Head and neck MRI-based T stage and [18F]FDG PET/CT-based N/M stage improved prognostic stratification in primary nasopharyngeal carcinoma.

OBJECTIVES: To evaluate whether MRI-based T stage (TMRI), [18F]FDG PET/CT-based N (NPET/CT), and M stage (MPET/CT) are superior in NPC patients' prognostic stratification based on long-term survival evidences, and whether TNM staging method involving TMRI + NPET/CT + MPET/CT could improve NPC patients' prognostic stratification. METHODS: From April 2007 to December 2013, 1013 consecutive untreated NPC patients with complete imaging data were enrolled. All patients' initial stages were repeated based on (1) the NCCN guideline recommended "TMRI + NMRI + MPET/CT" ("MMP") staging method; (2) the traditional "TMRI + NMRI + Mconventional work-up (CWU)" ("MMC") staging method; (3) the single-step "TPET/CT + NPET/CT + MPET/CT" ("PPP") staging method; or (4) the "TMRI + NPET/CT + MPET/CT" ("MPP") staging method recommended in present research. Survival curve, ROC curve, and net reclassification improvement (NRI) analysis were used to evaluate the prognosis predicting ability of different staging methods. RESULTS: [18F]FDG PET/CT performed worse on T stage (NRI = - 0.174, p < 0.001) but better on N (NRI = 0.135, p = 0.004) and M stage (NRI = 0.126, p = 0.001). The patients whose N stage upgraded by [18F]FDG PET/CT had worse survival (p = 0.011). The "TMRI + NPET/CT + MPET/CT" ("MPP") method performed better on survival prediction when compared with "MMP" (NRI = 0.079, p = 0.007), "MMC" (NRI = 0.190, p < 0.001), or "PPP" method (NRI = 0.107, p < 0.001). The "TMRI + NPET/CT + MPET/CT" ("MPP") method could reclassify patients' TNM stage to a more appropriate stage. The improvement is significant in patients with more than 2.5-years follow-up according to the time-dependent NRI values. CONCLUSIONS: The MRI is superior to [18F]FDG PET/CT in T stage, and [18F]FDG PET/CT is superior to CWU in N/M stage. The "TMRI + NPET/CT + MPET/CT" ("MPP") staging method could significantly improve NPC patients' long-term prognostic stratification. CLINICAL RELEVANCE STATEMENT: The present research provided long-term follow-up evidence for benefits of MRI and [18F]FDG PET/CT in TNM staging for nasopharyngeal carcinoma, and proposes a new imaging procedure for TNM staging incorporating MRI-based T stage and [18F]FDG PET/CT-based N and M stage, which significantly improves long-term prognostic stratification for patients with NPC. KEY POINTS: • The long-term follow-up evidence of a large-scale cohort was provided to evaluate the advantages of MRI, [18F]FDG PET/CT, and CWU in the TNM staging of nasopharyngeal carcinoma. • A new imaging procedure for TNM stage of nasopharyngeal carcinoma was proposed.

WeChat-based remote follow-up management alleviates the home care burden and anxiety of parents of premature infants: Randomized controlled study.

OBJECTIVE: The purpose of this study was to explore the effect of WeChat-based remote follow-up management on reducing the home care burden and anxiety of parents of premature infants. METHODS: A randomized controlled study was conducted in our hospital, with 150 premature infants enrolled in this study, each with 75 cases in the intervention group and the control group. WeChat-based remote follow-up management was performed in the intervention group after discharge, while traditional outpatient follow-up management was performed in the control group. Zarit Burden Interview (ZBI) scale as the primary outcome was used to compare the parental care burden between the two groups. RESULTS: At the 1-month follow-up, the ZBI (37.1 ± 8.3 vs. 54.2 ± 10.5 [mean ± SD], mean difference: 17.1, the 95% confidence interval for the mean difference: [14.07, 20.13], P = 0.016), Family Caregiver Task Inventory (FCTI) scale (23.1 ± 5.2 vs. 33.4 ± 6.7 [mean ± SD], mean difference: 10.3, the 95% confidence interval for the mean difference: [8.38, 12.22], P = 0.023), Self-Rating Anxiety Scale (SAS) (49.6 ± 8.5 vs. 60.2 ± 10.8 [mean ± SD], mean difference: 10.6, the 95% confidence interval for the mean difference: [7.49,13.71], P = 0.021) and Self-Rating Depression Scale (SDS) (48.2 ± 9.5 vs. 58.8 ± 11.2 [mean ± SD], mean difference: 10.6, the 95% confidence interval for the mean difference: [7.28, 13.92], P = 0.019) scores of the intervention group were lower than those of the control group, and the lower scores indicate better outcomes. CONCLUSION: WeChat-based remote follow-up management of premature infants after discharge can effectively improve parents' ability to care and their psychological state and reduce burden of care.

