Development and multicenter validation of a novel radiomics-based model for identifying eosinophilic chronic rhinosinusitis with nasal polyps.

BACKGROUND: Reliable noninvasive methods are needed to identify endotypes of chronic rhinosinusitis with nasal polyps (CRSwNP) to facilitate personalized therapy. Previous computed tomography (CT) scoring system has limited and inconsistent performance in identifying eosinophilic CRSwNP. We aimed to develop and validate a radiomics-based model to identify eosinophilic CRSwNP. METHODS: Surgical patients with CRSwNP were recruited from Tongji Hospital and randomly divided into training (n = 232) and internal validation cohort (n = 61). Patients from two additional hospitals served as external validation cohort-1 (n = 84) and cohort-2 (n = 54), respectively. Data were collected from October 2013 to May 2021. Eosinophilic CRSwNP was determined by histological criterion. The least absolute shrinkage and selection operator and the logistic regression (LR) algorithm were used to develop a radiomics model. Univariate and multivariate LR were employed to build models based on CT scores, clinical characteristics, and the combination of radiological and clinical characteristics. Model performance was evaluated by assessing discrimination, calibration, and clinical utility. RESULTS: The radiomics model based on 10 radiomic features achieved an area under the curve (AUC) of 0.815 in the training cohort, significantly better than the CT score model based on ethmoid-to-maxillary sinus score ratio with an AUC of 0.655. The combination of radiomic features and blood eosinophil count had a further improved performance, achieving an AUC of 0.903. The performance of these models was confirmed in all validation cohorts with satisfying predictive calibration and clinical application value. CONCLUSIONS: A CT radiomics-based model is promising to identify eosinophilic CRSwNP. This radiomics-based method may provide novel insights in solving other clinical concerns, such as guiding personalized treatment and predicting prognosis in patients with CRSwNP.

[Clinical analysis of pulmonary nocardiosis associated with bronchiectasis].

Objective: To better understand the clinical characteristics of pulmonary nocardiosis associated with bronchiectasis. Methods: Patients diagnosed as bronchiectasis complicated with pulmonary nocardiosis in 9 tertiary general hospitals in China were enrolled from March 2016 to March 2020, with the record of general data, imaging performance and pathogen. The literature was reviewed. Results: Totally 17 patients were included. There were 12 females and 5 males. The ages ranged from 45 to 79 years, with an average of (63±9) years. There were 15 nonsmokers and 2 smokers, all of whom with chronic course. The clinical manifestations were mostly cough, expectoration, hemoptysis, fever, and dyspnea. The imaging manifestation was bronchiectasis in both lungs, with the most common involvement in the left lower lung, right middle lobe and left lingual lobe. Sputum cultures were positive in 10 cases, bronchoalveolar lavage fluid (BALF) cultures were positive in 6 cases, and next generation gene sequencings were positive in 4 cases, including 2 cases of Nocardia gelsenkii, 2 cases of Nocardia abscess, 2 cases of Nocardia stellate, 1 case of Nocardia mexicana, 1 case of Nocardia otitis caviae, and 9 cases of undetermined Nocardia. There were 3 cases of Klebsiella pneumoniae, 2 cases of Pseudomonas aeruginosa and 2 cases of Aspergillus. The symptoms and imaging of all patients were improved after anti Nocardia therapy. Conclusions: Bronchiectasis combined with nocardiosis is more common in middle-aged and elderly women without smoking, which is similar to the clinical manifestations of Lady Windermere syndrome. Bronchiectasis often involves the left lower lobe, right middle lobe and left lingual lobe. Nocardia infection might further precipitate the initiation and progression of bronchiectasis.

[Association between plasma inflammatory mediators and histological endotypes of nasal polyps].

