Objective To establish a five-color flow cytometric immunophenotyping protocol and valuate its clinical application in leukemia and lymphoma. Methods Samples from 73 cases of acute leukemia and 30 cases of lymphoma were analyzed using a comprehensive antibody panel with five-color combination. The antibody combination CD7/CD33/CD19/CD13/CD45 and MPO/cCD79a/cCD3/CD117/CD45,composed of different lineage distinctive and lineage sensitive markers of B, T and myeloid cells, were applied to determine the lineage originality of leukemia and lymphoma celia. The markers used in the second step analysis were based on the findings of the first step. Results Five-color compensation can be performed automatically and easily with the advanced digital compensation program. The results showed that the expression ratio of CD19 in B-ALL was 100% (27/27), cCD79, pesitivity was 92.6% (25/27) and CD7,cCD3 was expressed in all T-ALL cases (8/8). The B-ALL could be staged depending on the expression level of CD34 ,CD10 ,cμ,sIgM and the T-ALL could be dearly staged dependent on the expression level of CD2,CD1a,,CD4,CD4,CD8. The expression ratio of cMPO, CD117 was 89. 5% (34/38) and 81.6% (31/38) in AML respectively, however CD7 was also expressed in 26.3% (10/38) of AML cases. Combined with morphology, immanophenotypes could be used for diagnosing AML with subtyping. The CD19/SSC gating can be used for immunophenotyping of B cell lymphoma with differential diagnosis. The number of robes required was significantly reduced with our panel from 12 tubes of 3-color to 6 tubes of 5-color. Implementation of the five-color protocol had resulted in 20% reduction in reagent costs. Conclusions The application of fivecolor staining protocol significantly improve the sensitivity and accuracy of measurement of immunophenotypes of acute leukemia and lymphoma. It reduces the cost and can be widely applied in the clinical laboratory diagnosis.

Objective To investigate the effects of abdominal breathing training on sleep disorders in elderly patients with chronic heart failure. Methods Total of 100 patients with chronic heart failure complain of sleeping disorders and Pittsburgh Sleep Quality Index (PSQI)>7 points were assigned into two groups by random digits table method, 50 cases in each group. The observation group and the control group were nursed in the same way except that abdominal breathing was adapted to the observation group. Sleep status, heart rate, blood pressure, SpO2 and brain natriuretic peptide (BNP) were evaluated before training, one week and eight weeks after training respectively. Statistics was used to analyze the differences between two groups. Results After training one week, the sleep status of the observation group was ameliorated, but without significant difference compared to the control group (P>0.05). And after training eight weeks, the PSQI, BNP and heart rate were (9.21 ± 6.38) points, (193.78 ± 152.16) μg/L, (63.5 ± 10.8) times/min in the observation group, and (12.92 ± 0.33) points, (417.55 ± 262.47) μg/L, (70.7 ± 8.5) times/min in the control group, and there was significant differences between 2 groups (t=3.627, 2.041, 2.767, all P < 0.05), while the blood pressure, SpO2 did not change obviously(P>0.05). Conclusions Abdominal breathing training could ameliorate sleep status in elderly patients with chronic heart failure.

Objective: To investigate the risk factors of ventricular arrhythmias in patients with Brugada syndrome. Methods: Clinical data of 60 Brugada syndrome patients admitted in the department of cardiology of the First Affiliated Hospital of Nanjing Medical University from March 2003 to December 2016 were collected and retrospectively analyzed. The age at diagnosis was (43.2±13.1) years (0.6-83.0 years), 98.3% were males (n=59), and the patients were followed up to (92±41) months (12-169 months). The 12-lead surface electrocardiogram (ECG) recorded at the time of diagnosis and showing the highest type 1 ST elevation, either spontaneously or after provocative drug test, was used for the analysis. Patients were divided into ventricular arrhythmia (VA, n=12) group and non-ventricular arrhythmia (non-VA, n=48) group depending on the presence or absence of clinical VA event. The demographic data and ECG data of the 2 groups were compared, and the independent risk factors of VA events were analyzed by stepwise logistic regression. Results: Incidence of family history of sudden death (7/12 vs. 22.9% (11/48)) and percentage of type 1 ST elevation in the peripheral ECG leads (6/12 vs. 16.67% (8/48)) were significantly higher in VA group than in non-VA group (both P<0.05). Max Tpeak-Tend (Max-Tpe) interval ((144±53)ms vs. (110±16)ms) and dispersion of Tpe ((74±50)ms vs. (43±17)ms) were significantly higher in VA group than in non-VA group (both P<0.05). The area under receiver operating characteristic (ROC) curves for the Max-Tpe interval was 0.693 and Max-Tpe interval ≥140 ms was determined as an optimized cutoff point with increased risk of VA event, which had a sensitivity of 50.0%, a specificity of 98.0%, a positive predictive value of 85.7%, and a negative predictive value of 88.7% for predicting VA event. The ROC curves for the dispersion of Tpe was 0.775 and dispersion of Tpe ≥45 ms was determined as an optimized cutoff point for predicting VA event, which had a sensitivity of 91.7%, a specificity of 64.6%, a positive predictive value of 39.3%, and a negative predictive value of 96.9% for predicting VA event. In multivariate analysis, Max-Tpe interval ≥140 ms (OR=27.53, 95%CI 1.07-706.77, P=0.045) and family history of sudden death (OR=24.63, 95%CI 2.05-295.38, P=0.011) were found to be the independent risk factors of arrhythmic events. Conclusions Max-Tpe interval ≥140 ms and family history of sudden death are risk factors of VA event in included patients with Brugada syndrome.

