This study was purposed to investigate the methylation status of id4 gene promoter in patients with chronic myeloid leukemia (CML) and explore the relationship between methylation of the id4 gene and progress of CML. The methylation status of id4 gene in 48 chronic myeloid leukemia patients and 10 healthy individuals was detected by using methylation-specific polymerase chain reaction (MS-PCR). The results showed that id4 gene was unmethylated in bone marrow samples from both healthy individuals and CML patients in chronic phase (CP). The rate of id4 gene methylation in both CML patients in accelerated phase (AP) and blast crisis (BC) was 66%, and was higher than those of CML patients in CP phase. There was significant difference between them (p < 0.05). In one CML patient who received a serial observations, the status of id4 was unmethylated in CP, but it was methylated in AP and BC phase. It is concluded that the id4 gene in CML patients is unmethylated in CP, while it is methylated in AP or BC. The detection of id4 gene methylation status may be useful for monitoring disease advance in CML and may be used as a marker of disease progression in CML.
Adolescent ; Adult ; Aged ; Child ; DNA Methylation ; DNA Primers ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Promoter Regions, Genetic ; Young Adult

Objective To explore the application value of spectral CT quantitative analysis in differentiating adenocarcinoma or squamous carcinoma from inflammatory myofibroblastic tumor (IMT). Methods A total of 115 patients with 62 adenocarcinomas, 33 squamous carcinomas and 20 IMTs underwent spectral CT scans to obtain spectral images at arterial phase (AP) and venous phase (VP). The imaging data were analyzed retrospectively. The iodine concentration of adenocarcinoma, squamous carcinomas and IMT were measured. The normalized iodine concentration in AP (NICAP), normalized iodine concentration in VP (NICVP) and normalized iodine concentration difference between AP and VP (ICD) were calculated. The above quantitative parameters among three groups were analyzed with analysis of variance and ROC curve. Results NICAP (0.15 ± 0.04), NICVP (0.37 ± 0.08) and ICD(0.23 ± 0.06)of the adenocarcinoma were lower than those of IMT (0.21 ± 0.05,0.50 ± 0.06,0.28 ± 0.08). There were significant differences in NICAP, NICVP and ICD between adenocarcinoma and IMT (P<0.05). NICAP (0.13 ± 0.03), NICVP (0.35±0.06) and ICD (0.22±0.05) of the squamous carcinoma were lower than those of IMT (0.21± 0.05,0.50±0.06,0.28±0.08). The differences in NICAP, NICVP and ICD were significant between squamous carcinoma and IMT (P<0.05). There were no significant differences in NICAP, NICVP and ICD between adenocarcinoma and squamous carcinoma (P>0.05). The best spectral quantitative parameter for differentiating the adenocarcinoma from IMT was NICVP, which yielded a sensitivity of 92.3% and a specificity of 86.7%with the threshold of 0.425. NICVP was also the best spectral quantitative parameter for differentiating squamous carcinomas from IMT. With the threshold of 0.44, a sensitivity of 84.6% and a specificity of 92.3% were found. Conclusion Spectral CT imaging with the quantitative iodine concentration analysis may help to increase the accuracy of differentiating adenocarcinoma and squamous carcinoma from IMT.

Objective To explore the application value of spectral CT quantitative analysis in differentiating adenocarcinoma or squamous carcinoma from inflammatory myofibroblastic tumor (IMT). Methods A total of 115 patients with 62 adenocarcinomas, 33 squamous carcinomas and 20 IMTs underwent spectral CT scans to obtain spectral images at arterial phase (AP) and venous phase (VP). The imaging data were analyzed retrospectively. The iodine concentration of adenocarcinoma, squamous carcinomas and IMT were measured. The normalized iodine concentration in AP (NICAP), normalized iodine concentration in VP (NICVP) and normalized iodine concentration difference between AP and VP (ICD) were calculated. The above quantitative parameters among three groups were analyzed with analysis of variance and ROC curve. Results NICAP (0.15 ± 0.04), NICVP (0.37 ± 0.08) and ICD(0.23 ± 0.06)of the adenocarcinoma were lower than those of IMT (0.21 ± 0.05,0.50 ± 0.06,0.28 ± 0.08). There were significant differences in NICAP, NICVP and ICD between adenocarcinoma and IMT (P<0.05). NICAP (0.13 ± 0.03), NICVP (0.35±0.06) and ICD (0.22±0.05) of the squamous carcinoma were lower than those of IMT (0.21± 0.05,0.50±0.06,0.28±0.08). The differences in NICAP, NICVP and ICD were significant between squamous carcinoma and IMT (P<0.05). There were no significant differences in NICAP, NICVP and ICD between adenocarcinoma and squamous carcinoma (P>0.05). The best spectral quantitative parameter for differentiating the adenocarcinoma from IMT was NICVP, which yielded a sensitivity of 92.3% and a specificity of 86.7%with the threshold of 0.425. NICVP was also the best spectral quantitative parameter for differentiating squamous carcinomas from IMT. With the threshold of 0.44, a sensitivity of 84.6% and a specificity of 92.3% were found. Conclusion Spectral CT imaging with the quantitative iodine concentration analysis may help to increase the accuracy of differentiating adenocarcinoma and squamous carcinoma from IMT.

