Adolescent ; Capillary Leak Syndrome ; chemically induced ; Female ; Granulocyte Colony-Stimulating Factor ; adverse effects ; Humans ; Male ; Middle Aged

Objective To investigate the diagnostic value of preoperative real-time three-dimensional color Doppler transthoracic echocardiography on assessment of patients with structural heart disease (SHD). Methods A total of 111 pa?tients were assessed preoperatively using real-time three-dimensional color Doppler transthoracic echocardiography (RT-3D-CDTTE), which include 31 SHD patients and 80 patients without SHD that were designed as negative control. Conven?tional two-dimensional color Doppler transthoracic echocardiography (2D-CDTTE) were used to compared with RT-3D-CDTTE while cardiovascular angiography and intraoperative findings were used as“Golden Standard”simultaneously. First of all, preoperative echocardiographic examination were performed and diagnosis was given. Angiography and intraoperative findings were hired to verify the accuracy of echocardiographic diagnosis before operation. Results (1) The preoperative RT-3D-CDTTE displayed three-dimensional structure and hemodynamic status of SHD cardiac lesions clearly, which were consistent with cardiovascular angiography and intraoperative findings. (2) P value of McNemar test between 2D-CDTTE and“Golden Standard”was greater than 0.05, Kappa=0.654 (P<0.001). P value of McNemar test between RT-3D-CDTTE and“Golden Standard”was greater than 0.05, Kappa=0.932 (P<0.001). Conclusion RT-3D-CDTTE can provides essen?tial information for preoperative evaluation which is important for decision of SHD management.

Objective To analyze blood Met、 Phe 、Tyr、 Arg、 Cit、 Orn、 Ser、 Thr、 C0、 C2、 C3、 C14、C14 ∶ 1 , C16 , C16 ∶ 1 , C18 , C18 ∶ 1 and urine 4-OH-PHPLA , 4-OH-PHPPA level of NICCD patient and discuss the application value of diagnosis NICCD. Methods From May 2011 to May 2015, 21 NICCD patient were diagnose in Guangxi Newborn Screening Center. Meanwhile, 100 normal children were selected as the control group. Blood Met, Phe, Tyr and other factors and urine 4-OH-PHPLA, 4-OH-PHPPA level were analyzed by SPSS. Results In the experimental group, blood Met, Phe, Tyr and many other indexes and urine 4-OH-PHPLA, 4-OH-PHPPA level were higher and blood Orn/Cit were lower than the control group(P < 0.05), while blood C2and Cit/Arg were increased (P > 0.05). Conclusion NICCD patient has abnormal biochemical index. Blood test by TMS and urine test by GC-MS are very important in NICCD diagnosis.

AIM:To investigate the expression and potential role of complement 5a receptor (C5aR) in chro-nic graft-versus-host disease (cGVHD).METHODS:The expression of C5aR on lymphocytes and the frequency of CD4+CD25+ Foxp3+ regulatory T cells (Tregs) in 20 cGVHD patients and 9 healthy donors was detected by flow cytometry.The correlation between the expression of C5aR and the percentage of Tregs in the cGVHD patients was analyzed.In addition, the splenocytes from the mice were cultured in vitro, and stimulated these splenocytes with recombinant mouse C5a protein (rmC5a).The peripheral blood mononuclear cells (PBMCs) from cGVHD patients were cultured in vitro, which was inhibited by C5aR antagonist (C5aRA).The frequency of Tregs in these splenocytes and the PBMCs were evaluated by flow cytometry.RESULTS:The expression of C5aR on the lymphocytes was significantly increased in the cGVHD patients compared with the healthy donors, while the percentage of Tregs was markedly lower in the cGVHD patients.The expression of C5aR was negatively correlated with the percentage of Tregs.Furthermore, the development of Tregs was suppressed by rmC5a stimulation, but was promoted by C5aRA in vitro.CONCLUSION:C5aR elevation is associated with Treg reduction in cGVHD, indicating that C5aR may play a potential role in suppressing Tregs, resulting in the incidence of cGVHD.

