Comprehensive Landscape of HOXA2, HOXA9, and HOXA10 as Potential Biomarkers for Predicting Progression and Prognosis in Prostate Cancer.

Prostate cancer (PCa) is recognized as a common malignancy in male patients. The homeobox A cluster (HOXA) family members have been confirmed to be implicated in the development of several types of tumors. However, the expression pattern and prognostic values of HOXA genes in PCa have not been investigated. In this study, we analyzed TCGA datasets and identified six HOXA family members which showed a dysregulated expression in PCa specimens compared with nontumor specimens. We also explored the potential mechanisms involved in the dysregulation of HOXA family members in PCa, and the results of Pearson's correlation revealed that most HOXA members were negatively related to the methylation degree. Moreover, we explored the prognostic values of HOXA family members and identified six survival-related HOXA members. Importantly, HOXA2, HOXA9, and HOXA10 were identified as critical PCa-related genes which were abnormally expressed in PCa and associated with clinical outcomes of PCa patients. Then, we explored the association between the above three genes and immune cell infiltration. We observed that the levels of HOXA2, HOXA9, and HOXA10 were associated with the levels of immune infiltration of several kinds of immune cells. Overall, our findings identified the potential values of the HOXA family for outcome prediction in PCa, which might facilitate personalized counselling and treatment in PCa.

[The Clinical Characteristics and Prognosis of Patients with Acute Myeloid Leukemia Combine Paroxysmal Nocturnal Hemoglobinuria].

OBJECTIVE: To investigate the clinical characteristics and prognosis of patients with acute myeloid leukemia(AML) combined with paroxysmal nocturnal hemoglobinuria(PNH). METHODS: The clinical data of 13 AML combined with PNH patients treated in our hospital from January 2017 to May 2019 were collected and retrospective analyzed. The complete remission(CR) rate for induction chemotherapy was analyzed. The level of PNH+ cell before and after chemotherapy were tested by Paired t test. Kaplan-Meier method and multi-factorial Cox regression model were used to analyze the influencing factors of prognosis. RESULTS: Among the 13 patients, 11 (84.6%) cases were CR after first induction chemotherapy. The median overall survival(OS) time was 17 months(0-30 months), the median progression-free survival(PFS) time was 16 months(2-26 months). There were no significant difference in the number of PNH+ cell before and after chemotherapy (P＞0.05). Multivariate Cox regression analysis showed that age，sex，the level of hemoglobin, platelet were not related to the OS of the patients(P＞0.05), the level of WBC, LDH and risk stratification at first diagnosed were related to the OS of the patients（P＜0.05）. Kaplan-Meier survival analysis showed that the OS rate of AML combined with PNH patients with leukocyte lower than 10×109/L at first diagnosed was better than that of the patients with leukocyte higher than 10×109/L (P=0.0261). The OS rate of patients with low or standard risk was better than the patients with high risk group(P=0.0010). CONCLUSION: The patients of AML combined with PNH have higher CR rate after the first induction chemotherapy. The level of WBC and LDH at first diagnosed are the factors that affecting the OS of the patients. The OS of patients with WBC lower than 10×109/L, at first diagnosed low and medium risk are better than the other patients.

Clinical Characteristics and Prognostic Analysis of Multiple Myeloma with Extramedullary Disease: A SEER-Based Study.

BACKGROUND: Extramedullary disease (EMD), an infrequent manifestation of multiple myeloma (MM), can present at diagnosis or develop during the disease course. EMD can be clinically divided into bone-related EMD (EMD-B) and soft tissue-related EMD (EMD-S). The purpose of our study is to investigate the clinical characteristics, survival outcomes, and prognostic factors of MM patients with EMD. METHODS: A total of 155 MM patients with EMD were ultimately enrolled in our study by retrieving the Surveillance, Epidemiology, and End Results (SEER) database. The Kaplan-Meier survival curves and log-rank test for overall survival (OS) and myeloma-specific survival (MSS) were conducted to compare each potential variable. Variables with a p value <0.1 in the univariate Cox regression were incorporated into the multivariate Cox model to determine the independent prognostic factors, with a hazard ratio (HR) >1 representing adverse prognostic factors. RESULTS: The median age at diagnosis was 63 years old. EMD-B occurred in 99 patients (63.90%), while EMD-S occurred in 56 cases (36.10%). Patients with EMD-S had a significant survival disadvantage in MSS (HR = 1.844, 95% CI 1.117-3.042, p = 0.017) and OS (HR = 1.853, 95% CI 1.166-2.942, p = 0.009) compared to those with EMD-B. Patients with EMD interval ≤24 months were at higher risk of death than those with EMD at diagnosis in MSS (HR = 1.885, 95% CI 1.175-3.346, p = 0.042) and in OS (HR = 1.33, 95% CI 1.119-2.529, p = 0.036). Patients with EMD interval >24 months were at a lower risk of death as opposed to those with EMD at diagnosis. CONCLUSION: Age at MM diagnosis, site of EMD, and time interval from diagnosis to EMD occurrence were independent prognostic factors in MM patients with EMD. EMD-B bore a better prognosis than EMD-S.