It is hard to discriminate myelodysplastic syndrome(MDS) from many benign hematological diseases. To identify the methylation status of zo-1 gene in MDS, the methylation specific PCR (MS-PCR) and reverse transcription-PCR (RT-PCR) were applied to detect the MDS cell line MUTZ-1, bone marrow of a healthy donor and an aplastic anemia patient. MS-PCR was also employed to detect the bone marrow of 72 patients with benign hematological diseases, 35 MDS-RA patients, and 20 MDS-like patients. The results showed that MDS cell line MUTZ-1 displayed complete methylation of zo-1 promoter without mRNA expression. Inversely, a patient with benign hematological disease and a donor with normal bone marrow showed complete unmethylation of this gene with unaffected mRNA expression. No zo-1 promoter methylation was detected in patients with benign hematological diseases, while aberrant hypermethylation of zo-1 gene promoter were found in 48.6% (18/37) of MDS-RA patients. The positive rate of zo-1 methylation in MDS-RA patients was higher than that in patients with benign hematological diseases (p < 0.05). Seven suspected MDS patients manifested hypermethylation status of zo-1 gene (7/20), 2 were followed up for 1 year and transformed into MDS. It is concluded that relatively high hypermethylation rate of zo-1 promoter is observed in MDS-RA, and no methylation in patients with benign hematological diseases. Therefore, zo-1 gene hypermethylation may be served as a useful epigenetic marker in the differential diagnosis for MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA Methylation ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; genetics ; Young Adult ; Zonula Occludens-1 Protein ; genetics ; metabolism

Multiple myeloma (MM) is an incurable heterogeneous disease derived from malignant clonal expansion of plasma cells. The evaluation of prognosis, detection of minimal residual disease (MRD) and treatment of MM are unclear for decades. Recently, Velcade and autotransplantation have been broadly applied to MM patients and achieved better outcomes, but there is yet no effective and universal marker for MRD detection in MM. Both genetic and epigene-tic aberrations play important roles in the pathogenesis and development of cancer. Our preliminary data showed that aberrant promoter methylation of zo-1 and id4 genes was correlated with their gene silencing in several types of hematological malignancies. Therefore, this study was aimed to identify the promoter methylation status of zo-1 and id4 genes in MM and their relationship with the prognosis, MRD and treatment of MM. The methylation status of zo-1 and id4 genes of MM cell lines U266, H929 and IM9 was tested by using MS-PCR; the methylation status of zo-1, id4 gene promoters in bone marrow samples of 20 MM patients and 6 healthy persons was detected by MS-PCR. The results showed that the zo-1, id4 gene in MM cell lines all were methylation positive (complete or partial methylation), the zo-1, id4 gene in samples of 5 healthy persons all were completely unmethylated. The methylation positive rate of zo-1 and id4 genes were 50% and 85% respectively, which were significantly higher than that in normal bone marrow (0%). The coverage rate of zo-1 and id4 gene methylation was 95%. There were no significant differences in the methylation status of both genes among the patients with different heavy chains, different light chains and symptoms. It is concluded that the change of zo-1, id4 genes methylation status occurs in MM patients and has specificity, which may be a new gene marker of MM, methylation analysis of zo-1 and id4 genes may be important for MRD monitoring in patients with MM.
Adult ; Aged ; Bone Marrow ; metabolism ; Cell Line, Tumor ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; Neoplasm, Residual ; genetics ; Phosphoproteins ; genetics ; metabolism ; Zonula Occludens-1 Protein

BACKGROUNDThe diagnosis of myelodysplastic syndrome (MDS), especially hypoplastic MDS, and MDS with low blast counts or normal karyotype may be problematic. This study characterized ID4 gene methylation in patients with MDS and aplastic anemia (AA).