Objective: To compare the clinical characteristics and plasma inflammatory markers levels in different endotypes of chronic rhinosinusitis with nasal polyps (CRSwNP), and to explore the plasma biomarkers associated with endotypes of CRSwNP. Methods: A total of 74 CRSwNP patients (male/female: 41/33; average age: 40 years) and 40 control subjects underwent septoplasty in Tongji Hospital from January 2015 to December 2017 were enrolled in this study. The demographic and clinical features of all subjects including age, gender, past history, visual analogue scale (VAS) and CT scores were recorded. Patients with CRSwNP were divided into EoshighNeuhigh, EoshighNeulow, EoslowNeuhigh and EoslowNeulow four endotypes according to the eosinophil (Eos) percentage and neutrophil (Neu) count of nasal polyps tissue. Preoperative blood routine was performed and the levels of 27 biomarkers in plasma were measured by Bio-Plex suspension chip method. The clinical characteristics and the level of serum biomarkers of patients with different endotypes were compared. SPSS 18.0 software was used for statistical analysis. Results: There was no difference in the clinical features including gender ratio, age, course of disease, VAS score, endoscopy and CT score among EoshighNeuhigh, EoshighNeulow, EoslowNeuhigh and EoslowNeulow CRSwNP patients. Compared with EoslowNeuhigh and EoslowNeulow CRSwNP patients, patients with EoshighNeuhigh and EoshighNeulow endotype demonstrated a higher prevalence of atopy, allergic rhinitis and asthma comorbidity, and increased peripheral blood eosinophil absolute count and percentage (all P<0.05). However, there was no significant difference between EoshighNeuhigh and EoshighNeulow CRSwNP. Plasma levels of all 27 mediators including type 1 cytokines (IL-12 and IFN-γ), type 2 cytokines (IL-4, IL-5 and IL-13), type 3 cytokines (IL-17A), pro-inflammatory cytokines (IL-6 and TNF-α) and tissue remodeling-related markers (bFGF, VEGF and PDGF-BB) demonstrated no significant difference among all endotypes of CRSwNP (all P>0.05). Conclusions: Eoshigh and Eoslow CRSwNP patients display significant differences regarding the prevalence of atopy, allergic rhinitis and asthma comorbidity, peripheral blood eosinophil absolute count and percentage, but the clinical characteristics, blood cellular and biological markers can not effectively distinguish four endotypes of CRSwNP. Further studies are warranted to dig out the potential objective, convenient and reliable markers associated with endotypes in patients with CRSwNP.

Renal outcomes in Asian patients receiving oral anticoagulants for non-valvular atrial fibrillation.

INTRODUCTION: Patients with non-valvular atrial fibrillation (NVAF) may be prescribed warfarin or a non-vitamin K oral anticoagulant (NOAC). There is increasing evidence that NOACs are superior to warfarin in terms of renal function preservation. This study aimed to compare renal outcomes in Chinese patients with NVAF between patients receiving NOACs and patients receiving warfarin. METHODS: In total, 600 Chinese patients with NVAF receiving oral anticoagulant therapy were retrospectively identified from an administrative database. The renal outcomes (≥30% decline in estimated glomerular filtration rate [eGFR], doubling of serum creatinine, and kidney failure) were compared among four propensity-weighted treatment cohorts (warfarin, n=200; rivaroxaban, n=200; dabigatran, n=100; and apixaban, n=100). RESULTS: The mean follow-up period across all groups was 1000 ± 436 days. Compared with warfarin, the three NOACs (pooled for consideration as a single unit) had significantly lower risks of ≥30% decline in eGFR (hazard ratio [HR]=0.339; 95% confidence interval [CI]=0.276-0.417) and doubling of serum creatinine (HR=0.550; 95% CI=0.387-0.782). Dabigatran and rivaroxaban users both had lower risks of ≥30% decline in eGFR (both P<0.001) and doubling of serum creatinine (both P<0.05). Apixaban was only significantly associated with a lower risk of ≥30% decline in eGFR (P<0.001). CONCLUSIONS: Compared with warfarin, NOACs may be associated with a significantly lower risk of decline in renal function among Chinese patients with NVAF.

[Emulation pulmonary nodules localization model:a novel non-invasive localization technique in resection of pulmonary nodules].

