[Copy number variations of CCND1 gene and chromosome 11 centromere in acral melanoma].

Objective: To study the correlation between the copy number variations of CCND1 gene and chromosome 11 and their associations with clinicopathologic features in acral melanoma. Methods: Thirty-three acral melanoma cases diagnosed at the Department of Pathology of Peking University Third Hospital, Beijing, China from January 2018 to August 2021 were collected. Fluorescence in situ hybridization (FISH) was used to detect the copy number of CCND1 gene and centromere of chromosome 11. The relationship between the copy numbers of CCND1 and chromosome 11 centromere, and the correlation between CCND1 copy number and clinicopathologic characteristics were analyzed. Results: There were 15 male and 18 female patients, with an age ranging from 22-86 years. 63.6% (21/33) of the patients had an increased CCND1 gene copy number. 21.2% (7/33) of patients with increased CCND1 copy number had an accompanying chromosome 11 centromere copy number increase. 27.3% (9/33) of the cases had a low copy number of CCND1 gene, and 4 of them (4/33, 12.1%) were accompanied by chromosome 11 centromere copy number increase. 36.4% (12/33) of the cases had a high copy number of CCND1 gene, and 3 (3/33, 9.1%) of them were accompanied by chromosome 11 centromere copy number increase. No cases with CCND1 low copy number increase showed CCND1/CEP11 ratio greater than 2.00. The 11 cases with CCND1 high copy number increase showed CCND1/CEP11 ratio greater than or equal to 2.00. However, there was no significant correlation between CCND1 copy number increase and any of the examined clinicopathologic features such as age, sex, histological type, Breslow thickness, ulcer and Clark level. Conclusions: CCND1 copy number increase is a significant molecular alteration in acral melanoma. In some cases, CCND1 copy number increase may be accompanied by the copy number increase of chromosome 11. For these cases the copy number increase in CCND1 gene may be a result of the copy number change of chromosome 11.

[miR-148b inhibits M2 polarization of LPS-stimulated macrophages by targeting DcR3].

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.

[In vitro pharmacodynamic studies of novel class â and â ¡b selective histone deacetylase inhibitor purinostat mesylate in the treatment of diffuse large B-cell lymphoma and its mechanism].

Objective: To investigate the in vitro inhibitory activity of a novel class Ⅰ and Ⅱb selective histone deacetylase (HDAC) inhibitor, purinostat mesylate (PM) , in diffuse large B-cell lymphoma and its mechanism. Methods: The 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide method was used to detect the effect of PM on cell proliferation. The effects of PM on cell cycle and apoptosis were detected by flow cytometry. The acetylation levels of HDAC substrate, cell cycle protein, apoptosis-related protein, and oncogene protein expression were detected by Western blot. Results: PM significantly inhibited the proliferation of lymphoma SUDHL-4 and SUDHL-6 cells and increased the acetylation levels of HDAC substrates H3, H4, and α-tubulin. In cell cycle experiments, PM induced G(0)/G(1) phase arrest in SUDHL-4 and SUDHL-6 cells. Western blot experiment showed that PM could significantly downregulate the expression of cyclin-dependent kinases Cdk2, Cdk4, Cdk6, cyclin D1, and cyclin E and upregulate the expression of CDK inhibitor protein p21. In the apoptosis experiment, PM could induce the apoptosis of SUDHL-4 and SUDHL-6 cells. Western blot experiment demonstrated that PM promoted endogenous apoptosis by activating caspase-3 kinase and affecting antiapoptotic protein Bcl-2. In addition, PM could downregulate the expression of oncogene marker proteins MYC, IKZF1, and IKZF3. Conclusion: PM has an efficient biological activity in vitro for diffuse large B-cell lymphoma, including double-hit lymphoma, and provides valuable experimental evidence for PM in clinical treatment.

[Establishment of an early risk prediction model for bloodstream infection and analysis of its predictive value in patients with extremely severe burns].

