Discovery, biosynthesis, organic synthesis, and bioactivities of meroterpenoids from Rhododendron species.

Meroterpenoids discovered in Rhododendrons species possess unique chemical structures and biological activities and are expected to become new drug targets for Alzheimer's disease, metabolic disorders, and chronic kidney disease, and these compounds have attracted increasing attention in recent years. In this study, Rhododendron meroterpenoids and their structures, classifications, racemate distribution, biosynthetic pathways, chemical synthesis, and bioactivities are reviewed prior to 2023.

The association between multimorbidity and health-related quality of life: a cross-sectional survey among community middle-aged and elderly residents in southern China.

BACKGROUND: Multimorbidity is common among the middle-aged and elderly residents. And it is associated to the reduction of health-related quality of life (HRQoL), including physical and psychological dimensions. However, there are few studies that have paid attention to the HRQoL of residents with multimorbidity in China. Therefore, this study aims to investigate the relationships between different multimorbidity patterns and HRQoL among middle-aged and elderly adults in China. METHODS: Based on a cross-sectional survey, the information regarding 18,137 adults, who were at least 45 years of age, was collected through interviews. Self-perceived HRQoL was assessed with the EQ-5D-3 L instrument, and the EQ-5D-3 L index score was calculated using the Chinese EQ-5D-3 L value set. The Tobit regression was used to explore the impacts of multimorbidity groups on HRQoL. RESULTS: Of 18,137 respondents, more than a fifth (3773,20.8%) of people had multimorbidity. Mean (SD) of EQ-5D index and VAS values were 0.95(0.14) and 76.02(13.66), respectively. Significant correlations were found between a lower HRQoL and increasing numbers of chronic conditions (P < 0.001). Most of chronic diseases co-occurred frequently, and the association between hypertension and diabetes mellitus was the strongest (adjusted OR = 3.82). The most prevalent disease is hypertension (5052,27.9%), and the most prevalent chronic diseases pair is hypertension and diabetes mellitus (841,4.6%). Among those chronic diseases with high prevalence, the effects on HRQoL ranged from chronic pain to hypertension (adjust b = - 0.036 to - 0.008). In the common multimorbidity patterns, co-occurrence of chronic pain and bone disease (adjust b = - 0.039) had the greatest impact on HRQoL. CONCLUSIONS: The HRQoL of middle-aged and elderly adults declines by multimorbidity. More attention should be paid to the HRQoL of residents with multimorbidity in China. The effect of different multimorbidity patterns on HRQoL is not simply added by two diseases, but changes by the different combination. Identifying different multimorbidity patterns of residents can provide more targeted measures to improve the HRQoL.

Association between obesity and sickness in the past two weeks among middle-aged and elderly women: A cross-sectional study in Southern China.

OBJECTIVES: Sickness situation in the past two weeks, an indicator of health service needs, is an increasing major health concern. However, data on the relationship between obesity and two-week morbidity in the female population, particularly in middle-aged and elderly women, is sparse. The present study aimed to examine the association between obesity and two-week morbidity among middle-aged and elderly women in Southern China, and to explore the independent contributions of socio-demographic variables, health-related factors, and obesity to two-week morbidity. METHODS: In total, 2364 middle-aged and elderly women were included in this cross-sectional, community-based survey. Obesity was assessed using body mass index (BMI). The outcome variable was sickness situation over the past two weeks (two-week morbidity). Clustered logistic regression was applied to analyze the independent contribution of obesity to two-week morbidity. RESULTS: Approximately 14.6% of participants experienced sickness in the past two weeks. Obesity (odds ratio [OR] = 1.47, 95% confidence interval [CI] = 1.02-2.12) was significantly associated with two-week morbidity and its independent contribution accounted for 3.7%, lower than that of socio-demographic variables (73.7%) and health-related factors(22.6%). CONCLUSIONS: Some degree of correlation was observed between obesity and two-week morbidity among middle-aged and elderly women in Southern China, which can be used as a reference for health-related decision-making.

