?AIM:To compare the coherence and difference on the fundus examination made with two kinds of optical coherence tomography ( OCT): Angio-OCT and fourier domain-optical coherence tomography ( FD-OCT) .?METHODS:Using Angio-OCT and FD-OCT to measure the retinal nerve fiber layer ( RNFL ) thickness, optic parameters, and ganglion cell complexes ( GCC ) thickness from 20 subjects respectively.The coherence was tested with Pearson's correlation coefficient, the difference was tested with paired Student t testing.?RESULTS:The total correlation of the RNFL thickness, optic parameters, GCC thickness made with two kinds of OCT was between 0.7-0.8;the RNFL thickness, optic disk area etc.made with the Angio-OCT were lower than those made with FD-OCT except for the GCC thickness.?CONCLUSION: The results made with two kinds of OCT from the same subject has certain coherence, but cannot be compared directly.

To discuss clinicopathological features and molecular genetic change of congenital mesoblastic nephroma (CMN).Methods Nine cases diagnosed as CMN were analyzed retrospectively in this study.Histological features,immunohistochemical profiles and ETV6 gene rearrangement status were assessed.Results All patients were within two years of age and eight of them were within one year.The average diameter of tumors was 9.5 cm (3.2-15.0 cm).These series cases included 3 classic CMN,5 cellular CMN and 1 mixed CMN.Cystic degeneration was found in 5 cases,and cartilage islands were observed in 2 cases.Compared with classic CMN,tumor size was bigger,and hemorrhage,necrosis and mitotic figures were easily to see in cellular CMN.All the tumor cells were positive for vimentin and negative for WT-1 by immunohistochemistry.ETV6 gene rearrangement was detected in 5 cases (including 4 cellular CMN and 1 classic CMN).Three cellular CMN harbored ETV6 gene translocation,1 mixed CMN and 1 cellular CMN were negative for ETV6 gene translocation by FISH analysis.The follow up data were obtained in 7 cases and 2 cases were lost.All the 7 patients were alive without evidence of recurrence and metastasis from 5 to 46 months.Conclusion CMN is a rare infant renal tumor with unique clinicopathological characteristics.Most of cellular CMNs harbor ETV6 gene translocation.The prognosis of CMN is relative good and needs to be differentiated from other malignant renal tumors.

This study was purposed to explore the diagnostic role of flow cytometry in immunorelated pancytopenia (IRP). After 50 IRP patients were hospitalized, the concentration of serum ferritin, folic acid and vitamin B(12), immunologic test, platelet antibody, test of hepatitis A, B and C, haemolysis test and bone marrow smear examination were carried out, meanwhile the chromosome karyotype analysis and some routine examinations were performed. The 50 patients were divided into group A and group B. Group A consisted of 22 patients who were undefinedly diagnosed and intended to diagnosed as IRP, group B consisted of 28 definedly diagnosed patients with hematologic malignancies, including 7 cases of aplastic anemia, 2 of paroxysmal nocturnal hemoglobinuria, 10 of myelodysplastic syndrome, 9 of megaloblastic anemia. In addition, 30 normal people were used as normal control group (group C). For groups A and B, the binding autoantibodies of bone marrow stem/progenitor cells, erythroblasts and myelocytes were detected by flow cytometry, meantime the ratio of total B-(CD10(+)) and CD5(+) B-lymphocytes in peripheral blood was assayed. For control group, the ratios of CD19(+) and CD5(+) B lymphocytes in peripheral blood were determined alone. The results indicated that the detection of bone marrow autoantibodies in 20 patients of group A showed positive with 90.90%. The IgG type was found mostly in antibody binding types, next the IgM type, the IgA type was fewer. The detection of bone marrow autoantibodies of 2 patients in group B showed positive with 7.14%. The positive rate in group A was obviously higher than that in group B (p < 0.01). The ratios of CD19(+) and CD5(+) B lymphocyte in peripheral blood were significant higher in group A than that in group B and control group (p < 0.01), but there was no significant difference between groups B and control. It is concluded that the application of flow cytometry in detecting the autoantibodies of bone marrow cells and CD19(+) B-and CD5(+) B-lymphocyte in peripheral blood can provide reliable diagnostic evidence and detection measure for diagnosis and differential diagnosis of IRP, as well as may contribute to draw up more effective therapeutic strategy.
Adult ; Aged ; Autoantibodies ; immunology ; B-Lymphocytes ; immunology ; Case-Control Studies ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; diagnosis ; immunology ; Young Adult