METHODSThe methylation status of ID4 was analyzed by bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time methylation-specific PCR (MethyLight PCR) in 100 patients with MDS and 31 patients with AA.

RESULTSThe MDS group had a higher ID4 gene methylation positivity rate (22.22%) and higher methylation levels (0.21 [0-3.79]) than the AA group (P < 0.05). Furthermore, there were significant differences between the hypoplastic MDS and AA groups, the MDS with low blast count and the AA groups, and the MDS with normal karyotype and the AA groups. The combination of genetic and epigenetic markers was used in much more patients with MDS (62.5% [35/56]) than the use of genetic markers only (51.79% [29/56]).

CONCLUSIONSThese results showed that the detection of ID4 methylation positivity rates and levels could be a useful biomarker for MDS diagnosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Aplastic ; genetics ; Child ; CpG Islands ; genetics ; DNA Methylation ; genetics ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Young Adult

OBJECTIVEIn order to develop plans for effective intervention measures, prevalence and health-seeking behavior related to reproductive tract infection among floating married women of childbearing age in Fengtai district in Beijing were studied.

METHODSCross-sectional study was carried out. Two thousand and sixty-nine eligible women were randomly selected from strata based on their home provinces. From June to July 2001, the subjects were given face-to-face interview at the Fengtai family planning clinic in Beijing using standard questionnaire followed by gynecologic examination and laboratory tests.

RESULTSThirty point three percent of the subjects were found to have reproductive tract infections (RTI) by laboratory tests. Prevalence rates of bacterial vaginosis, candida and trichomonas vaginitis were 22.2%, 4.9% and 2.1% respectively. Prevalence rates of chlamydia, gonorrhea, condyloma acuminatum and syphilis were 2.2%, 1.6%, 0.5% and 0.2% respectively. Of these infected women, only 43.1% (270/626) were symptomatic, and 61.5% (166/270) of these women with symptoms had sought treatment.

CONCLUSIONCompared to other results in the literature, we found a relatively high prevalence of RTI in our study population. Only a small proportion of these infected women were symptomatic but only few of them sought treatment. We suggested that the provision of more family planning service and promotion of RTI knowledge to the floating women of childbearing age.

Adolescent ; Adult ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Infection ; epidemiology ; Middle Aged ; Prevalence ; Surveys and Questionnaires ; Travel ; Trichomonas Vaginitis ; epidemiology ; Urban Health ; Vaginitis ; epidemiology ; microbiology ; Vaginosis, Bacterial ; epidemiology ; Women's Health Services

The objective was to observe the influence of previously activated psoralens on the proliferation of K562 cells, and to provide laboratory data for its clinical usage. K562 cells were treated separately with previously and late activated psoralens, then their trypan blue exclusion inhibited rates (TBIR), cell proliferation inhibited rates (CPIR) and colony forming inhibited rates (CFIR) after culture were compared. The results showed that previously activated psoralens displayed an inhibiting effect on the proliferation of K562 cells with a dose-effect relationship. There was no obvious difference between previously and late activated psoralens on TBIR, CPIR and CFIR. In order to exert the inhibitive effect of previously activated psoralens, the time of ultraviolet ray exposure should be 10 minutes at least, and longer than 12 hours for inhibiting K562. The inhibitive effect of previously activated psoratens decreased as the time interval from activation to its use was prolonged. The inhibiting effect of previously activated psoralens was strongest within 6 hours after activation. In conclusion, both previously and late activated psoralens show inhibiting effects on the proliferation of K562, which may be able to use an antineoplastic drug in clinic.
Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Furocoumarins ; pharmacology ; Humans ; K562 Cells ; Time Factors