BACKGROUNDIn general, it is very important to understand the state of T cell immune response against tumor cells in leukemia patients and it is especially critical to assess the T cell repertoire of untreated patients. As we know, few studies have dealt with the distribution of oligoclonal T cells in leukemia, so we investigated the distribution and clonality of TCR Vbeta repertoire of T cells in patients with chronic myelogenous leukemia (CML) in chronic phase.

METHODSThe complementarity determining region 3 (CDR3) of TCR Vbeta24 subfamily genes were amplified in peripheral blood mononuclear cells from 27 cases with CML using reverse transcription-polymerase chain reaction (RT-PCR). In order to observe the distribution of TCR Vbeta repertoire, the PCR products were further analyzed by genescan technique to evaluate clonality of the detectable TCR Vbeta T cells. The PCR products of the oligoclonal T cells from three cases were analyzed by direct sequencing to define the sequence of CDR3.

RESULTSThe expression pattern of TCR Vbeta repertoire in different individuals are different. Vbeta2-21 subfamilies could be detected in CML cases. The frequent usage Vbeta repertoire in CML was Vbeta1, Vbeta2 or Vbeta13. Most of the PCR products from 27 patients displayed polyclonality, while a part of the PCR products from 21 out of 27 samples displayed clonal expansion pattern. The clonal expanded T cells in CML could be found in Vbeta16 subfamilies. The frequent usage of Vbeta genes in clonal expansion was Vbeta3, Vbeta13 or Vbeta21. Multiple Vbeta clonal expansion was a general phenomenon in the same patient. The CDR3 sequence of Vbeta21 oligoclonal T cells from 3 cases showed some difference in splice regions and in the usage of J segments.

CONCLUSIONSThese results indicated that clonal expanded T cells could be found in patients with CML and were tendentious in Vbeta3, Vbeta13 and Vbeta21 subfamilies that may be related to the specific immune response for leukemia cell associated antigen.

Clone Cells ; Complementarity Determining Regions ; analysis ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; Receptors, Antigen, T-Cell, alpha-beta ; analysis ; T-Lymphocytes ; immunology ; pathology

OBJECTIVETo investigate the clonal expansion of T cell receptor (TCR) Vbeta subfamily T cells from cord blood induced by bcr3-abl2 peptide in vitro.

METHODST cells from 3 units of cord blood were amplified by anti-CD(3) monoclonal antibody (McAb) and IL-2 with or without synthetic b3a2 peptide. T cell specific cytotoxicity was analyzed by lactate dehydrogenase (LDH) assay, TCR Vbeta subfamilies by using reverse transcriptase-polymerase chain reaction (RT-PCR) and genescan technique.

RESULTSbcr3-abl2 peptide specific cytotoxicity T cells were successfully induced from the 3 units of cord blood by synthetic b3a2 peptide. Compared with that in CD(3) McAb induced cells, distribution pattern of TCR Vbeta repertoire was different in T cells induced with b3a2 peptide. Oligoclonal and oligoclonal tendency TCR Vbeta subfamily T cells could be identified in cord blood T cells induced by b3a2 peptide in 1 or 2 weeks, whereas those induced by anti-CD(3) McAb and IL-2 were mostly polyclonal.

CONCLUSIONThe cytotoxicity T cells with anti-CML specificity could be induced by b3a2 peptide. The specific anti-CML cytotoxicity may be derived from the clonal expansion TCR Vbeta subfamily T cells.

Antibodies, Monoclonal ; immunology ; CD3 Complex ; immunology ; Fusion Proteins, bcr-abl ; pharmacology ; Genes, T-Cell Receptor beta ; Humans ; Interleukin-2 ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo screen the molecular markers for refractory anemia with excess blasts in transformation (RAEB) in myelodysplastic syndromes (MDS) by serum proteome profiling.

METHODSThe serum protein were isolated from patients with RAEB, acute myeloid leukemia or normal subjects by 2-dimensional electrophoresis (2-DE), and the electrophoresis gels were obtained to identify the differentially reacting protein spots. The replica gels of the differentially reacting proteins were analyzed to locate the matching protein spots, which were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) and database searching.

RESULTSSeven differentially expressed proteins in RAEB were found by 2-DE. Of the 7 proteins, 4 were identified by MALDI-TOF-MS to have significantly differential expression in RAEB, including dipeptidyl peptidase (DPP/CD26), polymerase (DNA directed) kappa, PRO2044 and an albumin-like protein.

