ,and down expressed in 4 strains .Conclusion The MRS strains separated from clinical specimens may revert to MSS by culture under different nutritional conditions .The mecA gene of MRS may be lost or lower expressed and the MRS and mutant strains may be different in genomics .

OBJECTIVETo investigate the feasibility of in vivo tumor detection using magnetic resonance (MR) molecular imaging with targeted magnetic nanoparticles as imaging probe.

METHODSTargeted probe was synthesized by covalently linking the recombinant human gonadotropin releasing hormone analog (the targeting portion) with the ultrasmall superparamagnetic iron oxide nanoparticles (the imaging portion). The imaging portion served as the control material. The in vitro tumor cell experiment and the in vivo experiment using nude mice bearing tumors were carried out to test the targeting ability of the probe. In the in vitro experiment, the targeting probe and control materials were incubated separately with A549 cells which had high affinity to gonadotropin releasing hormone. Then the cells were taken out and lysed. The resultant solution was then subjected to MR imaging. The T2 value of the solutions was measured and compared. In the in vivo experiment, the targeting probe was administered into nude mice bearing A549 tumors. Dynamic MR imaging was carried out to measure the signal and T2 value of the tumor. The control material was also administered into control group of nude mice, and dynamic magnetic resonance imaging was performed. The T2 value of the tumor in both groups were recorded and compared.

RESULTSBoth the in vitro and in vivo experiments proved the targeting ability of targeted probe. Compared with control material, the targeting probe had higher combining ability with tumor cells.

CONCLUSIONMR molecular imaging of tumor can be realized by using targeting magnetic nanoparticles.

Adenocarcinoma ; diagnosis ; pathology ; Animals ; Cell Line, Tumor ; Dextrans ; metabolism ; Drug Delivery Systems ; Feasibility Studies ; Female ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; metabolism ; Humans ; Image Enhancement ; methods ; Lung Neoplasms ; diagnosis ; pathology ; Magnetic Resonance Imaging ; methods ; Magnetics ; Magnetite Nanoparticles ; Male ; Mice ; Mice, Nude ; Molecular Imaging ; Nanoparticles ; Neoplasm Transplantation ; Recombinant Proteins ; metabolism

Objective. To ascertain the relationship between the Ala54Thr variation of FABP2 gene and general as well as regional adipose tissue depots.Subjects. 165 subjects, in which 86 were subjects with normal glucose tolerance (NGT) [age 54.45±9.80, male/female 1.05,body mass index (BMI)26.48±4.01] and 79 were subjects with non-insulin-dependent diabetes mellitus (NIDDM)(age 55.86±10.00,male/female 1.08,BMI 26.75±3.30).Design and measurements. An association study of FABP2-Ala54Thr variation detected by PCR/HhaI digestion with general and regional adipose tissue depots determined by BMI and magnetic resonance imaging [abdominal subcutaneous and visceral adipose tissue area (SA and VA) and femoral subcutaneous adipose tissue area (FA)].Results. The geneotype and allele frequencies of FABP2-Ala54Thr variation in Chinese were quite close to the frequencies in American Caucasians and Pima Indians reported in the literature. Significant difference in genotype frequency distribution was observed between FA subgroups comparisons (FA≥75cm2 versus FA＜75cm2)in NIDDM subjects (X2＝11.460,P＝0.003),with significantly increased in Thr54-carrier[Thr54(+)]genotype frequency and Thr54 allele frequency in NIDDM subject with FA＜75cm2(odd ratio for genotype was 4.62,X2＝10.112,P＝0.001;and for allele＝2.36,X2＝5.379,P＝0.020).The FA in NIDDM-Thr54(+)subgroup was significantly lower than that in subjects with NIDDM-Thr54(-)sugroup(61.19±21.51cm2 versus 75.36±31.70cm2,P＝0.021). Stepwise regression analysis revealed that FABP2-Thr54 genotype variation was an independent factor contributing to the variation of FA in NIDDM(P＝0.003).Conclusion. FABP2 is associated with regional adipose tissue depot.The decreased femoral subcutaneous adipose tissue depot in NIDDM subjects is related to FABP2-Thr54 variant.

OBJECTIVETo identify mutations of keratin 9 (KRT9) gene in a big Chinese family with epidermolytic palmoplantar keratoderma(EPPK) combined with knuckle-pad-like lesions and nail lesions.

METHODSGenomic DNA from peripheral blood of all available members in this family and 50 unrelated healthy individuals was used for amplification of the whole coding sequence and the intron-exon boundaries of KRT9 gene by PCR; The mutation was detected by direct sequence analysis and identified by restriction endonuclease Dde I.

RESULTSA mutation of AAT>AGT at codon 160 (N160S) was found in all patients but not in unaffected family members and 50 controls.

CONCLUSIONThe mutation of AAT>AGT at codon 160 (N160S) is the disease-causing mutation in this Chinese pedigree with EPPK.

Asian Continental Ancestry Group ; Codon ; DNA ; genetics ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Keratins ; genetics ; Keratoderma, Palmoplantar ; genetics ; Male ; Pedigree ; Point Mutation ; Polymerase Chain Reaction

Objective To evaluate the short-term efficacy and safety of sevelamer hydrochloride in treating maintenance hemodialysis (MHD) patients with hyperphosphemia.Methods A multicenter,open-labeled,self-control study was performed.Phosphate binders were discontinued during a two-week washout period.Patients with more than 1.78 mmol/L serum phosphorus after two-week washout period were eligible for the trial.The dose was adjusted every two weeks as necessary to achieve serum phosphorus control. Sevelamer hydrochloride was administered to 138 MHD patients for 10 weeks and a second two-week washout period followed.Results A total of 111 from 138 patients fulfilled the whole 14-week study. Mean serum phosphorus and calcium-phosphate products starte to decline after two-week sevelamer hydrochloride treatment. By the end of 10-week sevelamer hydrochloride treatment, mean serum level of phosphorus [(1.85±0.50) vs (2.57±0.54) mmol/L,P＜0.01],calcium-phosphate product [(4.16± 1.72) vs (5.79 ± 1.50) mmol2/L2,P＜0.01 ] and low density lipoprotein [(1.64±0.76) vs (2.31 ±0.87) mmol/L,P＜0.01] were significantly decreased,while the adjusted serum level of calcium and serum intact parathyroid hormone kept steady.Both serum phosphorus and calcium-phosphrus product increased after the second washout period, but the levels were still lower as compared to pre-treatment [(2.26±0.71) vs (2.57±0.54) mmol/L; (5.12±1.63) vs (5.79±1.50) mmol2/L2,P＜0.01].Of the 138 patients involved,214 episodes in 106 patients and 121 episodes in 89 patients were reported as adverse events and adverse drug reaction respectively. Gastrointestinal symptoms,of which most were mild or moderate,happened to 68.1％ (94/138) patients. Conclusions Sevelamer hydrochloride can control serum phosphorus and reduce the levels of calcium-phosphorus product and cholesterol.Slight gastrointestinal symptoms like constipation are common during the treatment.