ß-Elemene suppresses the proliferation of human airway granulation fibroblasts via attenuation of TGF-ß/Smad signaling pathway.

BACKGROUND: Benign airway stenosis is easily to recur as a result of hyperplastic airway granulation tissues. Our previous study proved that a Chinese drug, ß-elemene, could inhibit the proliferation of fibroblasts cultured from these tissues, which could decrease the recurrence rate of this disease. METHODS: To find out the mechanism for this effect, we first cultured normal human airway fibroblasts and human airway granulation fibroblasts with different concentrations of ß-elemene for the same time or the same dose of ß-elemene for different times to explore the dose/time-effect relationship between the drug and these cells. Then we used gene microarray to screen out the most important pathway by which the drug influenced human airway granulation fibroblasts. At last, we assessed the condition of this pathway in human airway granulation fibroblasts and verified the effect of this drug on this pathway. RESULTS: ß-Elemene inhibited the proliferation of human airway granulation fibroblasts in a dose but no time-dependent manner and did not affect normal human airway fibroblasts. Affecting the expression of transforming growth factor ß (TGF-ß)/Smad pathway may be the key mechanism for this effect. This pathway was activated in human airway granulation fibroblasts and ß-elemene inhibited it in a dose-dependent manner. This effect was similar to the pathway inhibitor, SB431542, and exogenous TGF-ß could attenuate this effect. CONCLUSION: These results indicated that suppressing TGF-ß/Smad pathway was possibly the key mechanism by which ß-elemene inhibits the proliferation of human airway granulation fibroblasts. This pathway may be a promising target to treat benign airway stenosis.

A novel external catheter fixation method for chemotherapy using inferior epigastric arterial catheterization for cervical cancer.

The aim of this randomized controlled study was to investigate the effects of a novel external catheter fixation method for chemotherapy using inferior epigastric arterial catheterization for cervical cancer.Patients diagnosed with cervical cancer were randomly divided into a control group (n = 32) and a treatment group (n = 33). Patients in the control group underwent a traditional fixation method using a haemostat, elastic band and abdominal bandage. Patients in the treatment group underwent an improved fixation method using an indwelling needle and membrane cover. We used a visual analogue scale (VAS) to evaluate each patient's comfort score and also recorded the incidence of needlestick injury and the length of injection time in each group. The VAS scores measured before and after chemotherapy in the treatment group were lower than in the control group. The incidence of needlestick injury in the treatment group was significantly lower than in the control group. The length of injection time in treatment group was significantly lower than in the control group. Compared with the traditional fixation method, the improved fixation method not only increased patient comfort but also reduced both the risk of needlestick injury and the length of injection time. This improved technique deserves increased clinical use.

Use of subsequent PET/CT in diffuse large B-cell lymphoma patients in complete remission following primary therapy.

Interim 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (I-PET/CT) is a powerful tool for monitoring the response to therapy in diffuse large B-cell lymphoma (DLBCL). This retrospective study aimed to determine when and how to use I-PET/CT in DLBCL. A total of 197 patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) were enrolled between October 2005 and July 2011; PET/CT was performed at the time of diagnosis (PET/CT0), after 2 and 4 cycles of chemotherapy (PET/CT2 and PET/CT4, respectively), and at the end of treatment (F-PET/CT). According to the International Harmonization Project for Response Criteria in Lymphoma, 110 patients had negative PET/CT2 scans, and 87 had positive PET/CT2 scans. The PET/CT2-negative patients had significantly higher 3-year progression-free survival rate (75.8% vs. 38.2%) and 3-year overall survival rate (93.5% vs. 55.6%) than PET/CT2-positive patients. All PET/CT2-negative patients remained negative at PET/CT4, but 3 were positive at F-PET/CT. Among the 87 PET/CT2-positive patients, 57 remained positive at F-PET/CT, and 32 progressed during chemotherapy (15 at PET/CT4 and 17 at F-PET/CT). Comparing PET/CT4 with PET/CT0, 7 patients exhibited progression, and 8 achieved partial remission. Comparing F-PET/CT with PET/CT0, 10 patients exhibited progression, and 7 achieved partial remission. In conclusion, our results indicate that I-PET/CT should be performed after 2 rather than 4 cycles of immunochemotherapy in DLBCL patients. There is a limited role for subsequent PET/CT in the detection of relapse in PET/CT2-negative patients, but repeat PET/CT is required if the PET/CT2 findings are positive.

