Background:The clinical outcome of transtrochanteric rotational osteotomy (TRO) for osteonecrosis of the femoral head (ONFH) remains controversial,and the promising clinical results of several Japanese studies could not be reproduced in American and European studies.Trying to solve controversies on TRO for ONFH rising from apparently conflicting studies,a meta-analysis was conducted to assess the 5-and 10-year hip survival rates (with conversion to artificial joint replacement and radiographic failure as endpoints) after TRO.Methods:All eligible studies were searched in seven comprehensive databases including PubMed,Web of Science,Embase,Cochrane Library,VIP Database,China Knowledge Resource Integrated Database,and Wan Fang Database prior to June 2019.The outcomes evaluated were 5-and 10-year hip survival rates after TRO.The odds ratio and risk difference for the non-comparative binary data with the 95％ confidence intervals (CIs) were calculated for each outcome.The included studies were assessed for methodologic bias and potential reasons for heterogeneity were explored.Results:Nineteen studies of TRO for ONFH were eligible for this meta-analysis according to inclusion criteria.Based on the previous report,two calculation methods (Methods 1 and 2) were adopted in this meta-analysis.Furthermore,we performed a subgroup analysis of the 5-and 10-year hip survival rates (Method 1) after TRO for ONFH:Asian sub-population and non-Asian subpopulation.Taking conversion to artificial joint replacement as the endpoint,5-and 10-year hip survival rates (Method 1) after TRO for ONFH in the Asian population were 0.86 (95％ CI =0.82-0.89) and 0.72 (95％ CI =0.65-0.78),respectively,and 5-and 10-year hip survival rates after TRO for ONFH in the non-Asian population were 0.55 (95％ CI =0.43-0.67) and 0.42 (95％ CI =0.28-0.55),respectively.The 5-and 10-year hip survival rates (Method 2) after TRO for ONFH were 0.90 (95％ CI =0.79-0.95) and 0.89 (95％ CI =0.81-0.94),respectively.Taking radiographic failure as the endpoint,5-and 10-year hip survival rates after TRO for ONFH were 0.70 (95％ CI =0.64-0.76) and 0.53 (95％ CI =0.46-0.61),respectively.Conclusions:The 5-and 10-year hip survival rates after TRO for ONFH were satisfactory in the Asian population,and were acceptable in the non-Asian population despite high early failure rates.

Humans ; Siblings ; Forensic Medicine ; Forensic Genetics

To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

This study was purposed to characterize the first case of acute promyelocitic leukemia (AML-M(3a)) with t(15;17), trisomy 8 and tetrasomy 8, and explore its characteristics of morphology, cytogenetics, molecular biology, immunology and clinical features. Morphological changes of peripheral blood and bone marrow smears were observed under microscope. Chromosome specimen was prepared by 24 h short-term culture of bone marrow cell, RHG-banding technique was used for karyotypic analysis. PML-RARa fusion gene transcript was detected by nested-reverse transcription-polymerase chain reaction (nested RT-PCR). Interphase fluorescence in situ hybridization (FISH) using chromosome 8 centromere specific probe were carried out to detect abnormal numbers of chromosome 8. Immunophenotypic analysis was performed by flow cytometry. The results showed that peripheral blood smear revealed 65% promyelocyte, and bone marrow aspirate was hypercellular with 72.4% promyelocyte, moderately basophilic cytoplasm with numerous azurophilic granules. Karyotype analysis demonstrated 48, XY, +8, +8, t(15;17)(q22;q12) [16]/47, XY, +8, t(15;17)(q22;q12) [3]/46, XY, t(15;17)(q22;q12) [1]. RT-PCR assay revealed PML-RARa fusion gene transcript (+). FISH showed that the percentages of cells exhibiting 1, 2, 3, 4, 5, 6 green fluorescence signals were 0.5, 7, 19, 55, 18 and 0.5, respectively. This confirmed the presence of tetrasomy 8 and trisomy 8 and also revealed a low percentage of a pentasomy 8 clone. Immunophenotypes of the blasts displayed that CD13 (96.2%), CD33 (55.9%), CYMPO (93.5%) were positive. All the lymphoid markers tested were negative. The patient survival time was just 10 days. It is concluded that tetrasomy 8 is secondary cytogenetic event after t(15;17) in this case. It may be a consequence of clonal evolution of trisomy 8. t(15;17) AML-M(3) with tetrasomy 8 heralds a poor prognosis.
Chromosomes, Human, Pair 15 ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, Pair 8 ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Promyelocytic, Acute ; genetics ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; Oncogene Proteins, Fusion ; genetics ; RNA, Messenger ; analysis ; Translocation, Genetic ; Trisomy

AIMTo convenience of the methods for assaying red blood cell glycolysis without oxygen condition in the studies.