In order to evaluate the immunogenicity of HPV 58 L1 DNA vaccines, five DNA vaccines had been constructed with pcDNA3.1 vector containing different L1 genes of HPV 58, which were designated as L1h, L1hDeltac, L1S, L1SM and L1wt. The protein expression of DNA vaccines in vitro was tested by Western blot. The ability of forming pseudovirus was evaluated by transfecting DNA vaccine together with pcDNA3.1-h58L2 and pcDNA3.1-GFP into 293FT cells. The neutralizing antibodies and cellular immune response produced in BALB/c mice immunized with the DNA vaccines were detected by using pseudovirus-based neutralization assay and ELISPOT respectively. The results showed that the five DNA vaccines had been successfully constructed; the level of protein expression of L1hDeltac was the highest and those for L1S and L1SM were of medium, while no expressed target protein of L1wt was detected. Only L1S could form the pseudovirus while the other four vaccines could not. L1S and L1h could induce neutralizing antibody. However, the average titer of neutralizing antibody for L1S (1:6,400) was much higher than that for L1h (1:48) and the other three vaccines could not induce neutralizing antibody. No cellular immune response for all five DNA vaccines was detectable by ELISPOT. The results indicated that DNA vaccine against HPV 58 can form pseudovirus in vitro, also can induce high level of neutralizing antibodies. This provides reference for screening HPV vaccine in future.
Animals ; Blotting, Western ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Female ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Models, Genetic ; Neutralization Tests ; Papillomaviridae ; immunology ; Papillomavirus Vaccines ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vaccines, DNA ; genetics ; immunology

After the preparation of cationic liposomes composed of DDAB/DOPE, cationic liposome-DNA complexes and lipid-polycation-DNA (LPD) complexes were formulated, respectively. Gel retardation assay was employed to select appropriate ratios of cationic liposomes to DNA of the liposome-DNA complexes. The morphology of LPD and liposome-DNA complexes was observed by transmission electron microscopy. The diameter and surface charge of LPD and liposome-DNA complexes were measured by photon correlation spectroscopy (PCS). Their transfection efficiencies in Chang cells and HepG2 cells were evaluated by beta-gal assay kit. It was found that LPD and liposome-DNA complexes had a regular spherical surface. However, compared with liposome-DNA complexes, LPD had rather smaller particle size and much higher transfection efficiency in Chang cells and HepG2 cells in vitro. LPD could be prepared easily with small particle sizes and high transfection activities. LPD may be a good non-viral gene delivery vehicle for applications in gene delivery.
Cations ; DNA ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Lipids ; chemistry ; Liposomes ; chemistry ; Liver Neoplasms ; metabolism ; pathology ; Protamines ; chemistry ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo investigate the prognostic value of a modified WPSS based on routine laboratory parameters in Chinese patients with myelodysplastic syndromes (MDS).

METHODSOne hundred and sixty four adult MDS patients were retrospectively analyzed.