Qualitative visual trichotomous assessment improves the value of fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography in predicting the prognosis of diffuse large B-cell lymphoma.

INTRODUCTION: Fluorine-18 fluorodeoxyglucose (18 F-FDG) positron emission tomography/computed tomography (PET/CT) is a powerful tool for monitoring the response of diffuse large B-cell lymphoma (DLBCL) to therapy, but the criteria to interpret PET/CT results remain under debate. We investigated the value of post-treatment PET/CT in predicting the prognosis of DLBCL patients when interpreted according to qualitative visual trichotomous assessment (QVTA) criteria compared with the Deauville criteria. METHODS: In this retrospective study, final PET/CT scans of DLBCL patients treated with rituximab-based regimens between October 2005 and November 2010 were interpreted using the Deauville and QVTA criteria. Survival curves were estimated using Kaplan-Meier analysis and compared using the log-rank test. RESULTS: A total of 253 patients were enrolled. The interpretation according to the Deauville criteria revealed that 181 patients had negative PET/CT scan results and 72 had positive results. The 3 year overall survival (OS) rate was significantly higher in patients with negative scan results than in those with positive results (91.6% vs. 57.5%, P<0.001). The 72 patients with positive scan results according to the Deauville criteria were divided into two groups by the interpretation according to the QVTA criteria: 29 had indeterminate results, and 43 had positive results. The 3 year OS rate was significantly higher in patients with indeterminate scan results than in those with positive results (91.2% vs. 33.5%, P<0.001) but was similar between patients with negative and indeterminate scan results (91.6% vs. 91.2%, P=0.921). CONCLUSIONS: Compared with the Deauville criteria, using the QVTA criteria for interpreting post-treatment PET/CT scans of DLBCL patients is likely to reduce the number of false positive results. The QVTA criteria are feasible for therapeutic outcome evaluation and can be used to guide risk-adapted therapy.

Effect of probenecid on pharmacokinetics and tolerability of olmesartan in healthy chinese volunteers.

BACKGROUND: Olmesartan is an angiotensin II receptor antagonist and is effective and well tolerated in the treatment of arterial hypertension. Probenecid is a well-established hypouricemic agent for the treatment of hyperuricemia and gout. OBJECTIVE: The goal of this study was to examine the impact of coadministration of probenecid on the pharmacokinetic parameters and tolerability of olmesartan in healthy volunteers. METHODS: In a randomized, open-label, 2-way crossover study, 12 volunteers received 2 oral treatments (olmesartan alone or olmesartan plus probenecid) separated by 4 days. Blood samples were obtained for a 48-hour pharmacokinetic evaluation after drug administration. Tolerability was assessed by monitoring vital signs and laboratory tests before and after administration of the study drug. RESULTS: Pharmacokinetic parameters were evaluated in 6 male and 6 female healthy volunteers (mean age, 22 [range, 20-25] years]; weight, 56.0 [range, 51.0-60.0] kg). Probenecid coadministration increased olmesartan Css-av, AUC0→∞, and AUC0-48 by 40%, 50%, and 50%, respectively (P = 0.018, 0.000, 0.000, respectively), but there was no statistical significance for Tmax, t1/2, Css-max, and Css-min between olmesartan plus probenecid and olmesartan alone (P = 0.697, 0.053, 0.521, and 0.734, respectively). No serious adverse event (AE) was reported during the study. The proportion of volunteers with AEs in the olmesartan plus probenecid period (5 of 12 [42%]) was higher than that in the olmesartan-alone period (1 of 12 [8%]). All of the AEs during the olmesartan plus probenecid period were abnormal routine urine test results. The AE in olmesartan-alone period was dizziness. All AEs were classified as mild and considered to be at least possibly related to treatment. All volunteers recovered from the AEs by 2 weeks after the end of the study. CONCLUSIONS: Probenecid increases the exposure speed of olmesartan by increasing the AUC0-48, AUC0→∞, and Css-av. The combined treatment of olmesartan medoxomil with probenecid may increase the occurrence of genitourinary side effects. ClinicalTrials.gov identifier: NCT01907373.