METHODSReagent kit of glucose, perchloric acid, visible light prismatic photometer, battle of nitrogen and rocking bed are used in the studies. The process includes 4 steps prepare Tris- HCI solution and so on, assay of red blood cell glycolysis without oxygen condition and account of glycolysis rate.

RESULTSHuman red blood cells stored at 4 degrees C for 75 d, in SOD solution, the glycolysis rate is 86.2% +/- 5.0%, distinctly better than GMA solution (39.2% +/- 8.9%).

CONCLUSIONThe methods of assaying glycolysis without oxygen condition not use Habea's apparatus. The operation is convenient and simple and its determinations can be performed in ordinary laboratory and is is accurate.

Erythrocytes ; metabolism ; physiology ; Glycolysis ; physiology ; Hematologic Tests ; methods ; Humans ; Oxygen ; metabolism

AIMTo establish a simple and effective method for RBCs labeling and survival assays, and the qualities of rabbit RBCs preserved in GMA solution at 4 degrees C were verified.

METHODSThe bloods were taken through the ear arteries of the rabbits. The RBCs were labeled by fluorescein isothiocyanate (FITC), and were reinjected to the same rabbit through ear veins. The percentage of FITC labeled RBCs was assayed by FACS at a series of times after injection. The SAS software was employed to analyze the data and establish the regression equations. The 24-hour recovery and the half-life span of the labeled RBCs were calculated according to the equations.

RESULTSThe 24-hour recovery and the half-life span of the labeled RBCs in the control group were 93.76% +/- 5.40% and 22.50% +/- 4.37 days respectively, which was in agreement with the previous papers. The 24-hour recovery and the half-life span of the labeled RBCs in the GMA group were 89.13% +/- 7.10% and 11.41% +/- 1.63 days respectively, which was coincident with the infusion conditions.

CONCLUSIONCompared with other methods of RBCs labeling in vivo, FITC labeling was thought to be easier and cheaper to use, which could facilitate the analysis of the biological character of the labeled cells, and could be used to trace the fate of labeled cells.

Animals ; Blood Preservation ; methods ; Erythrocyte Aging ; physiology ; Erythrocyte Count ; Erythrocytes ; physiology ; Fluorescein-5-isothiocyanate ; Rabbits ; Software

OBJECTIVETo investigate pathological changes in the epileptogenic foci of children with intractable epilepsy and their clinical significance.

METHODSThirty children with intractable epilepsy were included in the study. The epileptogenic foci were surgically resected and pathological changes in the obtained specimens were observed under a light microscope (LM) and a transmission electron microscope (TEM).

RESULTSUnder the LM, cortical dysplasia was found in 14 cases (47%), hippocampal sclerosis in 11 cases (37%), dysembryoplastic neuroepithelial tumor in 1 case (3%), ganglioglioma in 1 case (3%), and encephalomalacia in 3 cases (10%). The TEM observation revealed pathological changes in the ultrastructure of the hippocampus and extra-hippocampal cortex, such as changes in the number of synapses and synaptic structure, decrease in neurons and karyopyknosis, swelling and degeneration of astrocytes, and changes in mitochondrial structures.

CONCLUSIONSPathological changes in the hippocampus and extra-hippocampal cortex, especially synaptic remodeling, may be the morphological basis for spontaneous recurrent seizures in children with intractable epilepsy. The pathological changes and epileptiform activity are related to an imbalance between excitatory and inhibitory neurotransmission.

Adolescent ; Brain ; pathology ; ultrastructure ; Cerebral Cortex ; pathology ; ultrastructure ; Child ; Child, Preschool ; Epilepsy ; pathology ; surgery ; Female ; Hippocampus ; pathology ; ultrastructure ; Humans ; Infant ; Intelligence ; Male ; Microscopy, Electron, Transmission

To study the dynamic changes of ultrastructure of erythrocytes in prolonged preservation of blood with preservative fluid containing superoxide dismutase (SOD), the whole blood samples were preserved at 4 degrees C in SOD-containing solution, the morphologic changes of erythrocyte were dynamically ob served by transmission microscopy after preservation for 42, 75 and 85 days, an d the blood samples preserved in GMA solution served as control. Three variance was applied to analyze the data with SAS software. The results showed that the metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were lower than those of erythrocytes preserved in GMA solution. Most of metamorphotic rates of erythrocyte preserved in SOD-containing solution for 42, 75 and 85 days were correspond to those of erythrocytes preserved in GMA solution for 42 days, or even lower. It is concluded that SOD-containing preservative fluid might help to maintain the normal morphology of erythrocytes in prolonged preservation at 4 degrees C.
Blood Preservation ; Erythrocyte Deformability ; Erythrocytes ; ultrastructure ; Humans ; Microscopy, Electron ; Superoxide Dismutase ; pharmacology ; Time Factors

OBJECTIVESTo investigate CD3+CD56+ lymphocytes and their subsets in the peripheral blood of chronic hepatitis B patients and to explore the relationship between these cells and the pathogenesis of their diseases.