Objective: To explore the accuracy and efficiency of a novel 3D-printed emulation localization model of small pulmonary nodules in lung surgery. Methods: From April 2020 to April 2021, a total of 66 patients were selected in the study, who underwent localization and resection of pulmonary nodules with video-assisted thoracoscopic surgery (VATS) guided by the 3D-printed emulation localization model at Department of Thoracic Surgery, West China Hospital of Sichuan University. There were 13 males and 53 females, aged from 25 to 79 (52.7±11.4) years. Of all patients, 24 (36.4%) had single pulmonary nodule, and 42 (63.6%) had synchronous multiple pulmonary nodules. The chest high-resolution CT image data were utilized for digital reconstruction and 3D printing to make a tailored life-size emulation pulmonary nodules localization model, which was used to navigate real-time intraoperative localization of nodules. Clinical data including operative parameters, localization information, resection types and pathological findings of nodules were analyzed. The pulmonary nodules that doctors planned to resect were categorized into two categories:major nodules and additional nodules, according to their presence of invasion and radiological risk factors. The accuracy of localization and resection efficiency of nodules were evaluated in accordance with the categories of the nodules respectively. Results: On the basis of preoperative evaluation, there were 71 major nodules with median maximal diameter of 0.9 (0.6-1.3) cm, and 77 additional nodules with median maximal diameter of 0.5 (0.4-0.7) cm. All patients underwent VATS surgery, 52 of them (78.8%) were treated with uniportal VATS and 14 (21.2%) with triportal VATS. Among the patients with single nodule, 18 segmentectomies and 6 wedge resections were performed; whereas among the patients with multiple nodules, 5 segmentectomies, 14 wedge resections, and 23 combined pulmonary resections (including 2 cases of lobectomy+segmentectomy, 7 cases of lobectomy+wedge resections, and 14 cases of segmentectomy+wedge resections) were achieved. The median operative time was 93 (45-240) min, and the median resection time for all nodules was 51.4 (6.7-147.0) min. All major nodules were successfully resected and visibly dissected after removal, and all additional nodules were successfully resected with 85.7%(66/77) nodules visibly dissected. The accuracy rate of localization of both types of nodules was 100%. All major nodules were malignant, and the malignancy rate of additional nodules was 21.2%(14/66). Conclusion: This novel 3D-printed emulation localization model of small pulmonary nodules proved to be a non-invasive, accurate and efficient technique. Not only that, it has a unique advantage in localization of synchronous multiple pulmonary nodules.

[Multi-level scanning of environmental factors of body mass index of children and adolescents aged 7-17 years in China].

Objective: To investigate the influence of environmental factors on body mass index of children and adolescents in China. Methods: Using data from the China Health and Nutrition Survey, the research object to 7 - 17 years old children and adolescents who participated in 2000, 2006, 2011, and 2015 round of survey with complete data, a total of 6 626 children and adolescents (male 3 473, female 3 153) were investigated under univariate analysis for each environmental factor variable before using Partial Least Square Regression fitting a linear model for further screening. Finally, we fitted a three-level linear mixed-effects model distinct by urban and rural area for analysis. Results: The three-level null model, log likelihood=-17 034.68, χ2=483.06, P<0.001. Intern-class correlation coefficient (ICC) showed that community-level was 9.97%, and both community and individual were 39.38%. The three-level model also showed that urban model's urbanization index (ß=-0.05, 95%CI: -0.09--0.01, P<0.05), the park location (ß=-0.88, 95%CI: -1.72 - -0.04, P<0.05), 15 - 17 age group (ß=-1.04, 95%CI:-1.78 - -0.30, P<0.05) were negatively correlated with BMI. The distance to the gym (ß=0.12, 95%CI: 0.02 - 0.22, P<0.05), the number of home TV sets (ß=0.50, 95%CI: 0.08 - 0.92, P<0.05) and the frequency of parents' alarm of fiction TV program contents (ß=1.85, 95%CI: 0.70 - 3.00, P<0.05) were correlated with BMI. Rural urbanization index (ß=-0.04, 95%CI:-0.07 - -0.01, P<0.05). Rural per capita income (ß=-7.29e-4, 95%CI:-1.00e-3 - -6.77e-5, P<0.05), parents' restricted frequency of watching TV (ß=-1.29, 95%CI:-2.36 - -0.21, P<0.05), adipo-energy ratio (ß=-0.03, 95%CI:-0.06 - -1.00e-3, P<0.05) were negatively correlated with the BMI. Factors as the frequency of parents' alarm of fiction TV program contents (ß=3.01, 95%CI: 0.03 - 6.00, P<0.05), the survey time was 2015 (ß=4.83, 95%CI: 1.96 - 7.69, P<0.05) were correlated with BMI. Conclusions: Environmental factors could indirectly influence the change of BMI of children and adolescents to different degrees and various aspects. Urbanization index and rural per capita income had a slight protective effect on increasing BMI of children and adolescents. At the community level, attention should be paid to the setting of activity places around the living environment of children and adolescents. Family members should also guide their children and adolescents to develop nice behavior in watching TV.