The impact of virtual reality on student engagement in the classroom-a critical review of the literature.

Objective: The purpose of this review is to identify the impact of virtual reality (VR) technology on student engagement, specifically cognitive engagement, behavioral engagement, and affective engagement. Methods: A comprehensive search of databases such as Google, Scopus, and Elsevier was conducted to identify English-language articles related to VR and classroom engagement for the period from 2014 to 2023. After systematic screening, 33 articles were finally reviewed. Results: The use of VR in the classroom is expected to improve student engagement and learning outcomes, and is particularly effective for students with learning disabilities. However, introducing VR into middle school education poses several challenges, including difficulties in the education system to keep up with VR developments, increased demands on students' digital literacy, and insufficient proficiency of teachers in using VR. Conclusion: To effectively utilize VR to increase student engagement, we advocate for educational policymakers to provide training and technical support to teachers to ensure that they can fully master and integrate VR to increase student engagement and instructional effectiveness.

[Primary exploration of individual biological boosting target volume for locally advanced nasopharyngeal carcinoma].

OBJECTIVE: To explore the methods of detecting and defining the biological boosting target volume (BBTV) in patients with locally advanced nasopharyngeal carcinoma (NPC). METHODS: Ten patients with locally advanced NPC treated at our center during September to December 2010 were enrolled. The following factors were used to define and contour BBTV, including dose-deficient area locating at skull-base of gross tumor volume in conventional irradiation, hypoxia area detected by (99m)Tc-HL91 Single-photon emission computed tomography/computed tomography (SPET/CT) imaging and tumor-burdened area evaluated by comparing the pre-treatment and 36 Gy irradiation's CT images scanning at the same treatment position. RESULTS: The volume of BBTV was (17 ± 7) cm(3), accounting for 51% ± 19% of gross tumor volume. In BBTV, the volumes and proportion of skull base dose-deficient, hypoxia and tumor-burdened area were (5.8 ± 2.5) cm(3), (7.7 ± 3.6) cm(3), (9.0 ± 5.8) cm(3) and 34.4%, 45.7% and 53.3% respectively. However, the proportion of overlapping volume between three areas was only 33.1%. CONCLUSION: BBTV may be composed of skull base dose-deficient, hypoxia and tumor-burdened volume. It provides a new reference for individualized radiotherapy in locally advanced NPC.

[Expression of B cell activation factor and correlation analysis in patients of systemic lupus erythematosus and its correlation with periodontitis].

PURPOSE: To detect the level of B cell activating factor (B - cell activating factor of the TNF family, BAFF) in the serum of patientes suffering from systemic lupus erythematosus (SLE) with periodontitis, and analyze the relationship between the expression of BAFF with periodontitis and SLE. METHODS: According to the inclusion criteria, patients visiting the Department of Stomatology and Rheumatology, Shengjing Hospital Affiliated to China Medical University were selected, including 19 patients in the periodontitis group(P group), 22 in the systemic lupus erythematosus group (SLE group), 24 in the systemic lupus erythematosus combined with periodontitis group(SLE+P group), and 20 in the healthy control group(H group). The general information, periodontal probing depth (PD), clinical attachment loss (CAL), gingival sulcus bleeding index(SBI) were collected. Serum samples of patients in each group were collected, and BAFF content was determined by Elisa. Rheumatic and immunological indexes of subjects in SLE group and SLE+P group were determined, and the correlation between BAFF content and periodontal indexes was analyzed. SPSS 20.0 software package was used for statistical analysis. RESULTS: CAL in P+SLE group was significantly higher than that in P group(P＜0.05). Serum BAFF concentrations in SLE+P group, SLE group and P group were significantly higher than those in the healthy control group (P＜0.05). Serum BAFF concentration in SLE+P group was significantly higher than that in SLE group(P＜0.05). ESR, SLEDAI and disease duration in SLE+P group were significantly higher than those in SLE group (P＜0.05). The expression level of BAFF in serum was positively correlated with CAL and SBI(P＜0.01). The expression level of BAFF in serum was positively correlated with PD(P＜0.05). There was significant positive correlation between serum BAFF level and duration of disease and hormone use(P＜0.01). Serum BAFF level was positively correlated with C3 (P＜0.05). CONCLUSIONS: SLE has certain correlation with periodontitis, and serum BAFF in SLE patients with periodontitis is significantly increased.BAFF may be associated with the development of SLE and periodontitis.