Objective: To establish an early prediction model for bloodstream infection in patients with extremely severe burns based on the screened independent risk factors of the infection, and to analyze its predictive value. Methods: A retrospective case-control study was conducted. From January 1, 2010 to December 31, 2019, 307 patients with extremely severe burns were admitted to the Department of Burns and Plastic Surgery of Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medcine, including 251 males and 56 females, aged from 33 to 55 years. According to the occurrence of bloodstream infection, the patients were divided into non-bloodstream infection group (221 cases) and bloodstream infection group (86 cases). The gender, age, body mass index, outcome, length of hospital stay of patients were compared between the two groups, and the detection of bacteria in blood microbial culture of patients was analyzed in bloodstream infection group. The included 307 patients were divided into modeling group (219 cases) and validation group (88 cases) according to the random number table with a ratio of about 7∶3. The gender, age, body mass index, total burn area, full-thickness burn area, combination of inhalation injury, implementation of mechanical ventilation, days of mechanical ventilation, days of intensive care unit (ICU) stay, outcome, length of hospital stay, complication of bloodstream infection of patients were compared between the two groups. According to the occurrence of bloodstream infection, the patients in modeling group were divided into bloodstream infection subgroup (154 cases) and non-bloodstream infection subgroup (165 cases). The total burn area, full-thickness burn area, combination of inhalation injury, implementation of mechanical ventilation, days of mechanical ventilation, and days of ICU stay of patients were compared between the two subgroups. The above-mentioned data between two groups were statistically analyzed with one-way analysis of independent sample t test, chi-square test, and Mann-Whitney U test to screen out the factors with statistically significant differences in the subgroup univariate analysis of modeling group. The factors were used as variables, and binary multivariate logistic regression analysis was performed to screen out the independent risk factors of bloodstream infection in patients with extremely severe burns, based on which the prediction model for bloodstream infection in patients with extremely severe burns of modeling group was established. The receiver operating characteristic (ROC) curve of the prediction model predicting the risk of bloodstream infection of patients in modeling group was drawn, and the area under the ROC curve was calculated. The sensitivity, specificity, and the best prediction probability were calculated according to the Youden index. According to the occurrence of bloodstream infection, the patients in validation group were divided into bloodstream infection subgroup (21 cases) and non-bloodstream infection subgroup (67 cases). The prediction probability >the best prediction probability of model was used as the judgment standard of bloodstream infection. The prediction model was used to predict the occurrence of bloodstream infection of patients in the two subgroups of validation group, and the incidence, specificity, and sensitivity for predicting bloodstream infection were calculated. In addition, the ROC curve of the prediction model predicting the risk of bloodstream infection of patients in validation group was drawn, and the area under the ROC curve was calculated. Results: Compared with those of non-bloodstream infection group, the mortality of patients in bloodstream infection group was significantly higher (χ2=8.485, P<0.01), the length of hospital stay was significantly increased (Z=-3.003, P<0.01), but there was no significant change in gender, age, or body mass index (P>0.05). In patients of bloodstream infection group, 110 strains of bacteria were detected in blood microbial culture, among which Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii were the top three bacteria, accounting for 35.45% (39/110), 26.36% (29/110), and 13.64% (15/110), respectively. Gender, age, body mass index, total burn area, full-thickness burn area, proportion of combination of inhalation injury, proportion of implementation of mechanical ventilation, days of mechanical ventilation, days of ICU stay, outcome, length of hospital stay, and proportion of complication of bloodstream infection of patients were similar between modeling group and validation group (P>0.05). Compared with those of non-bloodstream infection subgroup in modeling group, the total burn area, full-thickness burn area, proportion of combination of inhalation injury, proportion of implementation of mechanical ventilation, days of mechanical ventilation, and days of ICU stay of patients in bloodstream infection subgroup were significantly increased (Z=-4.429, t=-4.045, χ2=7.845, 8.845, Z=-3.904, -4.134, P<0.01). Binary multivariate logistic regression analysis showed that total burn area, days of ICU stay, and combination of inhalation injury were the independent risk factors for bloodstream infection of patients in modeling group (odds ratio=1.031, 1.018, 2.871, 95% confidence interval=1.004-1.059, 1.006-1.030, 1.345-6.128, P<0.05 or P<0.01). In modeling group, the area under the ROC curve was 0.773 (95% confidence interval=0.708-0.838); the sensitivity was 64.6%, the specificity was 77.9%, and the best prediction probability was 0.335 when the Youden index was 0.425. The bloodstream infection incidence of patients predicted by the prediction model in validation group was 27.27% (24/88), with specificity of 82.09% (55/67) and sensitivity of 57.14% (12/21). The area under the ROC curve in validation group was 0.759 (95% confidence interval=0.637-0.882). Conclusions: The total burn area, days of ICU stay, and combination of inhalation injury are the risk factors of bloodstream infection in patients with extremely severe burns. The early prediction model for bloodstream infection risk in patients with extremely severe burns based on these factors has certain predictive value for burn centers with relatively stable treatment methods and bacterial epidemiology.