[Performance of vaginal self-sampling high-risk HPV genotyping as primary and combining cytology or viral load as secondary in cervical cancer screening].

Objective: To evaluate the efficacy of high-risk HPV (HR-HPV) genotyping with vaginal self-sampling in primary screening and combining cytology or viral load for HR-HPV positive as secondary screening strategies. Methods: The data referring to HR-HPV genotyping of self-collected sample with mass array matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), HR-HPV viral load of physician-collected sample with hybrid capture Ⅱ (HC-Ⅱ), liquid-based cytology and histology of 8 556 women were from Shenzhen cervical cancer screening trial Ⅱ (SHENCCAST-Ⅱ) conducted between April 2009 and April 2010. The data were reanalyzed to determine the sensitivity and specificity to cervical intraepithelial neoplasia (CIN) of grade 2 or worse (CIN Ⅱ+), CIN of grade 3 or worse (CIN Ⅲ+) when HR-HPV genotyping combining with colposcopy as primary screening strategy based on varied HR-HPV subtype (strategy 1, including 5 sub-strategies: 1a: HPV 16/18 positive; 1b: HPV 16/18/58 positive; 1c: HPV 16/18/58/31/33 positive; 1d: HPV 16/18/58/31/33/52 positive; 1e: any HR-HPV positive). The data were also compared to determine the efficacy of cytology (strategy 2, including 5 sub-strategies: 2a, 2b, 2c, 2d, 2e) or HR-HPV viral load (strategy 3, including 4 sub-strategies: 3a, 3b, 3c, 3d) of physician-collected sample as a triage with HR-HPV genotyping for self-sampling HR-HPV positives. Results: (1) The HR-HPV positive rate was 13.77% (1 178/8 556) in the self-collected samples of 8 556 pregnant women. Of them,the prevalences of HPV 16/18, HPV 16/18/58, HPV 16/18/58/31/33 and HPV 16/18/58/31/33/52 were 3.16% (270/8 556), 5.14% (440/8 556), 6.66% (570/8 556) and 9.81% (839/8 556), respectively. The HR-HPV viral load ≥10 relative light units/control (RLU/CO) was 8.87%(759/ 8 556), while cytological results ≥atypical squamous cell of undetermined signification (ASCUS) were 12.05% (1 031/8 556). (2) The strategy 1e had the highest sensitivities for CIN Ⅱ+, CIN Ⅲ+ which were 92.70% and 94.33%,respectively,among 14 sub-strategies,while the lowest specificity and positive predictive value (PPV). Meanwhile,the required colposcopy referral rates were much higher than other 13 sub-strategies (13.77%). The other 4 sub-strategies of strategy 1 (1a, 1b, 1c, 1d), strategy 1a had the highest specificities for CIN Ⅱ+ and CIN Ⅲ+ (97.92%, 97.69%, respectively), while 1d had the highest sensitivities for CIN Ⅱ+ and CIN Ⅲ+ (88.41%, 92.20%, respectively). (3) Both strategies of referring self-sampling HPV 16/18 positives for immediate colposcopy followed by triage physician-collected sample cytology (≥ASCUS) or viral load (≥10 RLU/CO) for non-HPV 16/18 positives had significantly higher sensitivity and specificity for CIN Ⅱ, CIN Ⅲ+, as well as lower referral rates (strategy 2a and 3a). Additionally, based on these two secondary screening strategies, cumulatively using the other four HR-HPV (HPV 58, 31, 33 and 52) positives as triage for immediate colposcopy showed an enhanced sensitivity. Conclusions: Primary HR-HPV cervical cancer screening strategy based on self-sampling with triage of cytology (≥ASCUS) or viral load (≥10 RUL/CO) provides a good balance among sensitivity, specificity for CIN Ⅱ+ and CIN Ⅲ+ and the number of tests required, referral rates. The efficacy of HR-HPV genotyping combining cytology or viral load secondary screening strategies will have a spiral escalation when HPV 58, 31, 33, 52 are included.

Predicting difficult-to-treat chronic rhinosinusitis by noninvasive biological markers.