Genotype profiles of Helicobacter pylori from gastric biopsies and strains with antimicrobial-induced resistance.

BACKGROUND AND AIMS: The genotypic method could significantly shorten the time needed to obtain antibiotic susceptibility data for Helicobacter pylori. The aim of this study was to explore the profile of H. pylori from gastric biopsies and strains with antibiotic-induced resistance. METHODS: A total of 124 gastric biopsies were used to perform gene sequencing and to perform bacterial culture and susceptibility testing. Seven susceptible strains were selected to develop resistance to clarithromycin, levofloxacin, and metronidazole. Four susceptible strains were selected to transfer candidate mutations. The genotype profiles of these groups were analyzed by sequencing analysis. The antibiotic susceptibility of these strains was detected using the E-test method. RESULTS: Phenotypic resistance to clarithromycin, levofloxacin, and metronidazole was observed in 35.5%, 40.0%, and 79.8% strains, respectively. Point mutations in 23 S rRNA, gyrA, and rdxA genes were observed in 39.5%, 38.7%, and 86.3% of gastric biopsies, respectively. The A2143G mutation in the 23S rRNA occurs in most clarithromycin-resistant samples. The A2142C point mutation showed a higher efficacy than A2142G and A2143G for inducing clarithromycin resistance. The D91N and N87K mutations in gyrA occurs in most levofloxacin-resistant samples, and double point mutations showed a higher efficacy than single mutations for inducing levofloxacin resistance. Phenotypic resistance and mutations in rdxA lacked consistency. CONCLUSION: Genotype-based gastric biopsy analysis was reliable for determining clarithromycin and levofloxacin resistance. A2143G in 23S rRNA and N87K/D91N in the gyrA gene occurred in most resistant strains. Mutations in the rdxA gene were not good indicators of metronidazole resistance.

BRD4 inhibitor nitroxoline enhances the sensitivity of multiple myeloma cells to bortezomib in vitro and in vivo by promoting mitochondrial pathway-mediated cell apoptosis.

BACKGROUND: Multiple myeloma (MM) is the second most common hematological neoplasm. Wide administration of bortezomib significantly improves the survival of MM patients compared with conventional chemotherapy. Bromodomain-containing protein 4 (BRD4) inhibitors also have been demonstrated to retard cell proliferation and induce cellular apoptosis in various cancers. However, it is unclear whether the BRD4 inhibitor nitroxoline plus bortezomib has a synergistic anti-tumor effect on MM. METHODS: Cell viability was determined via 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell cycle and cell apoptosis were assessed via flow cytometry. Protein expression levels were determined via western blotting. The expression of apoptosis-related proteins in xenograft tissue were detected by means of immunohistochemistry. RESULTS: Treatment with nitroxoline or bortezomib suppressed cell proliferation, and caused G0/G1 phase arrest and apoptosis in H929 and RPMI8226 cells. Furthermore, nitroxoline intensified the retardation of cell proliferation, as well as further enhanced the G0/G1 phase arrest and apoptosis induced by bortezomib in H929 and RPMI8226 cells. The western blot analysis revealed that nitroxoline or bortezomib treatment markedly diminished the levels of Bcl-2 and cyclin D1, and increased the levels of p21, Bax, cleaved PARP and cleaved caspase-3. Combination of these two agents was observed to result in further marked changes on these levels compared with nitroxoline or bortezomib treatment alone. What is more, in the xenograft tumor model, combinative treatment markedly inhibited tumor growth compared with the single drug treatment. CONCLUSION: Combination of bortezomib with nitroxoline has a synergistic anti-tumor activity in MM cells and may be a novel treatment method for MM.

The Evaluation of a Two-Year Outcome of Intravitreal Conbercept versus Ranibizumab for Pathological Myopic Choroidal Neovascularization.