[Relationship between peripheral blood micronutrients and four kinds of oral mucosal diseases in children: clinical analysis of 217 cases].

PURPOSE: To investigate the relationship between peripheral blood micronutrient levels and 4 kinds of oral mucosal diseases (minor recurrent aphthous ulcer, angular cheilitis, cheilitis and geographic tongue) in children aged 0~14 years. METHODS: One hundred and fifty-two children with oral mucosal lesions (COML) and 65 healthy children (health control group, HC) were included. The clinical data of each group were recorded separately to compare whether there existed differences in the levels of serum water-soluble vitamins (vitamins B1, B2, B3, B5, B6, B7, B9, B12, C), serum fat-soluble vitamins [vitamins A, E, K, 25(OH)D2, 25(OH)D3], zinc and serum calcium. Whether peripheral blood micronutrients were risk factors associated with the onset of COML was analyzed through disorder multiclass logistic regression with SPSS 23.0 software package. RESULTS: Peripheral blood micronutrients differed in children with minor recurrent aphthous ulcers, cheilitis, and geographic tongue (P＜0.05). Compared with HC group, children in minor recurrent aphthous ulcer group had significantly lower levels of vitamin B1, B6, B7, C, A, and 25(OH)D3 (P＜0.05), and relatively higher rates of vitamin B6 (50.00% vs 13.85%), vitamin B7 (36.76% vs 9.23%), 25(OH)D3 (64.71% vs 36.92%) deficiency and vitamin K excess (8.82% vs 0.00%)(P＜0.005). Multiclass logistic regression analysis showed that vitamin B1, vitamin C, vitamin A deficiency, vitamin B5, and vitamin K excess were risk factors for incidence in children with minor recurrent aphthous ulcer, and each element was independent for each other. Compared with HC group, the levels of vitamin B7 and 25(OH)D3 in children with cheilitis were significantly lower(P＜0.05), and the rate of vitamin B7 deficiency was significantly higher (P＜0.005). Multiclass logistic regression analysis showed that vitamin B7 and vitamin A deficiency were risk factors for cheilitis in children, and the two were independent for each other. Compared with the HC group, vitamin K excess rate was significantly higher in children with geographic tongue (7.14% vs 0.00%) (P＜0.005). Multiclass logistic regression analysis showed that vitamin C deficiency and vitamin K excess were risk factors for the incidence of geographic tongue, and the two were independent for each other. Compared with other groups, peripheral blood micronutrients had no correlation with the pathogenesis of angular cheilitis (P＞0.05). CONCLUSIONS: The occurrence of COML is closely related to peripheral blood micronutrient levels, which suggests that children with COML need to monitor vitamin and mineral levels and supplement treatment when necessary.

[Environmental Process, Effects and Risks of Emerging Contaminants in the Estuary-Coastal Environment].

Human activities and global climate change have contributed substantially to the input of land-sourced pollutants into the aquatic environment, especially for emerging or newly identified contaminants, such as microplastics, emerging persistent organic pollutants, pharmaceuticals, and personal care products. The prevalence and toxicity of these emerging pollutants has raised continued concern for the health and safety of the public worldwide. A review of sources, distribution, interfacial transport processes of microplastics, per-and polyfluorinated compounds, antibiotics, and endocrine-disrupting chemicals and factors that influence their environmental behavior in the estuary-coastal environment have been included. The adverse ecological effects and health risks of these emerging contaminants to humans were also reviewed. Lastly, the direction of future research was provided regarding the environmental behavior of multiple emerging pollutants in the coastal environment and the health risks resulting from their interactions, supporting the prevention and control of marine pollution and the healthy development of the marine economy.

[Preliminary study on the correlation between IL-6,IL-33, and IL-10 and periodontitis and rheumatoid arthritis].