RESULTSThe median follow-up time was 19 (1-138) months and the median survival (MS) was 36 months. 2-year prospective survival (PS) was 60% and 5-year PS was 42%. In patients with very low-risk, low-risk, intermediate-risk, high-risk and very high-risk stratified by WPSS, 2-year PS was 100%, 96%, 81%, 38% and 14%, and 5-year PS was 100%, 83%, 54%, 20% and 0, respectively (P<0.01). Among parameters of laboratory routine examination, elevated mean cell volume (MCV), mean cell hemoglobin (MCH), neutrophil alkaline phosphatase (N-ALP) index and nucleated RBC PAS negative were good prognostic factors, while reduced Hb, BPC and bone marrow elevated blasts, dysplasia more than 1 lineage, and lymphocyte-like micromegakaryocyte (MEGly) as well as elevated serum LDH were poor prognostic factors in uni-variable analysis. Among them, MCV, MEGly and blast had independent prognostic significance in multi-variable analysis (P = 0.011, 0.013 and 0.016, respectively). WPSS was modified by omitting chromosomal karyotype and transfusion dependence and adding MCV and MEGly. In patients with low-risk, intermediate-risk and high-risk stratified by modified WPSS, 2-year PS was 94%, 68% and 49%, respectively; and 5-year PS was 86%, 53% and 14%, respectively (P<0.01).

CONCLUSIONThe modified WPSS worked well for prognostic prediction in Chinese patients with MDS.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo apply the WHO criteria and the minimal diagnostic criteria to the classification of myelodysplastic syndromes (MDS) with low percentage (< 0.050) bone marrow (BM) blasts.

METHODSTwo hundred and ten MDS patients with less than 0.050 BM blasts diagnosed between 1988 and 2005 according to FAB criteria were retrospectively reclassified with WHO criteria (2001) and minimal diagnostic criteria.

RESULTSAccording to the WHO criteria, 5 patients were diagnosed as refractory anemia (RA), 7 as refractory anemia with ringed sideroblasts (RARS), 76 as refractory cytopenia with multilineage dysplasia (RCMD), 9 as RCMD-RS, 35 as MDS-unclassified (MDS-U), 3 as 5q - syndromes, and the rest 75 patients could not be classified suitably. Among the latter 75 patients 16 BM smears showed dysplasia in more than 2 cell lineage but only unilineage cytopenia in peripheral blood (PB). Nine of them were reclassified as RCMD after followed up for more than half a year. Forty-four BM smears showed erythroid dysplasia only, but bicytopenia or pancytopenia in PB. Twenty-seven of them were classified as RCMD after follow-up. Fifteen BM smears not showed dysplasia in any myeloid lineage were reclassified as MDS (5 patients), HS-MDS (5 patients) and idiopathic cytopenia of uncertain significance (ICUS) (5 patients) according to the MDS minimal diagnostic criteria.

CONCLUSIONAccording to WHO criteria (2001), RA is the least diagnosis in MDS. The minimal diagnostic criteria for MDS classification of patients not fulfilled the standard criteria of MDS.

Adolescent ; Adult ; Aged ; Anemia, Refractory ; etiology ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; classification ; complications ; diagnosis ; pathology ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo explore the value of routine laboratory parameters in diagnosis of myelodysplastic syndromes (MDS) and differential diagnosis of patients with hypoplastic MDS from chronic aplastic anemia (CAA) for providing reference standard for primary hospitals.

METHODSThe laboratory parameters at diagnosis of 152 MDS patients with less than 0.05 bone marrow blasts and 86 CAA patients were retrospectively analyzed.

RESULTSThere were significant differences between MDS and CAA in Hb, red cell distribution width-coefficient variation (RDW-CV), immature reticulocyte fraction (IRF), BPC, the ratio of G1 (the sum percentage of myeloblast and promyelocyte) to G2 (the sum percentage of neutrophilic myelocyte and metamyelocyte) (Ratio G), the ratio of El (the sum percentage of proerythroblast and early erythroblast) to E2 (the sum percentage of intermediate erythroblast and late erythroblast) (Ratio E), megakaryocyte count (Meg), erythroblast PAS, neutrophil alkaline phosphatase (N-ALP), and serous levels of indirect bilirubin (IBIL), lactose dehydrogenase (LDH), folic acid (FA), VitB12 and ferritin. Chromosome abnormalities were found in 74 MDS patients (48.7%) but in none of CAA patients (P < 0.001). Furthermore, for differentiating MDS with less than 0.05 blasts from CAA, the sensitivity and specificity of combination of Meg, PAS, and IBIL level was 89.1% and 92.7%, the Youden index (gamma) was 0.818. Moreover, in the seven hypoplastic MDS cases, BPC, myeloblast percentage, Ratio G, Meg, erythroblast PAS and FA were statistically different from those of CAA; the sensitivity and specificity of combination of PAS and BPC was 85.7% and 100%, the gamma was 0.857; the sensitivity and specificity combination of Ratio G, Meg PAS was 85.7% and 98.8% respectively, the gamma was 0.845.