BACKGROUND: Previous studies have been limited to the utility of clinical features and invasive nasal mucosal biomarkers in the prediction of chronic rhinosinusitis (CRS) outcomes. This study aimed to identify noninvasive biomarkers associated with difficult- to-treat CRS, enabling physicians to subgroup patients into risk groups for poor outcome before surgery. METHODS: Three hundred and nine CRS patients undergoing endoscopic sinus surgery were finally enrolled. Patients treated with oral or intranasal glucocorticoids within 3 months or 1 month before surgery, respectively, were excluded. Baseline clinical charac- teristics, nasal secretions and peripheral blood samples were collected before surgery. The protein levels of 39 biological mar- kers were detected by the Bio-Plex suspension chip method. Classification and regression tree analysis was applied to establish prediction model for difficult-to-treat CRS determined one year after surgery. A random forest algorithm was used to confirm the discriminating factors that formed the classification tree. RESULTS: In the cohort with nasal secretion sample (n = 189), 21% of CRS patients were diagnosed as difficult-to-treat after 1 year of follow-up. Nasal secretion CCL17 level, hyposmia score, allergic rhinitis comorbidity, and nasal secretion MIP-1ß level were found important predictors of difficult-to-treat CRS. A classification tree separated patients into 5 subgroups leading to an overall predictive accuracy of 94%. However, none of the plasma biological markers were associated with difficult-to-treat CRS in the cohort with blood sample (n = 128). CONCLUSIONS: Patients with difficult-to-treat-CRS were characterized by higher nasal secretion levels of CCL17 and MIP-1ß severe hyposmia and concomitant allergic rhinitis. The classification tree could be useful to identify patients with high risk of poor outcome prior to surgery and offer more personalized interventions. However, since only patients without preoperative steroid treatments were included in this study, the generalization of our predictive model in other patient populations should be conside- red with caution.

Development of a blocking immunoperoxidase monolayer assay for differentiation between pseudorabies virus-infected and vaccinated animalss.

Pseudorabies (PR) outbreaks have devastated many swine farms in several parts of China since late 2011. The outbreak-associated pseudorabies virus (PRV) variant strains exhibited some typical amino acid changes in glycoprotein E (gE), a diagnostic antigen used for discriminating between PRV-infected and vaccinated animals (DIVA). To counteract the potential impact of epitope variations on current serological diagnostics of PRV, we produced monoclonal antibodies (mAbs) against gE protein of one representative PRV variant strain and developed a blocking immunoperoxidase monolayer assay (b-IPMA) for DIVA. The b-IPMA was based on the inhibition of binding between PRV-infected cells and mAb by PRV-specific antibodies present in clinical swine sera and was validated by comparison with a commercial PRV gpI Antibody Test Kit (IDEXX Laboratories, USA). The diagnostic sensitivity, diagnostic specificity and agreement were determined to be 99.25%, 98.18% and 99.02% respectively upon testing 509 serum samples. b-IPMA detected only PRV-specific antibodies and showed no cross- -reactivity with antibodies elicited by gE-deleted vaccine or other common swine pathogens. Thus, b-IPMA has the potential to be used for high-throughput screening of PRV-infected animals in veterinary clinics.

[Metabolomics analysis identifies serum glycerophospholipid expression: a comparison between men and women with asthma].