PURPOSE: To determine the expression levels of interleukin-6(IL-6), interleukin-33 (IL-33) and interleukin-10 (IL-10) in gingival crevicular fluid in patients with periodontitis and rheumatoid arthritis, and to investigate the correlation between IL-6, IL-33, IL-10 and periodontitis and rheumatoid arthritis. METHODS: According to the inclusion criteria, patients who were admitted to the Department of Stomatology and Rheumatology and Immunology of Shengjing Hospital of China Medical University were selected, including 21 patients with periodontitis (PD group), 21 patients with rheumatoid arthritis (RA group), and 24 patients with periodontitis combined with rheumatoid arthritis (PD+RA group), 24 healthy patients (H group). General information for each subject was recorded including periodontal probing depth (PPD), clinical attachment loss (CAL), and gingival sulcus bleeding index (SBI). Samples of gingival crevicular fluid were collected from each group, and the contents of IL-6, IL-33 and IL-10 in gingival crevicular fluid were determined by ELISA, and the correlation between IL-6, IL-33, IL-10 and periodontal clinical indicators was analyzed. SPSS 20.0 software package was used to analyze the date. RESULTS: The expression level of IL-6 in PD+RA group was significantly higher than that in H group, PD group and RA group (P<0.05). The content of IL-33 in PD group, RA group and PD+RA group was significantly higher than that in H group (P<0.05), while the content of IL-33 in PD+RA group was significantly higher than that in RA group (P<0.05). The expression level of IL-10 in the RA group was significantly higher than that in the H group, the PD group and the PD+RA group (P<0.05). PPD in PD group was positively correlated with the contents of IL-6 and IL-33 (r=0.62, 0.43), SBI, PPD and CAL in PD+RA group were positively correlated with the contents of IL-33 (r=0.69, 0.58, 0.55). CONCLUSIONS: The levels of IL-6 and IL-33 in gingival crevicular fluid of patients with periodontitis accompanied by rheumatoid arthritis are significantly increased, while the contents of IL-10 are significantly decreased, suggesting that IL-6, IL-33 and IL-10 play an important role in the occurrence and development of periodontitis and rheumatoid arthritis.

Correlation of Tim-3 expression with chemokine levels for predicting the prognosis of patients with glioblastoma.

Glioblastoma (GBM) immunotherapy, which blocks the checkpoint inhibitor molecule T cell immunoglobulin domain and mucin domain-3 (Tim-3), has potential therapeutic applications. However, not all patients do benefit from the targeted therapy. This study aimed to explore Tim-3 expression correlated chemokine profiles and immune cell infiltration and investigate their potential as prognostic markers of glioblastoma (GBM) immunotherapy. We analyzed transcriptional data of GBM from TCGA database, to measure Tim-3 expression by R package DESeq2 analysis and observed differentially expressed genes in GBM samples with high Tim-3 expression levels. We also probed the relative gene enrichment pathways. Tim-3 expression was evident in biological processes including the recruitment of immune cells. We also identified some chemokines related to Tim-3 expression. The expression levels of CCL18, CXCL13 and CCL7 were significantly higher in GBM tissues with high Tim-3 expression than in GBM tissues with low Tim-3 expression. In addition, exploring the relationship between immune cell infiltration and Tim-3 expression suggested that Tim-3 expression was positively related to significant immune cell infiltration.

[A novel GATA4 mutation leading to congenital ventricular septal defect].

OBJECTIVE: To identify the GATA4 gene mutation of congenital ventricular septal defect (VSD) and study the molecular mechanism of a novel mutation. METHODS: The clinical data and blood samples from 185 unrelated subjects with congenital VSD were collected and evaluated together with 200 healthy individuals. The coding exons and the flanking intron regions of the GATA4 gene were amplified by PCR and sequenced using the di-deoxynucleotide chain termination approach. The GATA4 gene was cloned and the corresponding mutant was acquired by site directed mutagenesis. The recombinant plasmid expressing GATA4 and the reporter vector expressing enhanced green fluorescence protein (EGFP) driven by the promoter of atrial natrium peptide (ANP) gene were transfected into HeLa cells with Lipofectamine. The effect of mutated GATA4 gene on the transcriptional activity of encoded transcriptional factor was analyzed by reverse transcription (RT)-PCR. RESULTS: A novel heterozygous missense GATA4 mutation, c.191G>A was identified in 1 VSD patient. The mutation leads to glycine to glutamic acid change at amino acid residue 64 (G64E) in the GATA4 protein. Functional analysis showed that GATA4 G64E mutation decreased the transcriptional activity of GATA4 transcriptional factor. CONCLUSION: A novel heterozygous missense GATA4 mutation, G64E, was identified in 1 VSD patient. The mutation might cause VSD by impairing the transcriptional activity of GATA4 transcriptional factor.

[Identification and functional analysis of a KCNA5 mutation responsible for idiopathic atrial fibrillation].

