17-allylamino-17-demethoxygeldanamycin (17-AAG) is a heat shock protein 90 ( HSP90) inhibitor. It is capable of target-inhibiting tumor-derived HSP90, leading to the depletion of on-cogenic client proteins which play key roles in several signal transduction pathways. Then cell growth and differetiation are inhibited. 17-AAG results in cytostasis and apoptosis. A lot of trials have indicated that 17-AAG is a selective target drug for cancer therapy. It can efficiently inhibit multiple signal transduction pathways involved in tumor cell proliferation and survival. Clinical phaseⅠand phaseⅡtrials have shown that 17-AAG has good pharmacokinetic-pharmacodynamic profile. Moreover, it can combine with radiation and the traditional chemotherapeutics and increase the therapeutic efficacy.

In order to explored the change of PML/PML-RARalpha protein during tetraarsenic tetrasulfide (As4S4) treatment, acute promyelocytic leukemia (APL) cells from a group of newly diagnosed APL patients were examined by indirect immunofluorescence staining with anit-PML monoclonal antibody. The results showed that all samples typically presented many microspeckle signals throughout the nucleus before treatment. The redistribution occurred as early as on the second day after As4S4 treatment, which revealed loss of microspeckles with the presentation of a few large speckles. Anti-PML staining also emerged in the perinuclear cytoplasm. At last, microspeckles and large speckles all disappeared. When the therapy was combining all-trans-retinoic acid (ATRA) with As4S4, similar results were obtained. However, APL cells from patients treated with ATRA alone performed totally different appearance, presenting microspeckles and large speckles at the same time, followed with entirely large speckles. The conclusion is that As4S4 makes redistribution of PML/PML-RARalpha protein in leukemic cells from APL patients during the treatment, which is quite different from that during the treatment of ATRA.
Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Arsenicals ; therapeutic use ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; In Situ Nick-End Labeling ; Leukemia, Promyelocytic, Acute ; drug therapy ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Oncogene Proteins, Fusion ; analysis ; Promyelocytic Leukemia Protein ; Transcription Factors ; analysis ; Tretinoin ; therapeutic use ; Tumor Suppressor Proteins

OBJECTIVETo investigate the clinical significance of the operation score system for endoscopic thyroidectomy.

METHODSAn operation score system based on 6 important procedure skills of endoscopic thyroidectomy was established. And a retrospective study of the first 300 consecutive patients underwent endoscopic thyroidectomy from July 2001 to December 2007 by a single surgeon was performed. The patients was divided into 10 consecutive groups chronologically, each comprising 30 cases.

RESULTSThe mean operation score of all the patients was 6.0 and the mean operation time was 98.1 min. There were significant differences in the mean operation score, every skill score and the mean operation time among the 10 groups. In the consecutive two groups comparison, significant differences in the operation scores were observed between group 2 and 3 (P < 0.05) and between group 5 and 6 (P < 0.05).

CONCLUSIONThe operation score system for endoscopy thyroidectomy is a useful method to judge the proficiency and the stability of the operation.

Adolescent ; Adult ; Endoscopy ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Outcome and Process Assessment (Health Care) ; Retrospective Studies ; Thyroidectomy ; Young Adult

OBJECTIVETo screen primary aldosteronism cases with ARR (aldosterone/plasma renin activity, ARR) from patients with hypertension, and to evaluate the diagnosis value of ARR in primary aldosteronism cases and analysis the clinical characters of primary aldosteronism cases.

METHODSNine hundred and two patients with hypertension were collected, the plasma aldosterone concentration to plasma renin activity ratio were detected by radio-immunity method, after that, ARR were calculated. Retrospective analysis was made of clinical data in 126 primary aldosteronism cases, which ARR were over 25.

RESULTSOne hundred and twenty-six cases (14%) were diagnosed as primary aldosteronism, and of them, 49 cases had hypokalemia. 25 patients received surgical operation and the rate of efficiency and cure of surgery treatment were 100% and 48%, respectively. The rate of efficiency and cure of drug treatment was 89% and 24% respectively.