METHODSBlood samples from 53 chronic hepatitis B patients, 17 from HBV asymptomatic carriers (ASC) and 19 from healthy controls (HC) were collected. CD3+CD56+ lymphocytes were detected by flow cytometry (FCM), then the CD3+CD56+ lymphocytes were gathered to analyze their expressions of CD4, CD8, TCR Valpha24, TCRalpha/beta and TCRgamma/delta.

RESULTSThe number of CD3+CD56+ lymphocytes of chronic hepatitis B patients (7.4+/-4.6%) was more than those of ASC (4.5%+/-3.5%) and healthy controls (4.4%+/-3.7%). The expressions of TCR Valpha24 on CD3+CD56+ lymphocytes showed no significant differences among the three groups, but the expression of TCR Valpha24 on CD3-CD56+ lymphocytes of ASC ( 2.8%+/-1.4% ) was much more than that of the HC (1.7%+/-1.0%). For the subsets analysis, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of chronic hepatitis B (61.9%+/-16.8% and 68.1%+/-16.9%) were significantly higher than those of the HC (49.2%+/-15.6% and 56.4%+/-17.9%), while the TCRgamma/delta subsets of chronic hepatitis B and ASC (29.6%+/-15.4% and 30.5%+/-14.8%) were decreased significantly than those of the HC (41.4%+/-19.4%). On the other hand, the CD8 and TCRalpha/beta subsets of CD3+CD56+ lymphocytes of severe chronic hepatitis B (69.0%+/-14.0% and 76.1%+/-12.9%) and CD8 subsets of moderate chronic hepatitis B patients (66.4%+/-14.9%) were significantly higher than those of the mild chronic hepatitis B patients (51.4%+/-16.2% and 62.1%+/-14.6%).

CONCLUSIONThe pathogenesis of chronic hepatitis B may positively relate to the high expression of CD8 on the CD3+CD56+ lymphocytes.

Adult ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

BACKGROUNDThis study was designed to detect methylation of E-cadherin gene promoter and gene mutation of beta-catenin in exon 3 and their expression of protein and mRNA in primary tumor and lymph node metastatic tumor of nasopharyngeal carcinoma (NPC), and investigate the mechanism of invasion and metastasis of neoplastic cells in NPC.

METHODSFourty-two fresh biopsy samples were taken from untreated NPC patients at the Affiliated Hospital of Sun Yat-sen Medical College, Sun Yat-sen University, Guangzhou, China during the period of 1999-2002. Among them 21 were taken from primary tumors and the other 21 from lymph node metastatic tumors. The gene promoter methylation of E-cadherin was detected by methylation-specific PCR (MSP). The mutation in exon 3 of beta-catenin was detected by direct sequencing analysis. RT-PCR, Western blot and immunohistochemical staining were used to detect the mRNA and protein expression patterns in both primary and metastatic tumors of NPC.

RESULTSDown-regulated expression of E-cadherin in metastatic tumor was compared with that in primary tumor. Reduced expression of E-cadherin was found to be correlated with lymph node metastatic tumor of NPC (P = 0.004); but there was no obvious correlation between primary and metastatic tumors in the expression of beta-catenin (P = 0.698). The mRNA expression level of E-cadherin in metastatic tumors decreased significantly compared with that in primary tumors. However, little change was observed in the mRNA level of beta-catenin in different tumor tissues. Only 4 samples (19.1%) displayed gene promoter methylation of E-cadherin in primary tumor and 10 samples (47.6%) showed methylated form of E-cadherin. The gene promoter methylation of E-cadherin was more common in metastatic tumor than in primary tumor of NPC (P = 0.024). Only 2 (4.76%) of the 42 samples showed mutations in exon 3 of beta-catenin at 41 (T41A, ACC-->GCC) and codon 47 (S47T, AGT-->ACT). The cytoplasmic and nuclear expression of beta-catenin in tumor was not found in any samples of NPC.

CONCLUSIONSThe results suggest that the downregulation of E-cadherin results from the gene promoter aberrant methylation of E-cadherin and that the methylation of E-cadherin plays an important role in invasion and metastasis of tumor cells in NPC. However, beta-catenin mutation is an infrequent event in NPC, and beta-catenin is not a critical factor influencing the invasion and metastasis of tumor cells in NPC.

Adult ; Aged ; Blotting, Western ; Cadherins ; analysis ; genetics ; Cytoskeletal Proteins ; analysis ; genetics ; DNA Methylation ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mutation ; Nasopharyngeal Neoplasms ; chemistry ; genetics ; pathology ; Neoplasm Metastasis ; Promoter Regions, Genetic ; Trans-Activators ; analysis ; genetics ; beta Catenin