Objective: To determine whether asthma has a different metabolic glycerophospholipid profile in serum between women and men with asthma. Methods: Fifty-one outpatients with asthma (17 men, 34 women) were enrolled from Peking University Third Hospital from Jan 2015 to Dec 2015. Clinical data such as gender, age, body mass index, pulmonary function were recorded. Glycerophospholipid profile were measured in serum using liquid chromatography-mass spectrometry (LC-MS)-based metabolomics analysis. Projections to latent structures-discriminate analysis (PLS-DA) was used to compare the differences of glycerophospholipid level between men and women. Receiver Operating Characteristic (ROC) Curve was established from men and women. Results: Significantly different glycerophospholipid level were confirmed quantitatively between women and men. Levels of phosphatidylethanolamine (PE), phosphatidylcholine acetal phospholipid (PCP), lysophosphatidyl ethanolamine (LPE), alkyl phosphatidylethanolamine [PE(O)] were significantly higher in women relative to men [PE36: 2, PCP32: 1, LPE18: 0, and PE(O40: 7) was 0.050 (0.037, 0.079) vs 0.043 (0.000, 0.071), 0.057 (0.035, 0.727) vs 0.034 (0.000, 0.057), 0.233 (0.129, 0.390) vs 0.126 (0.075, 0.212), 0.007(0.000, 0.041) vs 0.000 (0.000, 0.000), respectively, all P<0.05). Levels of lysophosphatidylcholine (LPC), lysophosphatidylserine (LPS), and lysophosphatidylphosphatidylcholine [LPC(O)] were significantly lower in women compared to men [Levels of LPS22: 6, LPS20: 4, and LPS18: 1 were 0.000(0.000, 0.003) vs 0.009(0.000, 0.012), 0.015(0.010, 0.026) vs 0.047(0.022, 0.081), 0.008(0.003, 0.179) vs 0.020(0.008, 0.040), respectively, all P<0.05]. Area Under ROC Curve (AUC) of LPS (LPS20: 4) was 0.814. Conclusions: Glycerophospholipid levels in serum are significantly different between women and men asthmatic patients. LPS may contribute to gender based differences in asthmatics.

[Analysis on cardio-metabolic related risk factors in farmers of 15 provinces in China].

Objective: To analyze the prevalence and co-prevalence of cardio-metabolic related risk factors in farmers aged ≥18 years in China, to explore the influence of population economic factors on them. Methods: A total of 3 367 farmers, including fishermen or hunters, aged ≥18 years were selected as study subjects from the database of Nutritional Status and Health Transition of Chinese Residents Project in 2015. Basic information (age, gender), data on anthropometric (body height, weight and waist size), blood biochemical and socioeconomic (occupation, income, education level and living area) were included. According to the definition of the metabolic syndrome released by the International Diabetes Federation (IDF) in 2005, five cardio-metabolic risk factors appeared as central obesity, increased triglycerides, decreased HDL-C, increased blood pressure and increased plasma glucose. Co-prevalence of risk factors was defined as detecting 2 or more risk factors in a person at the same time. Multivariate logistic regression model was used to analyze the relationship between socioeconomic factors and metabolic risk factors. Results: In 3 367 framers of 15 provinces (autonomous region and municipality), the prevalence rates of central obesity, increased blood pressure, increased plasma glucose, increased triglycerides and decreased HDL-C were 51.8%, 59.0%, 17.0%, 25.5% and 38.7% respectively. Multivariate logistic regression analysis showed that the risks for central obesity (OR=3.69, 95%CI: 3.17-4.28) and decreased HDL-C (OR=3.28, 95%CI: 2.81- 3.82) were higher in women than in men, and the risks for increased blood pressure (OR=0.73, 95%CI: 0.63-0.84), increased blood glucose (OR=0.80, 95%CI: 0.67-0.97) were lower in women than in men. Age was positively correlated with the prevalence or co-prevalence of metabolic risk factors (trend P<0.05). Framers in western China had obviously lower risk for central obesity compared with farmers in central China. No significant correlation was found between farmers' income level, education level or the prevalence of metabolic risk factors. Conclusion: In 15 provinces of China, the prevalence of at least 1 kind of cardio-metabolic risk factor was found in 85.5% of the farmers, and the co-prevalence of cardio-metabolic risk factor was found in 60% of farmers. The prevalence and co-prevalence of cardio-metabolic risk factors were significantly associated with age and gender. It is suggested to take targeted nutritional intervention and health education according to the distribution characteristics of prevalence and co-prevalence of cardio-metabolic factors and strengthen the early prevention and control programs of the diseases.

[Elevated Fas expression is related to increased apoptosis of circulating CD8(+)T cell in patients with hepatocellular carcinoma].