CONCLUSIONThe routine laboratory parameters, especially BPC, Meg, Ratio G, PAS, IBIL may be helpful for the diagnosis of MDS and differential diagnosis of hypoplastic MDS from CAA.

Adolescent ; Adult ; Aged ; Anemia, Aplastic ; diagnosis ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; Retrospective Studies ; Young Adult

OBJECTIVETo assess the clinical diagnostic value of 18F-FDG imaging by coincidence circuit SPECT with low-dose CT in differential diagnosis of pulmonary lesions and mediastinal lymph node involvement, which can not be definitely diagnosed based on regular CT image in patients with non-small-cell lung cancer (NSCLC).

METHODSBy using GE-Millennium VG with Hawkeye, 18F-FDG imaging was carried out in 48 patients with suspected lung cancer. Clinical value of 18F-FDG imaging for diagnosing malignancy was evaluated through comparison with the final pathological results. Mediastinal lymph node involvement was also assessed through lesion-by-lesion comparison with pathologic results in 74 lymph node regions from 24 patients.

RESULTSFinal pathologic diagnoses of these patients were 36 malignancies consisting of 20 adenocarcinomas, 12 squamous cell carcinomas, 3 small cell carcinomas and I large cell carcinoma; 12 benign tumors including 6 pneumonias, 2 tuberculosis, 2 hamatomas, 1 cyst and 1 neurofibroma. Of 48 patients, uptake of 18F-FDG in the chest was found to be abnormal in 40. Correct diagnosis were made in 34 malignancies and 6 false positive lesions were excluded based on morphology and 18F-FDG uptake status of the lesion. There were 6 false positive and 2 false negative cases. Furthermore, extrathoracic metastases which were not showed on previous CT image in 4 patients including one in the adrenal gland and 3 in the bone were detected by 18F-FDG imaging. The sensitivity, specificity and accuracy of the 18F-FDG imaging for differentiating malignant tumor from benign was 94.4%, 50.0% and 83.3%, respectively. Squamous cell carcinoma was found to uptake more FDG than adenocarcinoma. For determination of mediastinal lymph node involvement, the sensitivity, specificity and accuracy of 18F-FDG imaging was 57.9% , 90.9% and 82.4%, respectively through lesion-by-lesion comparison; whereas, which was 61.5%, 81.8% and 70.8%, respectively, based on case-by-case comparison.

CONCLUSION18F-FDG imaging by coincidence circuit SPECT with low-dose CT is quite helpful in differential diagnosis for patient with undetermined lesion on regular CT image, but it is limited for staging of lung cancer in the patients with non-small cell lung cancer.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; diagnostic imaging ; Diagnosis, Differential ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lung ; diagnostic imaging ; pathology ; Lung Neoplasms ; diagnosis ; diagnostic imaging ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonia ; diagnosis ; diagnostic imaging ; Preoperative Care ; Radiation Dosage ; Radiopharmaceuticals ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed ; Tuberculosis, Pulmonary ; diagnosis ; diagnostic imaging

Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.
Amino Acid Sequence ; Animals ; Codon ; genetics ; Gene Expression ; Pichia ; genetics ; metabolism ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Scorpion Venoms ; genetics ; isolation & purification ; metabolism ; Scorpions ; chemistry ; Transformation, Genetic