Objective To investigate changes in gait level of patients after hip replacement, the variation trend of bone mineral density (BMD) around the prothesis was studied, so as to reveal the influence pattern of gait level at postoperative initial and long-term stages on bone reomodeling. Methods Based on adaptive bone remodeling theory, the finite element model of femer-prosthesis was developed. The BMD distribution was calculated using the initial and long-term gait level after hip replacement as the remodeling parameters. Gruen method was applied to quantify the BMD changes. Results At the postoperative initial stage, obvious variations existed in constant gait group and changing gait group. The maximum difference occurred in low gait group, resulting in the decrease of BMD by 41% in greater trochanter region. The improvement of gait level would promote the enhancement of BMD in proximal and middle region of the prosthesis, resulting in the increase of BMD by 47%. Long-term gait recovery would promote BMD recovery in middle and end region of prosthesis, with BMD increase by 2%-9%. Conclusions The research findings provide guidance for rehabilitation process of patients after hip replacement.

ObjectiveTo investigate the risk factors for portopulmonary hypertension （POPH） in liver cirrhosis， and to construct a noninvasive predictive model. MethodsA retrospective analysis was performed for the clinical data of 310 cirrhotic patients with portal hypertension who were hospitalized in The Third Affiliated Hospital of Hebei Medical University from January 2013 to August 2022， and according to whether pulmonary artery systolic pressure was ≥40 mmHg on ultrasound， the patients were divided into POPH group with 31 patients and non-POPH group with 279 patients. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. A binary Logistic regression analysis was used to determine the independent risk factors for POPH， and a nomogram prediction model was constructed. The Bootstrap resampling method was used for internal validation， and C-index and calibration curve were used to assess the discriminatory ability and consistency of the model. The rms package was used to plot the nomogram. ResultsCompared with the non-POPH group， the POPH group had a significantly younger age， a significantly higher proportion of women or patients with hepatic encephalopathy or Child-Pugh class C disease， and significantly higher levels of direct bilirubin， Model for End-Stage Liver Disease score， albumin-bilirubin （ALBI） score， international normalized ratio， prothrombin time， FIB-4 index， LOK score， and Forns index， as well as significantly lower levels of serum albumin， alanine aminotransferase， gamma-glutamyl transpeptidase， hemoglobin， total cholesterol， and triglycerides （all P<0.05）. The multivariate analysis showed that sex （odds ratio ［OR］=0.172， 95% confidence interval ［CI］： 0.064‍ ‍—‍ ‍0.462， P<0.001）， age （OR=0.944， 95%CI： 0.901‍ ‍—‍ ‍0.989， P=0.016）， ALBI score （OR=3.091， 95%CI： 1.100‍ ‍—‍ ‍8.687， P=0.032）， and hepatic encephalopathy （OR=3.466， 95%CI： 1.331‍ ‍—‍ ‍9.031， P=0.011） were independent risk factors for POPH. A predictive model for POPH in liver cirrhosis was established based on the above independent risk factors， with a C-index of 0.796 （95%CI： 0.701‍ ‍—‍ ‍0.890）， suggesting that the model had good discriminatory ability， and the calibration curve showed that the model had good calibration ability， suggesting that the model had certain predictive efficacy. ConclusionYoung female individuals， elevated ALBI score， and comorbidity with hepatic encephalopathy are independent risk factors for POPH in patients with liver cirrhosis， and the predictive model established based on these factors has a certain clinical application value.

Although genome-wide association studies have identified more than eighty genetic variants associated with non-small cell lung cancer (NSCLC) risk, biological mechanisms of these variants remain largely unknown. By integrating a large-scale genotype data of 15 581 lung adenocarcinoma (AD) cases, 8350 squamous cell carcinoma (SqCC) cases, and 27 355 controls, as well as multiple transcriptome and epigenomic databases, we conducted histology-specific meta-analyses and functional annotations of both reported and novel susceptibility variants. We identified 3064 credible risk variants for NSCLC, which were overrepresented in enhancer-like and promoter-like histone modification peaks as well as DNase I hypersensitive sites. Transcription factor enrichment analysis revealed that USF1 was AD-specific while CREB1 was SqCC-specific. Functional annotation and gene-based analysis implicated 894 target genes, including 274 specifics for AD and 123 for SqCC, which were overrepresented in somatic driver genes (ER = 1.95, P = 0.005). Pathway enrichment analysis and Gene-Set Enrichment Analysis revealed that AD genes were primarily involved in immune-related pathways, while SqCC genes were homologous recombination deficiency related. Our results illustrate the molecular basis of both well-studied and new susceptibility loci of NSCLC, providing not only novel insights into the genetic heterogeneity between AD and SqCC but also a set of plausible gene targets for post-GWAS functional experiments.
Adenocarcinoma of Lung/genetics* ; Carcinoma, Non-Small-Cell Lung/genetics* ; Carcinoma, Squamous Cell/genetics* ; Genetic Heterogeneity ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Lung Neoplasms/genetics* ; Polymorphism, Single Nucleotide