[Mechanism of Platycarya strobilacea Sieb. et Zucc extract-induced methuosis in human nasopharyngeal carcinoma CNE1 and CNE2 cells].

OBJECTIVE: To study the effect of Platycarya strobilacea Sieb. et Zucc (PSZ) extract on methuosis of human nasopharyngeal carcinoma CNE1 and CNE2 cells and explore the underlying mechanism. METHODS: CNE1 and CNE2 cells were treated with 1 mg/mL PSZ extract and the expressions of Rac1 mRNA and Rac1 protein were detected using RT-qPCR and Western blotting, respectively. Results CNE1 and CNE2 cells showed obvious morphological changes typical of methuosis following treatment with PSZ extract characterized by cell merging, accumulation of large cytoplasmic vacuoles, and membrane rupture without obvious changes in the nuclei. PSZ treatment resulted in up-regulated Rac1 mRNA and Rac1 protein expressions in the cells. Application of EHT 1864 obviously blocked the effect of PSZ extract in inducing methuosis in CNE1 and CNE2 cells. CONCLUSION: PSZ extract can induce methuosis in CNE1 and CNE2 cells by inducing the overexpression of Rac1.

EphB4-VAV1 signaling pathway is associated with imatinib resistance in chronic myeloid leukemia cells.

Imatinib (IM) resistant Chronic Myeloid Leukemia (CML) is an important issue to be addressed while treating CML patients. The present study analyzes the role of EphB4-VAV1 signaling in IM-resistant CML. EphB4 and VAV1 were overexpressed in IM-resistant CML patients and K562-R cell line (K562-R). Then, we established stable under-expressing EphB4 cell line K562-R-EphB4-sh. VAV1 was down-regulated in K562-R-EphB4-sh cells. K562-R-EphB4-sh cells gained re-sensitivity to IM and K562-R cells showed mild IM resistance. However, EphB4 was no changed when the VAV1 was down-regulated. EphB4 and VAV1 were overexpressed in IM-resistant CML, VAV1might be the downstream moleculars of EphB4. These results suggest a potential role of EphB4-VAV1 signaling as therapeutic target of IM-resistant CML.

[Clinical Efficacy of Sorafenib Combined with Low Dose Cytarabine for Treating Patients with FLT3+ Relapsed and Refractory Acute Myeloid Leukemia].

OBJECTIVE: To study the efficacy and safety of sorafenib combined with low dose cytarabine for treating patients with FLT3(+) relapsed and refractory acute myeloid leukemia (FLT3(+) RR-AML). METHODS: Seven patients with FLT3(+) RR-AML were treated with sorafenib and low dose cytarabine. The curative rate and adverse effects were observed in these patients. RESULTS: Out of 7 RR-AML patients after treatment, 5 patients achieved complete remission (CR), 2 patients achieved partial remission (PR), and the overall response rate (ORR) after one course of therapy was 100%. No severe bleeding, nausea, vomiting and other side effects were found in these patients. CONCLUSION: Sorafenib combined with low dose cytarabine can effectively induce the remission of FLT3(+) RR-AML patients, and is worth for further clinical trails to verify its safty and efficiency.

CDKN2 Gene Deletion as Poor Prognosis Predictor Involved in the Progression of Adult B-Lineage Acute Lymphoblastic Leukemia Patients.

Deletion of cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) is well known in many hematologic malignancies, but only few reports have investigated this deletion effect on clinical prognosis. This study performed analysis of the CDKN2 deletion in 215 adult B- lineage acute lymphoblastic leukemia (B-ALL) patients, and related cytogenetic prognostic factors (BCR/ABL; E2A/PBXl; TEL/AML1; Mixed Lineage Leukemia (MLL) rearrangement; MYC, Immunoglobulin heavy locus (IGH) translocation). The prevalence of CDKN2 deletions in all study populations was 28.4%. There is no difference between patients with CDKN2 deletion and wild-type patients in sex, age, white blood cells (WBC) count, BM blast percentage, extra infiltration and induction complete remission (CR) rate. Analysis in relapse patients revealed that the distribution of CDKN2 deletion is higher in relapse patients (44.6%) than all patients (28.4%, P=0.006). Deletion of CDKN2 was significantly associated with poor outcomes including decreased overall survival (OS) (P<0.001), lower disease free-survival (DFS) (P<0.001), and increased cumulative incidence of relapse (P=0.002); Also, CDKN2 deletion was strongly associated with IGH translocation (P=0.021); and had an adverse effect on patients with BCR-ABL fusion gene or with MLL rearrangement. Patients with CDKN2 gene deletion benefited from allogenic hematopoietic stem cell transplantation (Allo-HSCT). Deletion of CDKN2 gene was commonly observed through leukemia progression and was poor prognostic marker in long-term outcomes.

[Control study on colonoscopy skills acquiring from endoscopic simulation system transfering to patients].

OBJECTIVE: To explore the feasibility of transferring the skills from the AccuTouch flexible endoscopy simulator colonoscopy training to clinical practices. METHODS: The novice colonoscopies were divided into 2 groups.Group A (control group) including 4 trainees for traditional training, Group B (experimental group) including 4 trainees for simulator training. After training, we compared the number of cases for achieving independent competence, assisted competence and incompetence in the first ten patients. RESULTS: No significant differences existed between two groups in terms of age and gender (both P > 0.05). Significant differences existed in educational background and the controlled group was better than the experimental group (Z = -2.005, P = 0.04). The cases of independent completion, assisted competence and incompetence of the control and experimental groups were 2, 4, 9 and 21, 29, 15 respectively. Rank tests show that the simulator training was better than the traditional counterpart (average rank: 56.14 vs 24.86, Z = -6.393, P = 0.00). CONCLUSIONS: The skills acquired from AccuTouch Endoscopy Simulator may be well transferred into the clinical colonoscopy environment. It clearly supports the scheme of integrating simulator training into colonoscopic education curricula.

[Application of multiprobe fluorescence in situ hybridization panel in the diagnosis of acute myeloid leukemia].

OBJECTIVE: To assess the value of multiprobe fluorescence in situ hybridization (FISH) panel in the diagnosis of acute myeloid leukemia (AML). METHODS: The multiprobe AML/MDS panel comprising 8 different FISH probes for AML1/ETO transfusion gene, PML-RARα transfusion gene, CBFß/MYH11 transfusion gene, MLL breakapart, P53 deletion, Del(5q), -7/Del(7q), and Del(20q) was tested in 40 cases of AML, and the results were compared with those by conventional cytogenetic G-banding (CCG) test. RESULTS: With multiprobe FISH panel, 22 of the 40 AML cases were found to carry 7 types of cytogenetic abnormalities, namely AML1/ETO transfusion gene, PML-RARα transfusion gene, MLL breakapart, P53 deletion, Del(5q), -7/Del(7q) and trisomy 8. The positive ratio of the multiprobe FISH was 57.5%. CCG only identified 8 cases with the corresponding cytogenetic abnormalities and 3 cases with other cytogenetic abnormalities, and the positive ratio was only 27.50%. CONCLUSION: Mutiprobe FISH panel is more rapid, accurate and effective than CCG in the diagnosis of AML.

[Multiprobe fluorescence in situ hybridization panel in detection of the common cytogenetic abnormalities of acute myeloid leukemia].

AIM: To evaluate the value of multiprobe Fluorescence in situ hybridization (FISH) panel in detection of the common cytogenetic abnormalities in acute myeloidleukemia( AML). And to investigate its association with clinical diagnosis, chemotherapy and prognosis. METHODS: Using the multiprobe AML/MDS panel designed to detect upto eight different FISH probes, which was for AML1/ETO transfusion gene, PML-RARα transfusion gene, CBFß/MYH11 transfusion gene, MLL breakapart, P53 deletion,Del(5q), Del(7q), Del(20q), 40 cases of AML were investigated. The conventional karyotype analysis and the in-formation about the treatment responses were also used for assessing. RESULTS: 22 of the 40 AML cases were found to carry 7 types of cytogenetic abnormalities by multiprobe FISH panel including AML1/ETO transfusion gene, PML-RARa transfusion gene, MLL breakapart, P53 deletion, Del (5q), Del7q and trisomy 8. However conventional karyotype analysis only discovered 11 cases with the corresponding cytogenetic abnormalities, the positive ratio was 57.5% in multiprobe FISH panel higher than that in karyotype analysis (27.50%). Patiens with AML1/ETO or PML-RARa transfusion gene are easily to reach CR in the first induction chemotherapy, while the Del(7q), MLL breakapart, complex cytogenetic abnormalities may indicate poor prognosis. CONCLUSION: Mutiprobe FISH panel is more rapid, accurate and effective for detecting the common cytogenetic abnormalities in AML, compared with the conventional karyotype analysis and common FISH analysis.

Analysis of efficacy and cost-effectiveness of high-dose arabinoside versus daunorubicin chemotherapy in older adult patients with acute myeloid leukemia by cytogenetic risk profile: retrospective review from China.

High-dose arabinoside (HiDAC) and daunorubicin (DNR)-based chemotherapy are the primary consolidation treatment options for older adults (50-60 years old) with acute myeloid leukemia in China. We analyzed the event-free survival (EFS) and hospital treatment charges of older adult patients with different cytogenetic risk profiles. In patients with a better/intermediate risk profile, the average total treatment cost of HiDAC was similar to that of DNR (P = 0.11). A 5-year follow-up of patients with better/intermediate cytogenetic risk profiles revealed that the median EFS of patients who received HiDAC was significantly longer than for patients who received the DNR-based regimen (27 vs. 20 months, P = 0.03). Average cost per year of life saved was 18,746.84 USD for HiDAC, compared to 32,733.37 USD for DNR. In contrast, for patients with a poor cytogenetic risk profile, the average total treatment cost for HiDAC was higher than for DNR (P < 0.005). In addition, the median EFS in the HiDAC protocol group was significantly lower than in the DNR group (11 vs. 20 months, P = 0.003). Meanwhile, in this risk group, the average cost per year of life saved was 103,237.70 USD compared to 32,277.93 USD, respectively, in the HiDAC and DNR regimens. We conclude that HiDAC is a more efficacious and cost-effective consolidation treatment regimen in the better/intermediate risk group, while the DNR-based regimen is more cost-effective in the poor risk group.

[Clinical significance of CRKL protein phosphorylation level in the treatment of chronic myeloid leukemia with imatinib].

OBJECTIVE: To investigate the adaptor protein CRKL phosphorylation level (p-CRKL) and its significance in chronic myeloid leukemia (CML) treated with imatinib. METHODS: ABL kinase domain was amplified by nested RT-PCR, domain point mutations analysis by direct sequencing, BCR-ABL mRNA level by real time-PCR, and p-CRKL level by flow cytometry in 52 bone marrow samples from 35 CML patients, and the relationship of p-CRKL level with ABL kinase domain mutation and with BCR-ABL mRNA level was analyzed. RESULTS: In the 15 imatinib-resistant patients, ABL domain point mutations were detected in 6 with 4 types of nucleotide substitutions: T315I (n = 3), Y253H (n = 1), E255K and F317L. The incidence of mutations in disease chronic phase (CP), accelerated phase (AP) and blast phase (BP) was 25.00%, 40.00% and 30.00%, respectively. The BCR-ABL mRNA level in newly diagnosed CML was higher than that in imatinib-responded patients (P = 0.01); and so did in imatinib-resistant patients than in imatinib-effective patients (P = 0.03). The level of BCR-ABL mRNA was not significantly different between newly diagnosed CML and imatinib-resistant patients. p-CRKL%, MFI showed a high degree of phosphorylation in newly diagnosed CML and imatinib-resistant patients (P = 5.130; P = 3.178). The level of p-CRKL % and MFI in newly diagnosed group was higher than that in imatinib responded group (P = 0.000; P = 0.01) and also higher in imatinib-effective group than in imatinib-resistant group (P = 0.000; P = 0.02). There was a positive correlation between the level of BCR-ABL expression and p-CRKL% (and the MFI of p-CRKL) (P < 0.05). CONCLUSION: It seems that p-CRKL detection might be helpful in predicting imatinib treatment outcomes.

[Dual-color/dual-fusion interphase fluorescence in situ hybridization probe for monitoring tumor load during imatinib therapy for chronic myeloid leukemia].

OBJECTIVE: To evaluate the sensitivity and specificity of dual-color and dual-fusion interphase fluorescence in situ hybridization (D-FISH) in determining the tumor load in patients with chronic myeloid leukemia (CML) receiving imatinib therapy. METHODS: The BCR-ABL fusion gene was detected by FISH in 24 cases of chronic myeloid leukemia treated with imatinib. The sensitivity and specificity of D-FISH were compared with those of single-fusion FISH (S-FISH) and RT-PCR. RESULTS: D-FISH was more sensitive and specific than S-FISH. In normal control subjects, the cutoff rates of D-FISH and S-FISH were 0.73% and 6.24%, respectively, showing a significant difference between them. In 24 CML cases receiving imatinib treatment, the positivity rates of S-FISH and D-FISH were 7/24 (29.2%) and 13/24 (54.2%), respectively. The results of D-FISH had a high correlation to that of RT-PCR. CONCLUSION: With lower false positive and false negative results, D-FISH can be used as a sensitive and specific method for monitoring the changes of the tumor load in CML patients during imatinib treatment.

[Preliminary study of proteins related to blast crisis in chronic myeloid leukemia].

OBJECTIVE: To identify and compare the expression profiles of differential proteins between chronic phase and blast crisis in chronic myeloid leukemia (CML) by proteomic analysis, and screen the proteins related to blast crisis. METHODS: The total cellular proteins from the bone marrow cells at chronic phase (CP) and blast crisis (BC) in CML were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and analyzed with ImageMaster 5.0 software to screen the differential protein spots. Differential protein spots were identified by mass spectrometry for peptide mass fingerprint in combination with database searching from SWISS-PROT. Then 3 protein spots were selected to verify at protein and mRNA levels by Western blot and semi-quantitative RT-PCR, separately. RESULTS: Comparing gel pages from CML-CP and CML-BC, the expression of 13 protein spots decreased and 25 protein spots increased significantly in CML-BC. Twenty differential protein spots were identified by mass spectrometry and 15 were successfully determined. The results of Western blotting were similar to those of 2-DE and showed a high expression of hnRNPK, annexin A1 and RhoA. Semi-quantitative RT-PCR analysis showed that there was no correlation between the protein expression changes and mRNA levels of hnRNPK, annexin A1 and RhoA. CONCLUSION: A group of proteins associated with blast crisis are obtained and the results may provide clues for further research to elucidate the role of these proteins in CML-BC carcinogenesis and to develop potential associated biomarkers.

[CD56 and CD11b antigen expressions in patients with acute monocytic leukemia and the clinical implications].

OBJECTIVE: To investigate the expressions of cell surface differentiation antigen CD56 and CD11b antigen in acute monocytic leukemic (AML-M(5)) cells and their clinical significance. METHODS: A total of 113 cases of de nove adult AML-M(5) were examined genetically and immunologically using G-banding technique, interphase fluorescence in situ hybridization (I-FISH) and flow cytometry immunophenotyping, and the results were analyzed in relation to their clinical data. RESULTS: Of the 113 cases, the expression rates of CD56 and CD11b was 28.32% and 73.45%, respectively. The CD56(+) patients had high CD11b expression, and the expression levels of CD11b and CD56 were positively correlated (P<0.05). The incidence of karyotypic abnormalities was 48.57% (55 cases) in these patients, including 25 (22.12%) with 11q23 aberrations. Twenty-five cases were positive for MLL gene abnormalities as found by I-FISH analysis. Compared with the patients positive for both CD56 and CD11b, those negative for both CD56 and CD11b showed increased peripheral blood white blood cell (WBC) count and also increased blast and progenitor cells in the bone marrow (P<0.05); the former patients often had karyotypic abnormalities, commonly involving 11q23 aberrations (P<0.05), whereas the latter patients presented more likely with extramedullary infiltration and refractory leukemia (P<0.01) with lowered complete remission rate and shortened median survival time (P<0.01). CD56-positive patients were more likely to have karyotypic abnormalities and refractory leukemia than CD11b-postive patients (P<0.05), but the peripheral blood WBC counts, bone marrow blast and progenitor cells, extramedullary infiltration, complete remission rate or median survival time showed no significant differences between them (P>0.05). CONCLUSION: AML-M(5) patients with CD56 positivity and high expression of CD11b often have aberrant karyotypes, commonly involving 11q23/MLL gene abnormality. These patients frequently develop extramedullary infiltration and refractory leukemia often with poor prognosis.

[Clinical characteristics and outcomes of 59 patients with acute lymphoblastic leukemia positive for BCR/ABL].

OBJECTIVE: To study the clinical characteristics and outcomes of BCR/ABL-positive acute lymphoblastic leukemia (BCR/ABL360888725-ALL) and screen the prognostic factors for BCR/ABL360888725-ALL. METHODS: From January 2001 to May 2008, 59 patients (median age of 32 years ranging from 3 to 69 years) with the diagnosis of BCR/ABL360888725-ALL by fluorescence in situ hybridization received induction chemotherapy with VDLP-/+Ara-C regimen. The patients who failed to respond to the chemotherapy received subsequent consolidation chemotherapy with imatinib (400-800 mg/day) (17 cases) or allogeneic hematopoietic stem cell transplantation (allo-HSCT) (16 cases). RESULTS: Of the 59 patients, 32 (58.3%) achieved complete remission (CR) after the first induction cycle. In patients with peripheral white blood cell (WBC) count <30=10(9)/L, 30-99.9(9)/L and > or =100(9)/L, the CR rates were 75.0% (18/24), 56.3% (9/15) and 26.3% (5/19) (P=0.006), and the overall survival probability of 2 years ( OSs of 2-yrs) was 24.7%, 22.5% and 21.1%, respectively (P=0.180). According to the FAB classification, 56 cases were divided into L1, L2 and biphenotypic acute leukemia (BAL) subgroups, and their CR rates were 66.7% (6/9), 63.2% (24/38) and 22.2% (2/9) (P=0.029), with OSs of 2-yrs of 22.2%, 27.0% and 22.0%, respectively (P=0.623). In terms of immunophenotype grouping by EGIL, the patients with ALL, myeloid antigen-positive ALL and BAL had CR rates of 61.1% (11/18), 60.6% (20/33) and 12.5% (1/8) (P=0.039), and the OSs of 2-yrs of 22.7%, 21.0% and 18.8%, respectively (P=0.643). In 55 patients with known karyotype, the CR rates were 71.4%(5/7), 70.8% (17/24) and 37.5% (9/24) in normal, sole t(9;22) abnormality, t(9;22) with additional abnormalities groups (P=0.046), with the OSs of 2-yrs of 42.9%, 34.0% and 7.3%, respectively (P=0.000). The patients complicated by septicemia had significantly lower OSs of 2-yrs than those without septicemia (0% vs 38.8%, P=0.005). The OSs of 2-yrs were significantly higher in patients with consolidation chemotherapy with imatinib than those without (48.0% vs 11.2%, P=0.001), and allo-HSCT was associated with significantly higher OSs of 2-yrs than exclusive chemotherapy (54.2% and 8.5%, P=0.000). CONCLUSION: BCR/ABL360888725-ALL with WBC> or =100 x 10(9)/L, presence of BAL diagnosed by FAB or FACM, t(9;22) with additional chromosome abnormalities all adversely affect the treatment results, and additional chromosome abnormalities and septicemia are associated with lower OSs of 2-yrs. Imatinib treatment and allo-HSCT can both improve the OSs of 2-yrs of the patients with BCR/ABL(+)-ALL.

[Detection of ABL kinase domain point mutations in chronic myeloid leukemia patients receiving imatinib treatment].

OBJECTIVE: To analyze the frequency and clinical significance of ABL tyrosine kinase point mutations in chronic myeloid leukemia (CML) patients receiving imatinib treatment. METHODS: Nested reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on 40 bone marrow samples from 23 patients to amplify the ABL kinase domain, followed by direct sequencing and sequence homologous analysis. RESULTS: In the 23 patients analyzed, the ABL domain point mutations was detected in 7 patients who presented with 5 types of nucleotide changes, namely T315I(n=3), Y253H, E255K, F317L and G321W. The incidence of mutations in chronic phase (CP), accelerated phase (AP) and blast phase (BP) was 25.00%, 40.00% and 30.00%, respectively. For 6 of the 7 patients with mutations who were resistant to imatinib before sequencing, the daily drug dose had been increased to 600-800 mg daily for poor response to 400 mg/day imatinib. During the follow-up for 3-6 months, only the patient with F317L achieved major cytogenetic response (MCR), and the patient with Y253H and 1 of the 3 with T315I progressed to BP. The newly diagnosed patient with G321W IN cp achieved a complete hematologic remission and had a significant decrease of the proportion of BCR-ABL-positive cells. CONCLUSIONS: ABL kinase point mutation is an important mechanism of imatinib resistance. The type of mutations is associated with the level of resistance to imatinib, and detection of ABL kinase point mutations by direct sequencing may help estimate the prognosis and plan for therapeutic strategy adjustment.

[Two-dimensional polyacrylamide gel electrophoresis analysis of apoptosis in K562 cells induced by harringtonine].

BACKGROUND & OBJECTIVE: Harringtonine (HT),an anti-tumor drug,has been widely used to treat acute or chronic myeloid leukemia,and obtained satisfactory effects. Studies showed that the anti-tumor activity of HT is related with the apoptosis-inducing effect,but the molecular mechanisms remain unclear. This study was to analyze the protein maps contributed to the apoptosis in K562 cells induced by HT,and to screen the apoptosis-associated proteins. METHODS: Flow cytometry was used to distinguish K562 cells of early apoptosis stage from those of late apoptosis stage through Annexin V and PI staining. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to separate and compare the HT-induced apoptotic K562 cells and control K562 cells. RESULTS: When K562 cells were treated with 10 microg/ml HT for 5,a nd 24 hours,percentages of early apoptotic cells (Annexin V+/PI-) were 28.3%, and 18.1%(P< 0.01), while percentages of late apoptotic cells (Annexin V+/PI+) were 9.1%,and 20.2% (P< 0.01), respectively. Statistical analysis showed (1300+/-50) protein spots were resolved with a match rate of (88.3+/-2.0)% in control K562 cells. Ten protein spots in late apoptotic cells displayed changes in expression after induction of HT for 24 hours (P< 0.01), of which 8 showed higher expression, 1 decreased,and 1 only expressed in control k562 cells. CONCLUSION: These proteins may be apoptosis-associated proteins of K562 cells induced by HT.

[Proteome analysis of apoptotic K562 cells induced by harringtonine].