CONCLUSIONSPrimary aldosteronism affects over 10% of patients with hypertension in China. Patients with hypertension and most patients with treatment-resistant hypertension should undergo screening for primary aldosteronism with ARR. A high ARR is a positive screening test result, a finding that warrants confirmatory testing.

Aldosterone ; blood ; Clinical Chemistry Tests ; Follow-Up Studies ; Humans ; Hyperaldosteronism ; diagnosis ; Hypertension ; blood ; Male ; Middle Aged ; Potassium ; blood ; Renin ; blood ; Renin-Angiotensin System

Objective: To investigate the characteristic and prognostic significance of leukemia stem cells associated antigens expressions including CD34, CD38, CD123, CD96 and TIM-3 in t (8;21) AML. Methods: Bone marrow samples of 47 t (8;21) AML patients were collected at diagnosis from October 2015 to April 2018 in Peking University Peoples' Hospital, then flow cytometry method was performed to detect the expression frequencies of CD34, CD38, CD123, CD96 and TIM-3 to analyze the relationship between leukemia stem cells associated antigens expressions and relapse. Results: Of 47 t (8;21) AML patients tested, the median percentages of CD34(+)CD38(-), CD34(+) CD38(-)CD123(+), CD34(+)CD38(-) CD96(+) and CD34(+) CD38(-) TIM-3(+) cells among nucleated cells were 2.37%, 0.24%, 0.27% and 0.06%, respectively. All the frequencies of CD34(+)CD38(-), CD34(+)CD38(-)CD123(+), CD34(+)CD38(-)CD96(+) and CD34(+) CD38(-)TIM-3(+) cells had no impact on the achievement of CR after the first course of induction. All higher frequencies of CD34(+)CD38(-), CD34(+)CD38(-)CD123(+), CD34(+)CD38(-)CD96(+) cells were related to higher 2-year CIR rate. Whereas, the frequency of CD34(+) CD38(-) TIM-3(+) cells had no impact on CIR rate. Both high frequency of CD34(+) CD38(-) cells and the high level of minimal residual diseases (patients with <3-log reduction in the RUNX1-RUNX1T1 transcript level after the second consolidation therapy) were independent poor prognostic factors of CIR[P=0.025, HR=6.9 (95%CI 1.3-37.4) ; P=0.031, HR=11.1 (95%CI 1.2-99.2) ]. Conclusion: Different leukemia stem cells associated antigens had distinct prognostic significance in t (8;21) AML. High frequencies of CD34(+) CD38(-), CD34(+) CD38(-) CD123(+) and CD34(+)CD38(-)CD96(+) cells at diagnosis predicted relapse in patients with t (8;21) AML.
ADP-ribosyl Cyclase 1 ; Antigens, CD ; Flow Cytometry ; Humans ; Interleukin-3 Receptor alpha Subunit ; Leukemia, Myeloid, Acute ; Neoplastic Stem Cells ; Prognosis ; Stem Cells

OBJECTIVETo explore the specificity of anti-phosphotyrosine monoclonal antibody PY20 in bcr-abl+ cells and its possible clinical applications.

METHODSBcr-abl cell lines( K562, MEG-01) and bcr-abl- cells lines( Jurkat, MCF-7 )were stained with PY20. Phosphotyrosine protein of K562 and MEG-01 cells was detected by flow cytometry before and after treatment with imatinib. Phosphotyrosine protein in bone marrow cells from 49 patients with chronic myeloid leukemia (CML), Ph+ acute lymphoblastic leukemia(Ph(+) -ALL), Ph- ALL, acute myeloid leukemia (AML-M1, M2, M3, M5, FAB classification), chronic lymphocytic leukemia (CML) and 3 normal donor. Positive cells over 5% of total cells was considered positive cases for phosphotyrosine protein. The level of tyrosine phosphorylation was determined by median fluorescence intensity (MFI).