Aim: The aim of this study was to evaluate and compare the 24-month follow-up results of intravitreal conbercept with ranibizumab in the treatment of choroidal neovascularization (CNV), secondary to pathological myopia (PM). Methods: Fifty-nine patients' 64 eyes with pathological myopic CNV were retrospectively reviewed. Thirty-one eyes underwent conbercept treatment (group C) and 33 eyes underwent ranibizumab treatment (group R), respectively. No patients have received any treatment before. The main outcome of the best-corrected visual acuity (BCVA) uses an international standard visual acuity chart and is converted to the minimum resolution logarithm (LogMAR) visual acuity. Results: There were no significant differences between the two groups at the baseline statisticlly. At 24 months, the mean logMAR BCVA of group C increased from 0.95 ± 0.54 to 0.58 ± 0.39 (P < .001) and the mean central macular thickness (CMT) decreased from 280.97 ± 62.69 µm to 242.35 ± 90.39 µm (P = .033). The mean logMAR BCVA of group R increased from 0.86 ± 0.40 to 0.54 ± 0.28 (P < .001) and the mean CMT was reduced from 303.58 ± 61.95 µm to 251.82 ± 84.74 µm (P = .005). There was no significant difference in logMAR BCVA and CMT between the two groups (P = .962, P = .667, respectively). The mean number of injections was 3.94 ± 1.88 in group C and 4.06 ± 1.82 in group R (P = .788). During the follow-up period, no ocular complications and systemic adverse reactions were observed. Conclusion: Similar visual acuity and improved morphology were achieved in both groups. The two drugs were also found to be safe and effective in the treatment of pathological myopic CNV.

Evaluation of outer retinal tubulations in diabetic macular edema underwent anti-VEGF treatment.

AIM: To investigate the incidence and subsequent changes of outer retinal tubulations (ORTs) in diabetic macular edema (DME) underwent anti-vascular endothelial growth factor (VEGF) therapy, and to assess the possibility of ORT as a biomarker of DME severity or response to anti-VEGF therapy. METHODS: This retrospective and descriptive study included a total of 228 patients (435 eyes) with DME and treated with intravitreal anti-VEGF agents between March 2016 and January 2018. Patients were divided into 2 groups according to the presence of ORTs. High-resolution spectral-domain optical coherence tomography (SD-OCT) images acquired by vertical and horizontal scans and over consecutive visits were analyzed. The evolution of ORT over time, type of fluid and subfoveal photoreceptor integrity on OCT imaging was also assessed. RESULTS: ORTs were identified in 108 eyes of 435 eyes with an overall incidence rate of 24.83% at baseline. ORTs were prone to locate adjacent to the lesions of exudation and/or cystoid edema and possibly situated in outer nuclear layer (ONL), outer plexiform layer (OPL) and/or inner nuclear layer (INL) in eyes with DME. The formation process of ORT led to focal downward displacement of OPL and INL toward RPE near the lesion. During the follow up, 45 eyes had steady ORTs and 63 eyes had dynamic variants in ORTs, including disappearance, reappearance, collapse, diminution, and enlargement. There were higher proportion of closed ORTs and fewer proportion of forming ORTs in eyes with steady ORTs, which showed a statistically significance when compared with eyes with variant ORTs (P=0.006, P=0.017, respectively). The eyes without ORTs had significantly better final best corrected visual acuity (BCVA) and more BCVA change than those eyes with ORTs in DME patients after anti-VEGF therapy (P=0.023, P=0.009, respectively). The disruption of subfoveal photoreceptor integrity in eyes with ORTs was more serious than that in eyes without ORTs (P=0.013). The proportion of stable vision in eyes with ORTs was significantly higher than that in eyes without ORTs, showing statistical significance (P=0.016). ORTs were associated with worse visual prognosis due to damage of the subfoveal photoreceptor integrity. CONCLUSION: ORTs have a high incidence and changes over time in DME with anti-VEGF treatment and may be located at various retinal layers. Persistent ORT can be as a negative biomarker of outcome of DME.

[Knockout of BMAL1 Gene Induces Apoptosis of HL-60 Cells and Inhibits its Proliferation].