OBJECTIVE: To investigate the molecular mechanism of idiopathic atrial fibrillation (AF) associated with KCNA5 mutation. METHODS: The clinical data and blood samples from 130 unrelated subjects with idiopathic AF were collected and evaluated in contrast to 200 healthy individuals. The coding exons and the flanking introns of KCNA5 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach to identify potential mutations. Multiple alignment of the KCNA5 encoded protein sequences across species was performed. The KCNA5 gene was cloned and the corresponding mutant was acquired by site directed mutagenesis. The recombinant plasmid expressing or tracing KCNA5 was constructed and transfected into COS-7 cells with Lipofectamine, respectively. The effects of mutated KCNA5 gene on the electrophysiological characteristics and subcellular location of encoded ion channel were explored by patch-clamp and confocal microscope, respectively. RESULTS: A heterozygous missense KCNA5 mutation, c.1580C > T was identified in 1 of 130 idiopathic AF patients. Namely, the triplet substitution of ATG for ACG at codon 527, predicting the conversion of threonine into methionine at amino acid residue 527 (T527M), was detected. Functional analysis revealed that KCNA5 T527M mutation exerted predominant negative effect on the currents but no effect on the subcellular location of encoded ion channel. CONCLUSION: The heterozygous KCNA5 T527M mutation identified in 1 idiopathic AF patient exerts predominant negative effect on the currents of encoded ion channel, thereby conducting to idiopathic AF.

[Novel GATA4 mutations identified in patients with congenital atrial septal defects].

OBJECTIVE: To identify the genetic defects in patients with congenital atrial septal defects (ASD). METHODS: The clinical data and blood samples from 180 unrelated subjects with congenital ASD were collected and evaluated. Two hundred healthy individuals served as controls. The coding exons and the flanking introns of GATA4 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with the sequences publicized in GenBank by the aid of programme BLAST to identify the sequence variations. Clustal W software was applied for analysis of the conservation of altered amino acids. RESULTS: Two novel heterozygous missense GATA4 mutations were identified in 2 out of 180 ASD patients. Namely, the triplet substitutions of GTC for GGC at codon 21 and TCG for CCG at codon 87 were detected, predicting the conversions of glycine into valine at amino acid residue 21 (G21V) and proline into serine at amino acid residue 87 (P87S). None of the two mutations were detected in 200 healthy controls. Across-species alignment of GATA4 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, a single nucleotide polymorphism c.99G>T was observed. However, the polymorphic frequency distribution in ASD cases was similar with that in healthy controls (for genotype GT, χ(2) = 0.7556, P = 0.3847; for allele T, χ(2) = 0.7235, P = 0.3950). CONCLUSIONS: Two novel mutations of GATA4 gene are identified in two unrelated ASD patients. This finding provides new insight into the molecular etiology responsible for ASD.

Prognostic Value of the Modified Systemic Inflammation Score in Patients With Extranodal Natural Killer/T-Cell Lymphoma.

BACKGROUND: Extranodal natural killer/T-cell lymphoma (ENKTL) is a rare and extremely malignant tumor. The systemic inflammation score (SIS), which is based on the pretreatment level of lymphocyte-to-monocyte ratio (LMR) and serum albumin (Alb), has been shown to be of prognostic value in a number of cancers. We integrate several other pretreatment serum inflammatory indicators, including the neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), serum C-reactive protein (CRP) and albumin (Alb) level, to establish a modified systemic inflammatory scoring system to predict clinical outcomes of ENKTL. METHODS: A total of 184 patients with newly diagnosed ENKTL was retrospectively investigated. Systemic inflammatory indexes, including NLR, LMR, CRP, and Alb level were reviewed. Receiver operating characteristic (ROC) curve analysis was carried out to obtain the optimal cut-off value. The associations between cutoff values and overall survival (OS) were analyzed by Kaplan-Meier curves and Cox proportional models. RESULTS: The median age of patients was 44.0 years, ranging from 15 to 82 years. There were 129 (70.1%) male patient. About 57.1% of patients had stage III or IV disease. The optimal cut-off values of NLR and LMR in predicting OS were 3.1 and 2.4, respectively. The clinical standard of CRP and Alb levels at 10 and 40 mg/L, respectively, were chosen as the optimal cut-off values. By multivariate analysis, hemophilic syndrome (hazard ratio [HR]: 10.540, 95% confidence interval [CI]: 3.440-32.291, P < 0.001), advanced Ann Arbor stages (III-IV) (HR: 4.606, 95% CI: 1.661-12.774, P = 0.003), paranasal sinus invasion (HR: 2.323, 95% CI: 1.069-5.047, P = 0.033), NLR ≥ 3.1 (HR: 3.019, 95% CI: 1.317-6.923, P = 0.009), Alb level of <40 mg/L (HR: 0.350, 95% CI: 0.134-0.915, P = 0.032), and radiation therapy (HR: 0.430, 95% CI: 0.205-0.901, P = 0.025) were independent protective factors for ENKTL. We combined two inflammatory indexes NLR and Alb level to establish a modified systemic inflammation score (mSIS). These 184 patients were divided into 3 groups: group 1 (mSIS score of 0), group 2 (mSIS score of 1), and group 3 (mSIS score of 2). The mean OS of these three groups were 42 months (95% CI: 31.4-53.12), 77 months (95% CI: 68.5-87.5), and 89 months (95% CI: 71.4-82.7), respectively (P < 0.001). The Harrell's concordance index (C-index) of mSIS is 0.725. The mSIS could be used to discriminate patients categorized in the low-risk group of International Prognostic Index (IPI) (P < 0.001) and the low-risk and intermediate-risk prognostic index of natural killer cell lymphoma (PINK) group (P = 0.019). CONCLUSION: The pretreatment mSIS could be an independent prognostic factor for OS in patients with ENKTL and warrants further research.