Objective To summarize the epidemiological and clinical features of infectious diseases of the central nervous system(CNS)by a single-center analysis.Methods A retrospective analysis was conducted on the data of 1247 cases of CNS infectious diseases diagnosed and treated in the First Medical Center of PLA General Hospital from 2001 to 2020.Results The data for this group of CNS infectious diseases by disease type in descending order of number of cases were viruses 743(59.6％),Mycobacterium tuberculosis 249(20.0％),other bacteria 150(12.0％),fungi 68(5.5％),parasites 18(1.4％),Treponema pallidum 18(1.4％)and rickettsia 1(0.1％).The number of cases increased by 177 cases(33.1％)in the latter 10 years compared to the previous 10 years(P<0.05).No significant difference in seasonal distribution pattern of data between disease types(P>0.05).Male to female ratio is 1.87︰1,mostly under 60 years of age.Viruses are more likely to infect students,most often at university/college level and above,farmers are overrepresented among bacteria and Mycobacterium tuberculosis,and more infections of Treponema pallidum in workers.CNS infectious diseases are characterized by fever,headache and signs of meningeal irritation,with the adductor nerve being the more commonly involved cranial nerve.Matagenomic next-generation sequencing improves clinical diagnostic capabilities.The median hospital days for CNS infectious diseases are 18.00(11.00,27.00)and median hospital costs are ￥29,500(￥16,000,￥59,200).The mortality rate from CNS infectious diseases is 1.6％.Conclusions The incidence of CNS infectious diseases is increasing last ten years,with complex clinical presentation,severe symptoms and poor prognosis.Early and accurate diagnosis and standardized clinical treatment can significantly reduce the morbidity and mortality rate and ease the burden of disease.

OBJECTIVETo evaluate the prevalence of Fms-Like tyrosine kinase 3 (FLT3) mutations including internal tandem duplication (ITD) of juxtamembrane region and point mutation in the second tyrosine kinase domain (TKD) in acute promyelocytic leukemia (APL) and its clinical significance.

METHODSBone marrow mononuclear cells from 160 newly diagnosed APL patients were analyzed. Polymerase chain reaction (PCR) was used to detect FLT3-ITD mutations, FLT3-ITD positive samples were further analyzed for the ITD allelic ratio (ITD-AR, mutant-wild type ratio). The FLT3-TKD mutation was analyzed by PCR amplification of exon 20 followed by EcoR V digestion and sequencing.

RESULTSOut of 160 patients, 30 (18.75%) patients were FLT3-ITD positive, 17 (10.62%) were FLT3-TKD positive, 2 had both of mutations. The initial WBC count and the ratio of short type PML-RAR alpha isoforms in FLT3-ITD positive and FLT3-TKD positive patients were all higher than that in patients with wild-type FLT3 (FLT3-wt) (P < 0.05). For FLT3-ITD positive patients, the incidences of retinoic acid syndrome (RAS) and disseminated intravascular coagulation (DIC) were 41.7% and 65.4%, respectively, being higher than that of FLT3-wt patients, while their complete remission (CR) rate was lower (69.2% vs 90.3%, P < 0.05). For FLT3-TKD positive patients, the incidence of RAS, DIC and CR rate were not significantly different from that of FLT3-wt patients (P > 0.05). FLT3-ITD positive patients had a shorter overall survival (OS) (P < 0.05), but not disease-free survival (DFS) (P > 0.05) as compared with FLT3-wt patients. There was no significant difference in either OS or DFS between FLT3-TKD positive and FLT3-wt patients. The ITD-AR of 30 FLT3-ITD positive patients varied from 0.11 to 6.55 with a median of 1.0. The initial WBC count, incidence of RAS and DIC, CR rate were not significantly different between the patients with ITD-AR greater than 1.0 and lower than 1.0 (P > 0.05).

CONCLUSIONSFLT3 mutations (FLT3-ITD or FLT3-TKD) are frequently identified in patients with newly diagnosed APL, both mutations are associated with higher initial WBC and short type PML-RAR alpha isoforms. FLT3-ITD mutation is more frequent than FLT3-TKD mutation, and predicts a poorer prognosis, whereas FLT3-TKD mutation does not show the same unfavorable prognostic effect on APL patients.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Point Mutation ; Prognosis ; Tandem Repeat Sequences ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics