[Clinical study of induction chemotherapy followed by allogeneic hematopoietic stem cell transplantation in the treatment of FLT3-ITD(+) acute myeloid leukemia with normal karyotype].

Objective: To assess the efficacy of induction chemotherapy followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of FLT3-ITD(+) acute myeloid leukemia (AML) with normal karyotype. Methods: The clinical data of FLT3-ITD(+) AML patients with normal karyotype in the First Affiliated Hospital of Nanjing Medical University from Jan 2018 to March 2021 were retrospectively analyzed. Results: The study included 49 patients with FLT3-ITD(+)AML, 31 males, and 18 females, with a median age of 46 (16-59) years old. All patients received induction chemotherapy, and 24 patients received sequential allo-HSCT (transplantation group) . The median follow-up time was 465 days, the one-year overall survival (OS) from diagnosis was (70.0 ± 7.4) %, and one-year disease-free survival (DFS) was (70.3±7.4) %. The one-year OS was significantly different between the transplantation group and the non-transplantation group [ (85.2 ± 7.9) % vs (52.6 ± 12.3) %, P=0.049]. but one-year DFS [ (84.7 ± 8.1) % vs (55.2 ± 11.9) %, P=0.061] was not. No significance was found in one-year OS between patients with low-frequency and high-frequency FLT3-ITD(+) (P>0.05) . There were 12 patients with high-frequency FLT3-ITD(+) in the transplantation and the non-transplantation groups, respectively. The one-year OS [ (68.8 ± 15.7) % in the transplantation group vs (26.2 ± 15.3) % in the non-transplantation group, P=0.027] and one-year DFS [ (45.5 ± 21.3) % in the transplantation group vs (27.8±15.8) % in the non-transplantation group, P=0.032] were significantly different between the two groups. Conclusion: Induction chemotherapy followed by allo-HSCT can enhance the prognosis of FLT3-ITD(+) patients, particularly those with FLT3-ITD high-frequency mutation.

[The role and mechanism of lncRNA C9ORF139 targeting miR-24-3P/TAOK1 in regulating the proliferation of acute myeloid leukemia cells].

Objective: To investigate the role and mechanism of long non-coding RNA (lncRNA) C9ORF139 targeting micro RNA(miR)-24-3P/TAOK1 in regulating the proliferation of acute myeloid leukemia (AML) cells. Methods: AML cells HL-60 and THP-1 were purchased from the Chinese Academy of Sciences and divided into 4 groups:group A was negative control group (siNC group), group B was interference C9ORF139 group (siC9ORF139 group), group C was siC9ORF139+miR-24-3p inhibitor group, and group D was miR-24-3P+TAOK1 overexpression group (oe-TAOK1 group). Real-time fluorescence quantitative reverse transcription PCR was used to detect the expression levels of AML cell lines of HL-60 and THP-1 in four groups. Cell Counting Kit-8 assay was performed to measure cell proliferation. Flow cytometry was applied to analyze cell apoptosis. Transwell test was applied to detect cell migration and invasion ability. Western blot was used to detect p-serine/threonine kinase (p-raf) and p-mitogen activation proteinkinase (p-MEK), p-extracellular regulatory protein kinase (p-ERK) expression. The luciferase reporter gene plasmid was constructed to verify the binding ability of C9ORF139,miR-24-3P and TAOK1.Nude mice were inoculated with subcutaneous tumor cells of HL-60 (group A) and HL-60 (group B). Results: After the C9ORF139 gene was knocked down and cultured for 120 h, The cell proliferation ability (0.62±0.02, 0.82±0.02), migration ability (0.22±0.03, 0.05±0.01), invasion ability (0.20±0.02, 0.13±0.03) of group B were all lower than that of group A (1.30±0.02, 1.83±0.07; 0.99±0.02, 0.99±0.02; 1.00±0.01, 1.00±0.01) (all P<0.05). When co-transfected with miR-24-3 inhibitor, cell proliferation ability, migration ability and invasion ability were all higher in group B (all P<0.05). When co-transfected with miR-24-3P and oe-TAOK1 plasmid, cell proliferation ability, migration ability and invasion ability were all higher than group B (all P<0.05).When the C9ORF139 gene in the cells was knocked down, the apoptosis level of group B (28.56±8.07, 17.74±1.91) were higher than those of group A (0.31±0.27, 2.49±0.33)(all P<0.05); when co-transfected with miR-24-3P inhibitor, the apoptosis level (2.34±0.09, 3.06±0.06) were lower than those in group B (all P<0.05); when co-transfected with miR-24-3P and oe-TAOK1 in the plasmid group, the apoptosis level (2.16±1.29, 4.80±0.37) were also lower than those of group B (all P<0.05). In HL-60 and THP-1 cells, when C9ORF139 was not mutated, the luciferase activity of miR-24-3P group was lower than that of the miR-NC group (P<0.05). When the binding site with miR-24-3p in C9ORF139 sequence was mutated, the luciferase activity in miR-24-3p group was equivalent to that in miR-NC group (P>0.05).When TAOK1 was not mutated; the luciferase activity of miR-24-3P group was lower than that of group A (P<0.05). When the binding site with miR-24-3p in TAOK1 sequence was mutated, the luciferase activity in miR-24-3p group was equivalent to that in miR-NC group (P>0.05).When the C9ORF139 gene in HL-60 cells was knocked down and cultured for 72 h, the phosphorylation expression levels of Raf, MEK and ERK molecules in group B were significantly lower than those in group A (all P<0.05). By day 14, the tumor volume in the group A was greater than the tumor cell volume in the group B [(284.49±57.61) vs (125.70±18.64) mm3, P=0.017]. The tumor weight of HL-60 in group A was heavier than that of group B [(847.80±159.36) vs (408.40±113.16) mg, P=0.001]. Conclusions: LncRNA C9ORF139 regulates TAOK1 by sponging miR-24-3P to promote the proliferation, invasion and migration of acute myeloid leukemiacell.In vivo experiments have confirmed that the expression of C9ORF139 can promote the growth of subcutaneous tumors in AML nude mice.

[Estradiol regulates the expression of plasma membrane Ca2+-ATPase isoform 2 in inner ear of rats].

Objective: To observe the effects of estradiol on expression of plasma membrane Ca2+-ATPase isoform 2 in inner ear of rats. Methods: Twenty-five Three-months-old female Sprague-Dawley rats were randomly and equally divided into five groups by random number table mathod,with five rats in each group. Animals in Sham group were sham-operated while others were bilateral ovariactmized. One month after modeling, the OVX groups were supplemented with estradiol (E2 group), progesterone (P group), estradiol and progesterone (E2+P group)and vehicle sesame oil (Veh group), while the Sham operation group (Sham group) was supplemented with vehicle sesame oil.All rats were sacrificed and otocysts were obtained immediately. Enzyme-linked immunosorbent assay was used to detect the changes in serum estradiol and progesterone levels of each group of rats before operation, before treatment and before sacrifice. Western blot and quantitative real-time polymerase chain reaction were used to detect the expression of total PMCA2 protein and mRNA in the inner ear of each group. Results: There was no significant difference in serum estradiol and progesterone levels among the five groups before operated(P>0.05). Before treatment, the serum estradiol and progesterone levels of rats in each group were significantly lower than those in Sham group (P<0.05). The serum estradiol level in E2 group and E2+P group was not significantly different from that in Sham group (P>0.05), while the serum estradiol level in P group and Veh group was significantly different from that in Sham group (P<0.05). The level of progesterone in P group and E2+P group was higher than that in Sham group (P<0.05), while the level of progesterone in Veh group and E2 group was lower than that in Sham group (P<0.05). Protein and mRNA expression of PMCA2 in P and Veh groups were significantly decreased compared with that of Sham group (P<0.05) while the expression levels underwent no significantly change in E2 and E2+P groups (P<0.05). Conclusion: The decrease of serum estradiol level can reduce the expression of otolith regulatory protein PMCA2 in rats, and then affect otolith metabolism, which may be an important link of estrogen affecting otolith metabolism.

[Clinical features and prognosis of low triiodothyronine syndrome in patients with acute myeloid leukemia].

Objective: To investigate the clinical features and prognosis of low triiodothyronine syndrome (LT3S) in patients with acute myeloid leukemia (AML) . Methods: A total of two 236 patients with AML who presented at the Jiangsu Provincial Hospital between January 2013 and December 2019 were included, and their data were retrospectively reviewed. The patients were divided into two groups, including the LT3S group and the non-LT3S group, according to their serum thyroxine level. The clinical characteristics and prognosis of the two groups were compared. Results: Among the 236 patients, 62 (26.3%) patients had LT3S. Serum-free T3 level was positively correlated with albumin (r=0.443, P<0.001) and hemoglobin (r=0.187, P=0.005) levels and negatively correlated with C-reactive protein (r=-0.406, P<0.001) and lactate dehydrogenase (r=-0.274, P<0.001) levels. The overall survival (OS) (7.5 months vs 29.9 months, P<0.001) and progression-free survival (PFS) (2.0 months vs 24.0 months, P<0.001) were significantly shortened in the LT3S group compared with the non-LT3S group. After propensity score matching, the OS (9.6 months vs 30.4 months, P=0.010) and PFS (3.0 months vs 30.0 months, P=0.014) were still significantly reduced in the LT3S group compared with the non-LT3S group. Therefore, LT3S was an independent risk factor for OS (HR=2.553, 95% CI 1.666-3.912, P<0.001) and PFS (HR=1.701, 95% CI 1.114-2.597, P=0.014) in patients with AML. Subgroup analysis suggested that patients with LT3S had a worse prognosis in patients with AML who were obese, fragile, or treated with standard chemotherapy. Conclusions: The occurrence of LT3S reflects the poor clinical status and prognosis of patients with AML.

[Active screening of intestinal carbapenem-resistant Enterobacteriaceae in high-risk patients admitted to the hematology wards and its effect evaluation].

Objective: To evaluate the effect of intestinal carbapenem-resistant Enterobacteriaceae (CRE) active screening combined with enhanced intervention in the prevention and control of nosocomial infection in patients admitted to the hematological ward. Methods: Patients who were admitted to the Department of Hematology in a tertiary-care general hospital from March 1, 2017 to December 31, 2019 and underwent chemotherapy or immunosuppressive therapy comprised the intervention group. They were screened for intestinal CRE at least thrice. From December 1, 2016 to February 28, 2017, patients who underwent chemotherapy or immunosuppressive therapy without active intestinal CRE screening in the Department of Hematology formed the control group. Both the patient groups were monitored for CRE infection in real time. The χ(2) test was used to compare the changes in the CRE infection rate and mortality in high-risk patients before and after the active screening. Results: During the intervention period, the CRE colonization rate of patients was 16.46% (66/401) ; in terms of disease distribution, the colonization rate of acute leukemia was the highest 23.03% (26/113) . Of the 66 colonized patients, 27 (40.9%) patients were identified as positive for CRE at the first screening, 15 (22.7%) were identified at the time of the second screening, and the remaining 24 (36.4%) were identified at the third or subsequent screening; Carbapenem-resistant Klebsiella pneumoniae (CRPK) strains were dominant among the pathogens, accounting for 54.55% (36/66) . During the active screening period, the CRE infection rate (2.49%) and mortality rate (50.00%) of high-risk patients were significantly lower than those of the controls (11.30% and 69.23%, respectively) . The pathogens of 10 CRE infection patients during the intervention period were exactly the same as the previous active screening pathogens, and the coincidence rate was 100%. Conclusion: The CRE colonization rate was the highest in patients with acute leukemia who were admitted in the hematology wards. CRPK is the main pathogen of CRE colonization, infection, and death. Increasing the frequency of screening can significantly raise the positive rate of screening, Active screening can effectively reduce the incidence and subsequent mortality of CRE in high-risk patients admitted in the hematological wards. High coincidence rate between CRE screening positive pathogens and subsequent CRE infection pathogens. Intestinal CRE screening can serve as an indicator of CRE bloodstream infection in patients with hematological diseases as well as provide information for antibiotics therapy.

Serum oxidative stress, inflammatory response and platelet activation in patients with vascular vertigo.

The aim of this study was to investigate the correlation and clinical significance of oxidative stress and inflammatory response in vascular vertigo (VV). The subjects were divided into three groups: vascular vertigo (group A), non-vascular vertigo (group B) and controls (group C). The serum levels of IL-6 (interleukins-6), SOD (superoxide dismutase), MDA (malondialdehyde) and TNF-α (tumor necrosis factor-α) and CD62P (also called P-Selectin) activation rates were determined and compared among the three groups. The levels of IL-6, TNF-α, MDA and CD62P in group A were significantly higher than those of group B and group C (P less than 0.05). The SOD level of group A was lower than that of group B and group C (P less than 0.05). There was no significant difference between groups B and C in IL-6, TNF- αMDA, SOD and CD62P (P>0.05). In patients with vascular vertigo, TNF-α levels had a weak linear correlation with those of low-density lipoprotein (P = 0.025, r = 0.312). There was no linear correlation between TNF-α and SOD in patients with VV and non-VV. The occurrence of inflammatory reaction and oxidative stress may cause abnormal lipid metabolism in the body and promote the occurrence of VV, and platelet activation may be involved in its formation.

[The prognostic significance of plasma Epstein-Barr virus DNA in patients with diffuse large B cell lymphoma].

To retrospectively analyze the prognostic significance of plasma Epstein-Barr virus (EBV) DNA in 122 patients with diffuse large B cell lymphoma (DLBCL). Plasma EBV DNA positivity was related to advanced disease stage (P=0.030), B symptoms (P=0.004) and elevated serum lactic dehydrogenase (LDH) (P=0.001). Furthermore, univariate analysis indicated that plasma EBV DNA level was associated with worse overall survival (OS) (HR=0.223, 95%CI 0.096-0.518, P<0.001) and worse progression free survival (PFS) (HR=4.417, 95%CI 1.911-10.208, P<0.001), whereas multivariate analysis showed plasma EBV DNA as a probable independent prognostic factor of clinical outcome(HR=0.409, 95%CI 0.166-1.008, P=0.052).

[Misdiagnosis and associated costs of benign paroxysmal positional vertigo].

Objective: The aims of this study were to investigate the misdiagnosis of benign paroxysmal positional vertigo (BPPV) and to estimate the associated costs. Methods: During October 2015 to December 2015, eighty patients were diagnosed with BPPV in the outpatient dizziness clinic of Shanghai Changzheng Hospital and the clinical data of all the 80 patients were collected including the demographic and clinical characteristics, the history of diagnosis, inappropriate diagnostic tests, costs of the medical tests, transportation and accommodation. All the data were investigated to estimate the misdiagnosis of benign paroxysmal positional vertigo and the associated costs in Shanghai, China. Results: This study showed that the misdiagnosis rate of BPPV was 60.0% (48/80) and the common inappropriate diagnostic tests for BPPV included Cranial CT and MRI test, cervical MRI, cervical and cerebrovascular investigations et al. There was no significant difference between the misdiagnosis patients (48) and patients without misdiagnosis (32) in gender, age, duration of symptom, involved canal and type of BPPV.Complications were significantly more frequent in the misdiagnosis group than for those without[81.3%(39 /48) vs 34.4%(11 /32)]. The estimated costs for each misdiagnosed individual were 8 502.98 China Yuan (CNY) and one-year economic burden associated with the misdiagnosis of BPPV in Shanghai was 13.184 7-78.862 1 million CNY. Conclusions: Our study suggests that the misdiagnosis rate of BPPV is high and the financial impact on patients and society with this disease is huge. The cost-effective Dix-Hallpike or supine roll test maneuver should be used before applying other expensive medical tests in order to minimize misdiagnosis and the waste of health care resources.

Immune- and ribosome-related genes were associated with systemic vasculitis.

This study aimed to investigate the molecular mechanism of systemic vasculitis via bioinformatics analysis. Gene express profile of E-GEOD-16945 (13 Takayasu arteritis samples and 13 control samples) was downloaded from European Bioinformatics Institute (EBI) database. Differentially expressed genes (DEGs) were screened between Takayasu arteritis and normal controls (|log FC| > 1). Basic local alignment search tool (BLASTX) was used for the Clusters of Orthologous Groups (COG) classification of DEGs. Gene ontology analysis was performed for the DEGs (P < 0.05). A gene expression network was built with DEGs. Mcode in Cytoscape software was used to extract modules from the network (degree ≥ 2, K-core ≥ 2 and adjusted P-value < 0.05) followed by pathway analysis using GenMAPP (false discovery rate < 0.05). A total of 747 DEGs were identified. There were 16 significant GO function terms enriched with DEGs, of which immune and defence response was the most significant GO term. Totally, three modules were extracted from gene expression network, including one module constituted with upregulated genes and two modules constituted with downregulated genes. Furthermore, human leucocyte antigen (HLA)-DRB1, HLA-DPA1, HLA-DPB1, HLA-DOA and HLA-DRA in the downregulated modules were significantly linked to immune-related pathways (intestinal immune network for IgA production and systemic lupus erythematosus pathways), while ribosomal protein L 31 (RPL31), RPS3A and RPL9 in the upregulated module were enriched in ribosome pathway. The immune-related pathways, ribosome pathway, immune-related genes including (HLA-DRB1, HLA-DPA1, HLA-DPB1, HLA-DOA and HLA-DRA) and ribosome-related genes (RPL31, RPS3A and RPL9) might be involved in systemic vasculitis.

Molecular cytogenetic aberrations in patients with multiple myeloma studied by interphase fluorescence in situ hybridization.

BACKGROUND: Multiple myeloma (MM) is an incurable hematological disorder characterized by the accumulation of malignant plasma cells within the bone marrow (BM). The clinical heterogeneity of MM is dictated by the cytogenetic aberrations present in the clonal plasma cells (PCs). Cytogenetic studies in MM are hampered by the hypoproliferative nature of plasma cells in MM. Therefore, fluorescence in situ hybridization (FISH) analysis combined with magnetic-activated cell sorting (MACS) is an attractive alternative for evaluation of numerical and structural chromosomal changes in MM. METHODS: Interphase FISH studies with three different specific probes for the regions containing 13q14.3(D13S319), 14q32(IGHC/IGHV) and 1q12(CEP1) were performed in 48 MM patients. Interphase FISH studies with LSI IGH/CCND1, LSI IGH/FGFR3, and LSI IGH/MAF probes were used to detect t(11;14)(q13;q32), t(4;14)(p16;q32), and t(14;16)(q32;q23) in patients with 14q32 rearrangement. RESULTS: Molecular cytogenetic aberrations were found in 40 (83.3%) of the 48 MM patients. 13 patients (27.1%) simultaneously had 13q deletion/monosomy 13[del(13q14)], illegitimate IGH rearrangement and chromosome 1 abnormality. Del (13q14) was detected in 21 cases (43.7%), and illegitimate IGH rearrangements in 29 (60.4%) including 6 with t (11;14) and 5 with t(4;14). None of 9 patients with illegitimate IGH rearrangements and without t(11;14) or t(4;14) we detected had t(14;16)(q32;q23). 24 of the 48 MM patients (50%) had chromosome 1 abnormalities. Among 21 patients with del (13q14), 15 patients had Amp1q12;16 had IgH rearrangements. Whereas, among 27 cases without del (13q14), 8 had Amp1q12; 13 had IgH rearrangements. There was a strong association between del(13q14) and Amp1q12 ( = 8.26, p < 0.01), and between del (13q14) and IgH rearrangement ( = 3.88, p < 0.05). CONCLUSION: 13q deletion/monosomy 13, IGH rearrangement and chromosome 1 abnormality are frequent in MM. They are not randomly distributed, but strongly interconnected. Interphase FISH technique combined with MACS using CD138-specific antibody is a highly sensitive technique at detecting molecular cytogenetic aberrations in MM.

Conventional and molecular cytogenetic features of myelodysplastic syndrome in China.