OBJECTIVE: To explore the biological function of BMAL1 in human acute myeloid leukemia by means of the HL-60 cell line in whica circadian gene BMAL1 was konocked-out by the CRISPR/Cas9 technology. METHODS: Two sgRNAs for BMAL1 were designed and the PX459 knockout vectors containing the sgRNA were constructed. The activity of 2 sgRNAs was detected by T7 endonuclease I. the BMAL1 knocked out HL-60 cells were prepared by transient transfection of the target vectors into the cells. Western blot was used to detect the expression of BMAL1 protein. The apoptosis of the targeted cells was detected by flow cytometry. The proliferation status of the cells was assessed by the CCK-8 assay. RESULTS: The PX459-sgRNA vectors were successfully constructed and screened to assure the activity of the targeting vector. It was found that the expression of BMAL1 protein was not detected in BMAL1-knocked out HL- 60 cells. Further, it was shown that BMAL1 knockdout could promote the apoptosis of HL-60 cells and inhibit the cell proliferation ability. CONCLUSION: BMAL1 knocked out HL-60 cells have bean successfully established using the CRISPR/Cas9 gene editing technique, and BMAL1 knockout can promote the HL-60 cell apoptosis and inhibit its proliferation.These result reveal the biological role of the BMAL1 circadian gene in acute myeloid leukemia.

Anti-cancer activity and potential mechanism of a novel aspirin derivative.

Aspirin has been used in the treatment and chemoprevention of many malignant cancers. The mechanism of its anti-cancer activity mainly involves the inhibition of cyclooxygenase-2 (COX-2). However, the application of aspirin is limited by the serious gastric mucosal damage that accompanies its usage. We have previously reported the preparation of a novel aspirin derivative that we named Ca-Asp, and showed that it causes less damage to gastric mucosa of rat and inhibits the expression of COX-2 to higher degree than Asp. However, the anti-cancer effect and mechanism of Ca-Asp was not demonstrated. In this study, the anti-cancer effect of Ca-Asp was investigated and compared with those of Asp and Hydroxyapatite (Hap) at the cell level. The results showed that treatment of SGC-7901 cells (human gastric cancer cell line) with 200-400µg/ml Ca-Asp resulted in significant reduction in cell viability, compared to treatment with either Asp or Hap, and at a higher concentration (500µg/ml). Subsequent investigation into the possible underlying mechanism showed that Ca-Asp induced apoptosis and caused cell cycle arrest at the G1 phase. Ca-Asp also up-regulated the levels of caspase-3 and p53, but down regulated the level of cyclin D1, NF-κB, COX-2 and PGE2. Furthermore, simultaneous treatment of SGC-7901 cells with Ca-Asp and exogenous PGE2 reduced the anti-proliferative effect of Ca-Asp on the cells. Taken together, the results suggested that Ca-Asp might act as a potential anti-cancer drug, and that its suppression of PGE2 production might constitute an important part of its anti-cancer activity.

[Mechanisms of sorafenib induced NB4 cell apoptosis].

OBJECTIVE: To investigate the effects of sorafenib on human acute promyelocytic leukemia cell NB4 and its mechanism. METHODS: The human acute promyelocytic leukemia cell NB4 was treated with different concentrations (0, 1.5, 3, 6 and 12 µmol/L) of sorafenib, the proliferation inhibitory rate of NB4 cells was assayed by MTT, the apoptosis of NB4 was determined with flow-cytomatry after treatment; after extraction of total protein, the Western blot was performed to determine the expressions of apoptosis-relatived molecules Caspase-3, Caspase-8 and MCL-1. The mRNA expressions of Caspase-3, Caspase-8 and MCL-1 were determined by RT-PCR. RESULTS: As compared with the control group, the proliferation of NB4 significantly decreased after treatment with different concentrations of sorafenib. The sorafenib significantly induced the apopotosis of NB4 cells in time- and dose-dependent manners. Furthermore, sorafenib treatment resulted in the obvious increase of the Caspase-3 and Caspase-8 protein and mRNA expressions, and down-regulated the MCL-1 protein and mRNA expressions in NB4 cells. CONCLUSION: Sorafenib can inhibit proliferation and induce apopotosis of human acute promyelocytic leukemia cell NB4 through the expression of Caspase-3 and Caspase-8, and down-regulation of the expression of MCL-1.

[Study on the quality of life and its related factors on patients with multiple myeloma].