OBJECTIVE: To screen and identify apoptosis related proteins and explore the mechanism of harringtonine (HT)-induced K562 cells apoptosis. METHODS: Flow cytometry was used to distinguish K562 cells in the earlier stage of apoptosis from those in the later stage of apoptosis by annexin V and PI staining. Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) coupled with computer image analysis was used to detect the changes in protein expression in the two stages of apoptosis. Proteins were identified by peptide mass fingerprint in combination with database searching. RESULTS: K562 cells treated with HT for 5 and 24 hours were in the early and later stages of apoptosis respectively. Statistical analysis showed 3 spots disappeared, 7 spots with decreased intensity and 10 spots with increased intensity in the 24 h HT induced apoptotic cells as compared with that in 5 h HT induced ones. Ten spots were selected on the basis of the intensity and the significant changes in abundance. Among them, 5 apoptosis related proteins were successfully identified by MALDI-TOF: keratin 9, BTF3, TrpRS, RS and prohibitin. CONCLUSIONS: Up-regulation of TrpRS, RS, prohibitin and down-regulation of BTF3 were involved in inhibition of transcription and protein synthesis in the apoptotic K562 cells induced by HT, whereas up-regulation of keratin 9 was related to apoptosis resistance.

[Clinical efficacy and side effects of STI571 in treatment of patients with chronic myeloid leukemia].

BACKGROUND & OBJECTIVE: The aberrant regulation of the protein tyrosine kinase (PTK) activity of P210(BCR-ABL), which is the protein product of Bcr-Abl fusion gene leads to the pathogenesis of chronic myeloid leukemia (CML). Though STI571 can inhibit specifically the PTK activity of P210(Bcr-Abl) and greatly improve the clinic curative effect on CML in chronic phase, its effect on CML in accelerated phase and blast crisis is not clear. In this article, we attempted to analyze the clinic efficacy and side effect of STI571 treatment on CML patients in different phase. In addition, we analyzed the potential mechanism of STI571 resistance in accelerated/blast crisis CML with genetic methods. METHODS: A total of 22 cases of CML, 14 cases male and 8 female, 6 cases in chronic phase and 16 cases in accelerated/blast crisis phase, were treated with STI571. According to the efficacy standard, the hematological and cytogenetic response of 22 cases CML were analyzed, by determining the positive rate of Ph chromosome in bone marrow from the patients treated with STI571 for 3 months. Furthermore, the karyotype evolution of those patients showing STI571 resistance was analyzed. At the same time, the side effects and adverse events of STI571 treatment were evaluated. RESULTS: 6/6(100%) cases of CML patients in chronic phase acquired hematological CR and cytogenetic response. 4/16(25%) cases in accelerated phase or blast crisis acquired hematological CR and 8/16(50%) cases acquired cytogenetic response. 3 CML patients in blast crisis showed secondary STI571 resistance. The karyotype analysis shows 2 with 2 Ph chromosome and other additional abnormality. I/II grade non-hematological toxicity was observed in all the patients, including edema (77.3%), side effects of digestive system (36.4%) and myalgia (22.7%) et al. Severe hematological toxicities includes:(1)III/IV grade neutropenia (9 cases):1/6 cases of CML patients in chronic phase, 8/16 cases in accelerated phase or blast crisis; (2)III/IV grade thrombocytopenia (6 cases): 6/16 cases in accelerated phase or blast crisis. The percentage of III/IV grade neutropenia/thrombocytopenia in chronic phase and accelerated/blast crisis phase was compared and no significant statistical difference was observed. CONCLUSIONS: Hematological and cytogenetic responses of different degrees can be acquired in CML-CP and CML-AP/BC patients treated with STI571 and showing statistic difference. STI571 improves the clinic curative effect greatly on CML in chronic phase. CML patients in blast crisis have secondary STI571 resistance with novel 2 Ph chromosome and other additional abnormality, it furnishes the evidence of the gene changes. The slightness of non-hematological toxicity of STI571 in the treatment of chronic myeloid leukemia suggests this drug is relatively safe. Severe hematological toxicities, such as III/IV neutropenia and thrombocytopenia, are more common in accelerated/blast crisis than in chronic phase.