Association of body composition with survival and inflammatory responses in patients with non-metastatic nasopharyngeal cancer.

OBJECTIVES: It is unknown whether or not the body composition is correlated with the prognosis and inflammatory response in patients with nasopharyngeal cancer (NPC). MATERIALS AND METHODS: This cohort included 1767 patients with NPC. Visceral, subcutaneous and intra muscular adipose tissues (VAT, SAT and IMAT), and skeletal muscle index were quantified with computed tomography. We used the optimal stratification to select cut points for VAT, SAT and IMAT. We defined sarcopenia according to a widely used cut-point. The primary endpoint was overall survival (OS). The association between body composition and inflammatory response was also examined. RESULTS: Low VAT, SAT, IMAT and sarcopenia were observed in 260 (14.7%), 451 (25.5%), 773 (43.7%) and 683 (38.7%) patients, respectively. Low VAT (P < 0.001, hazard ratio [HR], 1.884; 95% confidence interval [CI], 1.436-2.473,) and SAT (P = 0.022, HR, 1.334, 95%CI, 1.043-1.706) were both associated worse survival. IMAT and sarcopenia were not with prognostic value. In multivariate analysis, we found the prognostic value of the VAT (HR: 1.544, 95% CI: 1.128-2.114; P = 0.007) was independent of T stage, N stage, disease stage, lactic dehydrogenase, neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), the systemic immune-inflammation index (SII), EBV-DNA and body mass index. We observed higher NLR (P = 0.028) and PLR (P < 0.001) in patients with low SAT. Both low VAT (P = 0.009) and SAT (P = 0.005) were associated with decreased stromal lymphocyte infiltrating intensity. CONCLUSIONS: Among body composition parameters, VAT was an independent prognostic factor, especially in patients with locally advanced NPC.

[Novel NKX2-5 mutations identified in patients with congenital ventricular septal defects].

OBJECTIVE: To identify the novel genetic defects in patients with congenital ventricular septal defect (VSD). METHODS: The clinical data and blood samples from 160 unrelated subjects with congenital VSD were collected and evaluated in comparison with 200 healthy individuals. The coding exons and flanking introns of NKX2-5 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with those publicized in GenBank by the aid of program BLAST to identify the sequence variations. The software Clustal W was applied to analyzing the conservation of altered amino acids. RESULTS: Two novel heterozygous missense NKX2-5 mutations were identified in 3 of 160 VSD patients. The same triplet substitution of TTC for TCC at codon 179, predicting the conversion of serine into phenylalanine at amino acid residue 179 (Ser179Phe), was detected in two cases. Another transition of CGC into AGC at codon 36, leading to the change of arginine into serine at amino acid residue 36 (Arg36Ser), was detected in a third case. These two mutations were not observed in 200 healthy controls. A cross-species alignment of NKX2-5 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, two single nucleotide polymorphisms including a common c. 63A > G and a rare c. 606G > C were observed. However, the polymorphic frequency distributions in VSD cases were not statistically different from those in healthy controls (c. 63A > G: chi(2) = 3.403, P = 0.0651; c. 606G > C: chi(2) = 3.278, P = 0.0702). CONCLUSION: Novel NKX2-5 mutations are identified in patients with ASD. They may provide new insight into the molecular etiology responsible for VSD.