OBJECTIVE: To evaluate the quality of life and influencing factors on patients with multiple myeloma (MM). METHODS: 227 MM cases were selected at 5 hospitals in Xi'an from August, 2010 to March, 2013. QLQ-C30 was used to evaluate the quality of life of MM patients, and their norms were as control. Factors which influencing the quality of life were investigated and analyzed with SPSS 17.0 software. RESULTS: The total score of quality of life in MM patients was 49.0±21.7 which was lower than the norms (60.7±23.4). The scores on fatigue, nausea, vomiting, pain, short of breath, disturbance on sleeping, losing appetite, constipation, other symptoms and financial difficulty were significantly higher than data of the norms (P < 0.05). Factors as being elderly (especially those older than 70), under higher proportion of medical costs on their own expense or financial difficulty etc., had major influences on the quality of life (P < 0.05) of MM patients who in particular having worse quality of life when in worsening clinical ISS stage (P < 0.05). Low level of hemoglobin, high level of serum calcium and globulin all significantly reduced the quality of life of the MM patients (P < 0.05). CONCLUSION: The quality of life of MM patients was significantly lower than the normal people or patients with other tumors. Fatigue, pain, and financial difficulty were main influencing factors on the quality of life of MM patients. The assessment on the effects of treatment should relate to the improvement of hemoglobin, serum calcium and globulin, which could all improve the quality of life of MM patients.

[Caspase-independent apoptosis induced by arsenic trioxide in human multiple myeloma cell RPMI8226].

This study was purposed to explore the caspase-independent apoptosis pathway in human multiple myeloma cell RPMI8226 induced by arsenic trioxide (As(2)O(3)). MTT method was used to analyze the proliferation inhibition rate; flow cytometry was used to detect the apoptosis rate; Western blot was used to determine the expressions of BCL-2 and Caspase-3 in RPMI8226 cells. The results showed that As(2)O(3) (0.1 - 20 µmol/L) significantly inhibited the proliferation of RPMI8226 (P < 0.05) in concentration- and time-dependent manner. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, the apoptosis rate of zVAD-fmk (20 µmol/L) and As(2)O(3) combined treated group did not change. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, zVAD-fmk (20 µmol/L) combined with As(2)O(3) (10 µmol/L) treatment group showed significant increase of expressions of Caspase-3 and BCL-2. It is concluded that As(2)O(3) can inhibit the proliferation of RPMI8226 cells. As(2)O(3) can induce apoptosis of RPMI8226 cells, and a caspase-independent process probably exist in As2O3-inducing RPMI8266 cells apoptosis.

[The efficacy and safety of HAA regimen as induction chemotherapy in 150 newly diagnosed acute myeloid leukemia].

OBJECTIVE: To explore the efficacy and safety of HAA regimen (homoharringtonine, cytarabine and aclarubicin) in the treatment of 150 newly diagnosed adult acute myeloid leukemia (AML). METHODS: All patients entered the study from May 1999 to June 2008 were treated with HAA regimen. Cox-survival analysis was used to estimate the survival rate and differences between M(1)/M(2) and M(4)/M(5) were compared with 2-sided log-rank test. RESULTS: Out of the 150 patients, 121 (81%) achieved complete remission (CR). After the first course, CR rate was 68%. The CR rates of 97%, 84% and 38% were achieved in patients with favorable, intermediate and unfavorable cytogenetics, respectively. For the patients with CR, the median follow-up time was 16.5 (1.5 - 100.5) months, and the estimated 3-year survival rate was 45%. The estimated 3-year relapse free survival rate was 52% for the 121 patients with CR. CONCLUSIONS: HAA regimen may be an efficacious and safe regimen with a good toleration in the induction therapy for newly diagnosed AML, and a high CR rate could be achieved with only one or two courses.

[Effect of recombinant human erythropoietin on bcl-2 protein expression in the retina in a rabbit model of acute high intraocular pressure].

OBJECTIVE: To investigate the effect of recombinant human erythropoietin (rhEPO) on the expression of bcl-2 protein in the retina of rabbits with acute high intraocular pressure and explore the mechanism underlying the protective effect of rhEPO on the retina against ischemia-reperfusion injury. METHODS: rhEPO was injected subcutaneously in the ear of a rabbit model of acute high intraocular pressure induced by physiological saline perfusion into the anterior chamber. Bcl-2 protein expression in the retina of the rabbits was observed by immunohistochemical staining on days 1, 3, 7, and 14 after retinal ischemia-reperfusion and compared with that in normal rabbits and untreated rabbit models. RESULTS: bcl-2-positive cells were observed in the retina of normal rabbits with a mean positive cell number of 10.5-/+1.2 in each high-power visual field. Compared with that in the normal control group, the number of the positive cells decreased significantly in both the model group and EPO group (P<0.05, P<0.01), but the latter group showed a significantly greater number than the former (P<0.05 at day 7 and P<0.01 at day 14). CONCLUSION: Systemic administration of rhEPO can up-regulate the expression of bcl-2 protein in the retina of rabbits with acute high intraocular pressure, which is probably one of the mechanisms for the protective effect of rhEPO on the retina against ischemia-reperfusion injury.