[The expression of Notch pathway in nasal polyps and its correlation with the decreased Treg percentage and eosinophil infiltration].

Objective: To investigate the correlation between Notch pathway expression in nasal polyps and Treg percentage and Eos infiltration. Methods: Patients with chronic sinusitis and simple nasal septum deviation who received nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-Sen University between November 2012 and August 2018 were selected and enrolled in CRS group and control group respectively. Nasal mucosa tissues were collected from 30 CRSsNP patients (14 males and 16 females aged from 18 to 63), 58 CRSwNP patients (38 males and 20 females aged from 18 to 65) and 29 patients (19 males and 10 females aged from 20 to 57), who underwent nasal endoscopic surgery for correction of simple nasal septum deviation. Hematoxylin-eosin(HE) staining was used to observe the infiltration of eosinophilic granulocytes in the tissues and to classify chronic sinusitis with polyps (CRSwNP) into eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP)and non-eosinophilic chronic rhinosinusitis with nasal polyps (Eos-CRSwNP). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Notch pathway receptors (Notch-l, 2, 3, 4) and their ligands (Jagded-l, Jagded-2, Delta-l, Delta-3and Delta-4) in the nasal mucosa of each group, as well as the expression of Th2 cytokines (IL-4, IL-5, IL-13), eosinophilic cationic protein (ECP)and the key transcription factor Foxp3 in Treg cells. Finally, flow cytometry was used to detect CD4+CD25+Foxp3+ Treg cells in nasal mucosa of each group. Results: Compared with controls, the expression of Th2 cytokines (IL-4, IL-5, IL-13) in CRSsNP and non-Eos-CRSwNP patients was the highest in Eos-CRSwNP (F=16.930,9.197,9.116, all P<0.05). Foxp3 had the lowest expression in Eos-CRSwNP patients and was lower than non-Eos-CRSwNP patients (F=2.780,P<0.05), and was negatively correlated with ECP (r=-0.326,P<0.05). Compared with controls, Eos-CRSwNP patients in CRSsNP patients and non-Eos-CRSwNP patients exhibited a significantly lower frequency of CD4+CD25+Foxp3+Treg cells (F=13.140, all P<0.01). The expression of Notch-l and Jagged-l in Eos-CRSwNP was significantly higher than that of the controls, CRSsNP patients and non-Eos-CRSwNP patients (F=5.953/F=6.380, P<0.05). In the nasal polyp group, the expression of Notch-l and Jagged-l showed significantly negative correlation with Foxp3 (r=-0.611/-0.346, all P<0.05), and positive correlation with Th2 cytokines (IL-4, IL-5, IL-13) and ECP, respectively (r=0.781/0.459,0.621/0.601,0.605/0.490,0.464/0.668, all P<0.05). There was no significant difference in the expression of receptor and ligand of the other Notch pathway among the groups. Conclusion: Abnormal activation of Notch-l/Jagged-l pathway may be involved in decreasing Treg ratio in Eos-CRSwNP, thereby promoting Th2 inflammatory response and Eosinophil infiltration.

[The advances in surgery for cirrhotic portal hypertension in China].