CONCLUSION2-DE-based serum proteome profiling helps identify serum proteomic biomarkers related to MDS. DDP/CD26 has increased expression in the serum in RAEB subtype MDS, suggesting its possible role in advanced MDS.

Anemia, Refractory, with Excess of Blasts ; blood ; genetics ; Bone Marrow ; pathology ; DNA-Directed DNA Polymerase ; blood ; Dipeptidyl-Peptidases and Tripeptidyl-Peptidases ; blood ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; classification ; genetics ; Proteomics

OBJECTIVETo explore the necessity of large-scale screening of mitochondria DNA (mtDNA) A1555G mutation for prevention of aminoglycoside antibiotic induced deafness in newborns.

METHODSOne thousand blood filter samples were collected from neonates born in July 2008 in Shenzhen. DNA was extracted with Chelex-100 Resin and amplified by PCR. The mtDNA A1555G mutation was determined by denaturing high-performance liquid chromatography(DHPLC) for PCR products. The positive frequency was calculated.

RESULTSThe mitochondrial DNA A1555G mutation was detected in 2 cases of 1000 neonates. The frequency of mutation was 0.2%.

CONCLUSIONThere is a high frequency of mtDNA A1555G mutation in neonates, the large-scale screening of mtDNAA1555G mutation in newborns might detect the individuals sensitive to aminoglycoside antibiotic, which is helpful to guide a rational medication for newborns and the maternal relatives at high-risk. Furthermore, it might be useful to prevent aminoglycoside antibiotic induced deafness.

Base Sequence ; DNA Mutational Analysis ; methods ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Infant, Newborn ; Male ; Polymerase Chain Reaction

OBJECTIVETo evaluate the economic significance of Meek skin grafting and automicrografting combined with large piece of allogenous skin (micrografting in brief) in the treatment of patients with extensive deep burn.

METHODSTwenty-four patients with extensive deep burn admitted to the First Affiliated Hospital of Wenzhou Medical College were divided into Meek skin grafting group and micrografting group, with 12 patients in each group. Statistical comparison between Meek skin grafting group and micrografting group in respect of wound healing time, consumption of each special dressing, total cost of hospitalization, rehabilitation cost during convalescence was made. Then the cost and effect value was compared between two groups.

RESULTSThe wound healing time, consumption of each special dressing, total cost of hospitalization and rehabilitation cost in Meek skin grafting group was (14.4 +/- 1.9) d, yen(16 590 +/- 521), yen(421 628 +/- 145), yen(39 571 +/- 225), respectively, and that in micrografting group was (25.6 +/- 4.2) d, yen (136 441 +/- 356), yen(539 526 +/- 686), yen(55 853 +/- 794), respectively. The difference between two groups were statistically significant (P < 0.01).

CONCLUSIONSIn a definite range of burn size, Meek skin grafting has a lower therapeutic cost and better therapeutic effects as compared with micrografting.

Adolescent ; Adult ; Burns ; surgery ; Female ; Humans ; Male ; Middle Aged ; Skin Transplantation ; economics ; methods ; Surgical Flaps ; Young Adult

The rearrangement segments of TCR Valpha40 gene with Jdelta1, Ddelta3 or psi Jalpha were amplified in genomic DNA from peripheral blood mononuclear cells of 10 normal subjects, sorted CD3(+) cells from peripheral blood of 4 cases and thymocytes from 7 cases, by using nested PCR. Different amounts of DNA from all samples were amplified to estimate the frequency of Valpha40 gene rearrangements. The results indicated that the rearrangements of TCR Valpha40 gene with Jdelta1, Ddelta3 or psi Jalpha could be found respectively in the most samples of peripheral blood T cells or thymocytes. The frequencies of Valpha40 rearrangements were different in peripheral blood T cells and thymocytes by analysis of PCR with different amounts DNA. It is concluded that the TCR V alpha40-psi Jalpha was the most frequent rearrangement in mature and immature T cells, whereas TCR Valpha40-Ddelta3 was more frequently rearranged in immature T cells
Gene Rearrangement ; Humans ; Polymerase Chain Reaction ; Protein Subunits ; genetics ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; T-Lymphocytes ; physiology