RESULTSBcr-abl cell lines and marrow cells from 10 CML patients and 8 ALL patients were all PY20-positive, while bcr-abl- cell lines and marrow cells from 18 leukemia patients and 3 normal donor were all PY20-negative. MFI decreased remarkably after blocked by imatinib in K562 cells and MEG-01 cells. The positive cell percent of marrow cells from 10 newly diagnosed CML patients and 9 imatinib-sensitive CML patients was (54.20 +/- 19.82)% and (14.84 +/- 6.17)% (P < 0.05), while that of 2 cases of imatinib-resistant was 64.3% and 57.2%. There was significant difference of MFI between imatinib-resistant patients and imatinib-sensitive patients (99.42 +/- 4.87 vs 46.41 +/- 4.67, P < 0.01).

CONCLUSIONPY20 monoclonal antibody is highly specific for bcr-abl+ cells. It might be useful in rapid detection of bcr-abl+ cells and sensitivity to imatinib of CML patients.

Antibodies, Monoclonal ; analysis ; Bone Marrow Cells ; metabolism ; Flow Cytometry ; Fusion Proteins, bcr-abl ; analysis ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Phosphotyrosine ; analysis ; immunology

BACKGROUNDReal-time quantitative RT-PCR (RQ-PCR) assay has become a vital tool to monitor residual disease of leukemia. However, the complexity and standardization of RQ-PCR should never be overlooked and the results should be interpreted cautiously in clinical conditions. We aimed to assess the methodology of RQ-PCR and its clinical applications in monitoring molecular kinetics of 36 newly diagnosed cases of acute promyelocytic leukemia patients with t (15; 17) from October 2004 to December 2005.

METHODSAll the TaqMan probe-based RQ-PCR reactions and analysis were performed on an ABI-PRISM 7,500 platform. The quantitation of PML-RARalpha transcripts was represented by the normalized quotient, that is, PML-RARalpha transcript copies divided by ABL transcript copies. According to induction therapy, the patients were classed into two groups: group 1 (n = 23), three-drug combination including arsenics, all-trans retinoic acid and mitoxantrone; and group 2 (n = 13), two-drug combination from all-trans retinoic acid, arsenics and mitoxantrone.

RESULTSThe sensitivity of RQ-PCR was 1 per 10(5) cells and 5 copies of the PML-RARalpha transcript could be reproducibly detected. No false positive results occurred in 40 non-acute promyelocytic leukemia samples. Optimal amplification efficiency could be attained, which was determined by the slope of the standard curves (slope: -3.2 - -3.7). The inter-assay and intra-assay variation coefficients of the method were 1.01% and 0.56% respectively. Although the time to attain hematological complete remission was similar in both groups, the time to achieve molecular remission of group 1 was significantly shorter than that of group 2 (61 days vs 75 days, P = 0.034). The rate of molecular remission within 70 days was higher in group 1 than in group 2 (75.00% vs 38.46%, P = 0.036). Compared with pretreatment, median reduction of the PML-RARalpha transcript before first consolidation therapy differed significantly between group 1 and group 2 (log scale, 3.15 vs 2.31, P = 0.024). Interestingly, we found that PML-RARalpha transcript levels temporarily increased in bone marrow (7 patients) and peripheral blood (22 patients) samples of patients during induction therapy in both groups.

CONCLUSIONSThe RQ-PCR assay is reliable for the detection of PML-RARalpha transcripts. Arsenics, all-trans retinoic acid and mitoxantrone triad induction treatment of acute promyelocytic leukemia is superior to two-drug combination induction therapy in terms of the molecular response.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Child ; Female ; Humans ; Leukemia, Promyelocytic, Acute ; blood ; drug therapy ; genetics ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