The cirrhotic portal hypertension is very common worldwide and poses a serious threat to the health of patients.Over past three decades, the surgical treatment for cirrhotic portal hypertension was strongly challenged by the drugs, endoscopy, interventional therapy and liver transplantation.However, under the multidisciplinary team(MDT) cooperative diagnosis and treatment mode, the surgical treatment still plays a unique and irreplaceable role.Laparoscopic pericardial vascular devascularization is characterized by less injury and bleeding, rapid postoperative recovery, which will coexist with open surgery for portal hypertension. It is important to focus on the development and application of new methods, new technologies and new concepts under the MDT cooperative diagnosis and treatment mode, giving full play to the advantages of each discipline and advocate standardized, individualized and precise treatment should be emphasized to maximize patient clinical benefits.

[The progresses of liver resection for solitary large hepatocellular carcinoma].

Large hepatocellular carcinoma (HCC) is one of the most common malignancies and was mistaked as "advanced and unresectable" . Liver resection is still the best curable treatment for HCC.The resection of large HCC is very difficult, which seriously restrict the progress of liver surgery.Our study proved that solitary large HCC (SLHCC) has unique clinicopathological and molecular biological characteristics.No matter how big the tumor size is, it belongs to early stage if there is no vascular invasion.Liver resection should be aggressively recommended for the patients with SLHCC, in which they can obtain good outcome, with 40% 5-year survival rate.We has also defined the borderline resectable hepatocellular carcinoma, and suggested that strictly master and correctly judge the surgical indications, syntheticly evaluate the surgical safety and patient's tolerability for liver resection.After that, with hands of experienced surgeons, liver resection for SLHCC can be safely and reliablely performed.

Synovial fluid interleukin-16, interleukin-18, and CRELD2 as novel biomarkers of prosthetic joint infections.

OBJECTIVES: Prosthetic joint infection (PJI) diagnosis is a major challenge in orthopaedics, and no reliable parameters have been established for accurate, preoperative predictions in the differential diagnosis of aseptic loosening or PJI. This study surveyed factors in synovial fluid (SF) for improving PJI diagnosis. METHODS: We enrolled 48 patients (including 39 PJI and nine aseptic loosening cases) who required knee/hip revision surgery between January 2016 and December 2017. The PJI diagnosis was established according to the Musculoskeletal Infection Society (MSIS) criteria. SF was used to survey factors by protein array and enzyme-linked immunosorbent assay to compare protein expression patterns in SF among three groups (aseptic loosening and first- and second-stage surgery). We compared routine clinical test data, such as C-reactive protein level and leucocyte number, with potential biomarker data to assess the diagnostic ability for PJI within the same patient groups. RESULTS: Cut-off values of 1473 pg/ml, 359 pg/ml, and 8.45 pg/ml were established for interleukin (IL)-16, IL-18, and cysteine-rich with EGF-like domains 2 (CRELD2), respectively. Receiver operating characteristic curve analysis showed that these factors exhibited an accuracy of 1 as predictors of PJI. These factors represent potential biomarkers for decisions associated with prosthesis reimplantation based on their ability to return to baseline values following the completion of debridement. CONCLUSION: IL-16, IL-18, and CRELD2 were found to be potential biomarkers for PJI diagnosis, with SF tests outperforming blood tests in accuracy. These factors could be useful for assessing successful debridement based on their ability to return to baseline values following the completion of debridement.Cite this article: M-F. Chen, C-H. Chang, L-Y. Yang, P-H. Hsieh, H-N. Shih, S. W. N. Ueng, Y. Chang. Synovial fluid interleukin-16, interleukin-18, and CRELD2 as novel biomarkers of prosthetic joint infections. Bone Joint Res 2019;8:179-188. DOI: 10.1302/2046-3758.84.BJR-2018-0291.R1.

[Prevalence of dyslipidemia and risk factors among coal miners under different work conditions].

Objective: To understand the prevalence of dyslipidemia and risk factors among coal miners under different work conditions. Methods: The survey was conducted from April 2016 to June 2016. 759 mine workers were divided into three groups (group of the front line miner, underground auxiliary and ground) . Questionnaire and physical examination were used to collect related information of workers. Logistic regression model was used to analyze relative factors. Results: The overall prevalence of dyslipidemia was 43.2% in coal miners. The prevalence rate of the front line miner and underground auxiliary miners was 46.6%. Ground workers had the lowest prevalence rate of 36.4%. Multiple Logistic regression analysis showed that higher body mass index (BMI) was risk factors for underground workers (OR=2.18, 95%CI:1.51~3.13) . Smoking (OR=1.99, 95%CI:1.17~3.38) , drinking (OR=1.85, 95%CI:1.11~3.06) , hypertension (OR=1.79, 95%CI:1.00~3.22) and higher waist and hip ratio (OR=1.06, 95%CI:1.04~1.09) were risk factors for underground auxiliary workers. For ground workers, those with higher BMI (OR=2.64, 95%CI:1.68~4.16) were at higher risk of dyslipidemia and female workers had lower risk (OR=0.35, 95%CI:0.18~0.65) than male workers. Conclusion: The dyslipidemia rate of coal mine workers is related to work environment and behavior. Health education may be needed to reduce the dyslipidemia rate of coal mine workers.

Patient-reported outcome use in oncology: a systematic review of the impact on patient-clinician communication.

PURPOSE: Patient-reported outcomes (PROs) are an increasingly popular tool to optimize care and bridge the gap between patient experience and clinician understanding. The aim of this review was to identify mechanisms through which PROs facilitate patient-clinician communication in the adult oncology population. METHODS: We conducted a systematic review of the published literature using the following data sources: MEDLINE, EMBASE, CINAHL, PsycINFO, Cab Direct, and CDSR. Studies included in this review reported on the outcomes of PRO use, used PROs as an intervention and not as a study outcome measurement tool, included cancer patients or survivors as study participants, and analyzed patient-clinician communication. RESULTS: We identified 610 unique records, of which 43 publications met the inclusion and exclusion criteria. Synthesis of the reviewed studies provided evidence of the usefulness of PROs in facilitating patient-clinician communication on a variety of topics. We identified mechanisms though which PROs influenced patient-clinician communication to include increasing symptom awareness, prompting discussion, streamlining consultations, and facilitating inter-professional communication. Barriers to PRO use in communication improvement include technical problems impeding its administration and completion, compliance issues due to lack of incentive or forgetfulness, and use of PROs that do not appropriately assess issues relevant to the patient. Facilitators include increased education on PRO use, using PRO tools that patients find more acceptable, and providing patient data summaries in an easily accessible format for clinicians. CONCLUSIONS: Our review suggests that PROs facilitate patient-clinician communication through various mechanisms that could perhaps contribute to improvements in symptom management and survival. The impact of PROs on clinical outcomes, however, remains poorly studied.

Neuropilin-1 contributes to esophageal squamous cancer progression via promoting P65-dependent cell proliferation.

Neuropilin-1 (NRP1) is a non-kinase receptor recently implicated in tumor progression. Here we revealed that over-expression of NRP1 correlates with poor prognosis in esophageal squamous cell carcinoma (ESCC). NRP1-knockdown suppressed ESCC cell proliferation and xenograft tumor growth. Reduced NRP1 expression downregulated P65 mRNA and protein expression, and ectopic expression of P65-restored cell proliferation in NRP1-silenced cells. NRP1 regulates P65 transcription by activating cAMP responsive element binding protein (CREB). NRP1 interacted with and activated epidermal growth factor receptor (EGFR), and b1/b2 domain of NRP1 is responsible for the activation of EGFR. We also found that EGFR regulated CREB transcriptional activity via AKT. These data suggest that NRP1 is an upstream regulator in the P65-dependent proliferation signaling pathway and a candidate therapeutic target for ESCC.

[Study on autophagy in nucleated red blood cells in patients with myelodysplastic syndromes].

Objective: To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Methods: Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA(+) nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA(+) nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA(+) nucleated RBC was detected by Western blot. Results: Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA(+) nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA(+) nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA(+) nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Conclusion: Autophagy is impaired in nucleated RBC of MDS patients.

[NIX-mediated mitochondrial autophagy in pathogenesis of myelodysplastic syndrome anemia].

Objective: To investigate the change of NIX level of bone marrow nucleated red blood cells in anemia patients with myelodysplastic syndromes (MDS), to explore the significance of NIX-mediated mitochondrial autophagy in the pathogenesis of MDS anemia. Methods: A total of 54 patients with MDS diagnosed in the Department of Hematology of General Hospital, Tianjin Medical University from July 2015 to July 2016 were enrolled into the MDS group, 33 cases of immune thrombocytopenia or idiopathic leukopenia as controls.The level of NIX, the number of mitochondria, mitochondrial membrane potential, the level of reactive oxygen species (ROS) in GlycoA(+) nucleated red blood cells were measured by flow cytometry; the level of NIX mRNA was measured by PCR. Results: (1) The expression of NIX in GlycoA(+) nucleated red blood cells in high-risk MDS patients (0.61±0.24) was significantly lower than that in controls (0.79±0.16, P=0.027), and lower than that in low-risk MDS patients (0.81±0.15, P=0.011), while there was no significant difference between the controls and low-risk MDS patients. The expression of NIX mRNA in GlycoA(+) nucleated red blood cells in high-risk MDS group (0.36±0.09) was lower than that in the controls (1.44±0.41, P=0.027) and that in the low-risk group (1.02±0.22, P=0.012); there was no significant difference between the controls and the low-risk group. (2) The number of mitochondria in GlycoA(+) nucleated red blood cells in high-risk MDS patients (937.17±707.85) was significantly higher than that in the controls (513.49±372.33, P=0.019) and that in low-risk MDS patients (461.74±438.02, P=0.008); while there was no significant difference between low-risk MDS patients and the controls. (3) The level of mitochondrial membrane potential in GlycoA(+) nucleated red blood cells in high-risk MDS patients (0.33±0.18) was significantly lower than that in the controls (0.61±0.32, P=0.001) and that in low-risk MDS patients (0.61±0.34, P=0.001); with no significant difference between low-risk MDS patients and the controls. (4)The level of ROS in GlycoA(+) nucleated red blood cells in high-risk MDS patients (438.65±322.83) was significantly higher than that in the controls (242.77±136.87, P=0.006), and higher than that in low-risk MDS patients (197.40±95.07, P=0.001); no significantly different between low-risk MDS patients and the controls. (5) The number of mitochondria in GlycoA(+) nucleated red blood cell was positively correlated with the percentage of ring sideroblast (r=0.457, P=0.028) in the MDS patients.(6) The number of mitochondria in GlycoA(+) nucleated red blood cells was negatively correlated with the concentration of hemoglobin (r=-0.521, P=0.009) in high-risk MDS patients, but not correlated with the concentration of hemoglobin in low-risk MDS patients. Conclusion: NIX level is reduced in nucleated red blood cells of high-risk MDS patients, which leads to impaired mitochondrial autophagy, increased damaged mitochondria and apoptosis of nucleated red blood cells, thus related with anemia.

[Abnormality of TOP2A expression and its gene copy number variations in neuroblastic tumors].

Objective: To detect TOP2A protein expression and gene copy number alterations, and to analyze related clinical and pathological implications in pediatric neuroblastic tumors (NT). Methods: Immunohistochemistry was used to detect TOP2A protein expression. Fluorescence in situ hybridization (FISH) was used to detect numerical aberrations of TOP2A. Results: TOP2A protein was expressed in 59.1%(52/88) of cases, which was associated with differentiation (P=0.006), Ki-67 index (P<0.01) and MKI (P=0.001). Twenty-eight cases (35.0%, 28/88) showed TOP2A gene amplification, which was correlated with the age (P<0.01), clinical stage (P=0.028), high risk group (P=0.001), Ki-67 index (P=0.040) and differentiation (P=0.014). Survival analysis showed that TOP2A expression was related to survival rate. Multivariate analyses showed that TOP2A expression was an independent predictor for poor prognosis (P=0.010). Conclusions: More than half of the cases show TOP2A expression, which is more likely associated with NB, high Ki-67 index and high MKI. Cases with TOP2A expression have shorter survivals and poorer prognosis. TOP2A amplification is seen in 35% and likely occurs in patients older than 18 months and at advanced INSS stages (Ⅲ and Ⅳ). As a target of the anthracycline-based adjuvant drugs, TOP2A test can be used to select patient with NT for the therapy.

[Pulmonary surfactant protein adenosine triphosphate-binding-cassette-A3 gene composite mutations in infant congenital interstitial lung disease: report of a case and review of literature].

Objective: To report a case of the pulmonary surfactant protein(SP) adenosine triphosphate-binding-cassette-A3 (ABCA3) gene mutations in infant congenital interstitial lung disease(ILD), and review the related literature, to investigate the relationships of ABCA3 gene mutation associated with ILD in infants. Method: A 6-months-old boy was hospitalized in the department of Pediatrics of the First Affiliated Hospital of Guangzhou Medical University. The clinical, radiological, histological information from transbronchial lung biopsy (TBLB) and genetic testing in this case was analyzed; 12 reports retrieved on literature search at Pubmed, OVID databases from 2004 to 2015 by using the ABCA3 as keyword were reviewed and analyzed. Result: (1)The patient, a 6-months-old boy, had progressive tachypnea and dyspnea since 4 months old. Physical examination on admission revealed respiratory rate of 78 times/min , heart rate of 187 times/min, SpO2 0.93(mask oxygen-inspiration with 6 L/min), scattered fine moist crackles could be heard over the both lungs, clubbing fingers were found. High-resolution computed tomography(HRCT) revealed diffuse ground-glass opacity, interlobular and intralobular septal thickening. Lung biopsies showed evidences of the alveolar cavity atelectatic changes and interstitial fibrosis. SP-A and SP-B were negative in immunohistochemical stainting. SP-related gene sequence analysis found that there was compound heterozygous missense mutation of ABCA3 gene in c. 1942A>G, c.2701-33G>C and c. 991-105C>A. (2)The review of related literature found that totally 12 cases were reported. The main manifestations were progressive tachypnea and dyspnea, age of onset was between birth and 4 years of age. The imaging characteristics of chest HRCT revealed diffuse infiltration or diffuse ground-glass pattern in the lung. PROGNOSIS: 6 cases died, and 6 cases survived, including 4 cases with pulmonary function disturbance to different degrees; 12 cases had ABCA3 gene mutations, 9 cases had composite ABCA3 gene mutations, in 11 cases the mutation occured in the exon of coding region, in 1 case in the intron, 9 cases had heterozygous mutations, 3 cases had homozygous mutations. Conclusion: The main phenotypes of ABCA3 mutation associated with ILD were full term neonatal respiratory distress syndrome or progressive tachypnea or dyspnea unexplained in infants. The chest HRCT showed two diffuse pulmonary interstitial changes. ABCA3 mutation mainly was multi-site composite mutations and heterozygous mutations in the exon of coding region.

[Prognostic factors analysis for the long-term survival in solitary large hepatocellular carcinoma patients after hepatectomy].

OBJECTIVE: To explore the prognostic factors that have affected the long-term survival of solitary large hepatocellular carcinoma (SLHCC) patients after hepatectomy. METHODS: The clinical data of 215 SLHCC patients accepted hepatectomy in the Xiangya Hospital, Central South University from January 2004 to December 2012 were retrospectively analyzed. There were 182 males and 33 females aged from 24 to 69 years(median age was 46 years). Using a variety of statistical methods, including the Kaplan-Meier estimator and the Log-rank test, the impacts of an array of clinicopathologic factors, such as age, gender, liver cirrhosis, chronic viral hepatitis, the Child-Pugh grading, microvascular invasion, macrovascular invasion and TNM staging, on the overall survival and the disease-free survival of SLHCC patients after hepatectomy were analyzed.The prognostic factors were evaluated by univariate and multivariate analyses for the long-term survival of SLHCC patients after hepatectomy. RESULTS: The whole group of patients with SLHCC showed 1-, 3-, and 5-year overall survival rates of 88.1%, 60.2%, and 41.7%, respectively, and exhibited 1-, 3-, and 5-year disease-free survival rates of 80.1%, 49.4%, and 33.6%, respectively. The 1-, 3-, and 5-year overall survival rates and disease-free survival rates of SLHCC patients with microvascular invasion were 82.0%, 45.1%, 29.0% and 69.6%, 36.1%, 23.5%, respectively. In addition, the 1-, 3-, and 5-year overall survival rates and disease-free survival rates of SLHCC patients with macrovascular invasion were 64.7%, 34.3%, 18.3% and 54.2%, 24.1%, 0, respectively. In contrast, the 1-, 3-, and 5-year overall survival rates and disease-free survival rates of SLHCC patients without vascular invasion were 95.0%, 72.3%, 51.8% and 90.1%, 60.9%, 42.9%, respectively. The results of univariate analysis indicated that liver cirrhosis, microvascular invasion, macrovascular invasion, TNM staging were hazardous factors for the overall survival of SLHCC patients(χ(2)=4.953, 8.835, 15.237, 19.789 respectively, all P<0.05); while microvascular invasion, macrovascular invasion, TNM staging were risk factors for the disease-free survival of SLHCC patients(χ(2)=12.974, 13.247, 24.516 respectively, all P<0.05). Furthermore, the multivariate analysis suggested that microvascular invasion, macrovascular invasion, TNM staging were the independent prognostic factors that have affected the overall survival and disease-free survival of SLHCC patients(all P<0.05). CONCLUSION: Microvascular invasion, macrovascular invasion and TNM staging were the independent prognostic factors for the long-term survival of patients with SLHCC after hepatectomy.

Influence of menopausal status on diagnostic accuracy of myometrial invasion in endometrial cancer: diffusion-weighted and dynamic contrast-enhanced MRI at 3 T.

AIM: To assess the influence of menopausal status on the diagnostic accuracy of diffusion-weighted (DW) and dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) for evaluating myometrial invasion in patients diagnosed with endometrial cancer. MATERIALS AND METHODS: In this prospective study, 91 consecutive female patients diagnosed with endometrial cancer were enrolled for preoperative evaluation using 3 T MRI. Two radiologists interpreted myometrial invasion depth on DW (b=1000 s/mm(2)) and DCE MRI images, with surgical histopathology as the reference standard. Statistical methods included kappa statistics for evaluating reader agreement and diagnostic performance analysis between pre- and postmenopausal groups. RESULTS: Reader agreement of DW MRI was poor (κ=0.20) for premenopausal patients. The diagnostic accuracy of DW MRI in detecting myometrial involvement was significantly lower in premenopausal compared with postmenopausal patients (0.42 versus 0.73, p=0.006). There was no difference in the diagnostic accuracy of DW MRI in detecting deep myometrial invasion between premenopausal and postmenopausal groups (0.94 versus 0.95, p>0.99). CONCLUSION: For premenopausal patients who plan to receive fertility-preserving treatment for endometrial cancer, DCE MRI is superior to DW MRI in excluding any possible myometrial invasion. For preoperative assessment of deep myometrial invasion, DW MRI can be a legitimate alternative to DCE MRI regardless of menopausal status and is particularly beneficial for patients at risk of nephrogenic systemic fibrosis.

Changes of masseter muscle activity following injection of botulinum toxin type A in adult rats.

OBJECTIVES: To investigate changes in masseter muscle function following intramuscular injection of different dose-dependent botulinum toxin type A (BTXA). SETTING AND SAMPLE POPULATION: Department of Orthodontics at Taipei Medical University. Fifty-two, 70-day-old male Wistar rats were randomly divided into four groups. Group I received 7.5 U of BTXA (0.3 ml), Group II received 5.0 U, and Group III received 2.5 U in the right masseter muscle, respectively. Group IV is the control and received no BTXA injection. MATERIALS AND METHODS: A wire electrode device was implanted to record muscle activity. One week after implantation, the rats were fed every 2 h and EMG signals were recorded during the first hour. All signals were recorded for 12 weeks. Thereafter, EMG data were analyzed for statistical calculation and weights of masseter muscles were measured. RESULTS: Masseter muscle activity decreased 99% during the first week after BTXA injection and gradually recovered from the 3rd week on in Groups I-III. By the 12th week, muscle activity recovered to 41% in Groups I and II and 56.26% in Group III. No significant changes of muscle activity were observed in Group IV. CONCLUSION: BTXA induced a reduction in masseter muscle activity and an increased toxin dose resulted in greater depression of muscle activity.