BACKGROUND: Myelodysplastic syndrome (MDS) constitutes a heterogeneous group of hematopoietic stem cell disorder characterized by peripheral blood cytopenia(s), in the presence of hypercellular bone marrow with features of ineffective hematopoiesis, and susceptibility to acute leukemia (AL). Although the precise pathogenesis of MDS remains to be clarified, cytogenetic abnormalities seem to be involved in its pathogenesis and are considered as an important factor in diagnosis and predicting clinical outcome. OBJECTIVE: To explore the cytogenetic features of Chinese patients with myelodysplastic syndrome (MDS). METHODS: Conventional cytogenetic analysis was performed in 88 MDS patients and among them, 34 cases were studied by interphase fluorescence in situ hybridization (I-FISH) with precisely chromosome 8 centromere specific DNA probe and DNA specific probes for 7q32 , 5q31. RESULTS: Of the 88 patients, 45 (51.1%) showed clonal karyotypic abnormalities by CC at diagnosis, including numerical changes (18 cases, 20.5%), structural changes (12 cases, 13.6%), and numerical and structural changes simultaneously(15 cases, 17.0%). Trisomy 8, -5/5q-, and -7/ 7q- account for 20.5%, 15.9%, and 5.7% respectively. Complex karyotypes were observed in 17 patients, the incidence being 19.3% in the whole series of cases. Among 34 MDS patients studied by I-FISH, -5/5q-, -7/7q- and trisomy 8 occurring in 4, 2 and 10 cases respectively for CC were confirmed by I-FISH. 5 cases in 30 cases who did not show -5/5q- by CC displayed this abnormality by I-FISH. 3 cases without -7/7q- by CC presented this aberration by I-FISH. 5 cases with trisomy 8 for I-FISH was not identified this change by CC. CONCLUSIONS: The frequent abnormalities are trisomy 8, -5/5q- and -7/ 7q-. FISH is very useful in detecting these alterations in MDS and it is an important complement to CC.

Multiphoton route to ZnO nanowire lasers.

With intense femtosecond laser excitation, multiphoton absorption-induced stimulated emission and laser emission in ZnO bulk crystal and nanowires have been demonstrated at room temperature. UV-stimulated emission peaks appeared in both bulk crystal and nanowires when the excitation exceeded certain thresholds, and a sharp lasing peak with a linewidth of ~0.5 nm was observed from ZnO nanowires. The emission properties were attributed to the band-edge emission of the recombination of carriers excited by two- and three-photon absorption processes in the wide-bandgap semiconductor.

Intraurethral brachytherapy for prevention of recurrent urethral stricture after internal urethrotomy or transurethral resection of scar.

BACKGROUND PURPOSE: Restricture after internal urethrotomy is the major limitation to the long-term success of the procedure. The objective of this study was to evaluate the effect of intraurethral brachytherapy after internal urethrotomy or transurethral scar resection on recurrent urethral stricture. PATIENTS AND METHODS: From January 1998 to June 1999, catheter-based intraurethral brachytherapy with 192-iridium was performed in 17 patients with recurrent urethral stricture to prevent restricture after internal urethrotomy or transurethral resection of scar. The radiation was repeated within 3 days after surgery to reach a total dosage of 1000 to 1500 cGy. RESULTS: During the follow-up (range 14-27 months; mean 20 months), two patients had dysuria, including one patient with an atonic detrusor muscle. The other patient needed self-dilation. Fifteen patients presented normal voiding. The stricture recurred 3 months later in only one patient, so the restricture rate is 7%. No significant complication was observed associated with brachytherapy during the follow-up. CONCLUSION: Intraurethral brachytherapy after internal urethrotomy or transurethral resection of scar is a safe and effective treatment for recurrent urethral strictures.

[Mechanism of p53 gene mutation in the development of urologic cancer].

With DNA polymerase chain reaction-single strand conformation polymorphism assay followed by direct DNA sequencing, p53 gene mutation was examined in bladder transitional epithelial cell carcinoma, renal cell carcinoma and testicular seminoma. p53 gene mutation was found in 7 cases (35%) of bladder carcinoma and 4 cases (23.5%) of testicular seminoma. Inactivation of Rb gene and activation of ras and c-erbB-2 were also studied. The results suggest that development of urologic neoplasms is closely associated with p53 gene mutation and involves loss of expression of Rb and aberrant expression of ras and c-erbB-2.