The aim of study was to investigate the importance of chromosome aberration in differential diagnosis of eosinophilia and the chromosomal aberrations involved in patients with clonal eosinophilia. 65 cases of eosinophilia were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture, and G-banding technique was used for karyotyping. The results showed that out of 65 cases, chromosome 16 inversion was detected in 9 patients suspected as M(4Eo), and among the other 56 cases, 5 were detected with chromosomal aberrations (8.9%). Combining clinical, hematological and cytogenetical data, the 5 patients were diagnosed as acute myeloid leukemia with eosinophilia, chronic eosinophilic leukemia, 8p11 myeloproliferative syndrome, chronic myeloid leukemia in acute phase and acute myeloid leukemia-M(4Eo) respectively. The detected chromosomal aberrations were +14, t (5; 12) (q31; p13), t (8; 9) (p11; q32), t (9; 22) (q34; q11) and inv (16) (p13 q22). In conclusion, cytogenetical detection is very important in differential diagnosis of clonal eosinophilic disorders and chronic eosinophilic leukemia, which is suggested to be done routinely in clinic.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chromosome Aberrations ; Chromosomes, Human, Pair 16 ; genetics ; Cytogenetic Analysis ; Diagnosis, Differential ; Eosinophilia ; diagnosis ; genetics ; pathology ; Female ; Humans ; Hypereosinophilic Syndrome ; diagnosis ; genetics ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the relationship between serum 25-hydroxyvitamin D [25(OH)D] levels at birth and respiratory distress syndrome (RDS) in preterm infants.

METHODSThis retrospective study recruited preterm infants with gestational age of below 34 weeks who were born between January 2014 and December 2016. These preterm infants were divided into two groups: RDS (n=72) and control (n=40). Clinical data of the two groups were collected, including gestational age, birth weight, gender, delivery mode, Apgar scores at 1 minute and 5 minutes, incidence of maternal gestational diabetes mellitus, and use of prenatal steroid hormone. Peripheral blood samples were collected and 25(OH)D levels were measured by chemiluminescence immunoassay. The association between serum 25(OH)D levels at birth and RDS was analyzed by multivariate logistic regression.

RESULTSApgar scores at 1 minute and 5 minutes and serum 25(OH)D levels in the RDS group were significantly lower than those in the control group (P<0.05), while the rates of neonatal asphyxia and vitamin D deficiency were significantly higher than those in the control group (P<0.05). Multivariate logistic regression analysis showed that neonatal asphyxia (OR=2.633, 95%CI: 1.139-6.085) and vitamin D deficiency (OR=4.064, 95%CI: 1.625-10.165) were risk factors for RDS in preterm infants.

CONCLUSIONSVitamin D deficiency might be associated with increased risk of RDS in preterm infants. Reasonable vitamin D supplementation during pregnancy might reduce the incidence of RDS in preterm infants.

Dietary Supplements ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Respiratory Distress Syndrome, Newborn ; blood ; etiology ; Vitamin D ; analogs & derivatives ; blood ; Vitamin D Deficiency ; complications

OBJECTIVETo assess the association between serum 25-hydroxyvitamin D [25(OH)D] levels at birth and bronchopulmonary dysplasia (BPD) in preterm infants.

METHODSThis study recruited preterm infants with gestational age of below 34 weeks who were born between January 2014 and December 2016. These preterm infants were classified into two groups: BPD and control. The association between serum 25(OH)D levels at birth and BPD was analyzed.

RESULTSSerum 25(OH)D levels in the BPD group was significantly lower than those in the control group [(37±17 nmol/L vs 47±20 nmol/L; P<0.05), and the rate of vitamin D deficiency was significantly higher than those in the control group (90.2% vs 74.0%; P<0.05). The level of serum 25(OH)D was negatively correlated with the incidence of BPD (r=-0.201, P=0.001).

CONCLUSIONSVitamin D deficiency at birth may be associated with BPD in preterm infants, but need to be further studied by multivariate analysis.

Bronchopulmonary Dysplasia ; blood ; etiology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; blood ; Male ; Vitamin D ; analogs & derivatives ; blood ; Vitamin D Deficiency ; complications