Objective: To investigate the mechanism of apoptosis of CD8(+)T lymphocyte in peripheral blood of patients with hepatocellular carcinoma (HCC). Methods: The proportion and apoptosis of peripheral blood CD8(+)T lymphocytes in 30 healthy controls, 30 patients with cirrhosis and 60 HCC patients were detected by Flow cytometry, and the expression of Fas on the surface of CD8(+)T lymphocytes was reported. The differences between groups were compared using independent sample t-test, and data of variance were tested with Mann-Whitney U non-parametric test, P < 0.05 was considered statistically significant. Results: The proportion of CD8(+)T lymphocytes in peripheral blood of patients with HCC was 26.4% ± 9.2%, higher than that of 24.5% ± 7.1% in cirrhosis (t = 0.783, P = 0.489), and and healthy control 19.7% ± 4.7% (t = 2.920, P = 0.004). The proportion of apoptotic CD8(+)T lymphocytes in peripheral blood of HCC patients was 25.3% ± 6.5%, of the total CD8(+)T lymphocytes, which was significantly higher than that of healthy controls 12.1%±6.5% (t = 7.555, P < 0.001) and cirrhotic 13.6% ± 5.8% (t = 5.213, P < 0.001), the differences were statistically significant. The proportion of Fas(+)CD8(+)T lymphocytes in the HCC group was 62.2% ± 18.5%, higher than that in the healthy control group 42.6%±16.5% (t = 4.127, P < 0.001) and 46.1% ± 14.5% (t = 2.561, P < 0.01)of the cirrhosis group, the differences were statistically significant. Fas expression was positively correlated with the apoptosis of CD8(+)T lymphocytes (r (2) = 0.113, P < 0.05). Conclusion: The proportion of CD8(+)T lymphocytes in peripheral blood of patients with HCC is higher than that of healthy controls, but the proportion of CD8(+)T lymphocyte apoptosis based on Fas/FasL pathway increased, which may be an important mechanism for tumor cell immune escape.

Effects of graded removal of lysine from an intravenously infused amino acid mixture on lactation performance and mammary amino acid metabolism in lactating goats.

To investigate responses of milk protein synthesis and mammary AA metabolism to a graded decrease of postruminal Lys supply, 4 lactating goats fitted with jugular vein, mammary vein, and carotid artery catheters and transonic blood flow detectors on the external pudic artery were used in a 4 × 4 Latin square experiment. Goats were fasted for 24 h and then received a 9-h intravenous infusion of an AA mixture plus glucose. Milk yield was recorded and samples were taken in h 2 to 8 of the infusion period; a mammary biopsy was performed in the last hour. Treatments were graded decrease of lysine content in the infusate to 100 (complete), 60, 30, or 0% as in casein. Lysine-removed infusions linearly decreased milk yield, tended to decrease lactose yield, and tended to increase milk fat to protein ratio. Milk protein content and yield were linearly decreased by graded Lys deficiency. Mammary Lys uptake was concomitantly decreased, but linear regression analysis found no significant relationship between mammary Lys uptake and milk protein yield. Treatments had no effects on phosphorylation levels of the downstream proteins measured in the mammalian target or rapamycin pathway except for a tended quadratic effect on that of eukaryotic initiation factor 2, which was increased and then decreased by graded Lys deficiency. Removal of Lys from the infusate linearly increased circulating glucagon and glucose. Removal of Lys from the infusate linearly decreased arterial and venous concentrations of Lys. Treatments also had a significant quadratic effect on venous Lys, suggesting mechanisms to stabilize circulating Lys at a certain range. The 2 infusions partially removing Lys resulted in a similar 20% decrease, whereas the 0% Lys infusion resulted in an abrupt 70% decrease in mammary Lys uptake compared with that of the full-AA mixture infusion. Consistent with the abrupt decrease, mammary Lys uptake-to-output ratio decreased from 2.2 to 0.92, suggesting catabolism of Lys in the mammary gland could be completely prevented when the animal faced severe Lys deficiency. Mammary blood flow was linearly increased, consistent with the linearly increased circulating nitric oxide by graded Lys deficiency, indicating mechanisms to ensure the priority of the mammary gland in acquiring AA for milk protein synthesis. Infusions with Lys removed increased mammary clearance rate of Lys numerically by 2 to 3 fold. In conclusion, the decreased milk protein yield by graded Lys deficiency was mainly a result of the varied physiological status, as indicated by the elevated circulating glucagon and glucose, rather than a result of the decreased mammary Lys uptake or depressed signals in the mTOR pathway. Mechanisms of Lys deficiency to promote glucagon secretion and mammary blood flow and glucagon to depress milk protein synthesis need to be clarified by future studies.

[Characteristics of IgH-CDR3 repertoire of peripheral B cells in a patient with primary biliary cholangitis: a preliminary study using high-throughput sequencing].

Objective: To analyze the characteristics of immunoglobulin heavy chain complementarity-determining region (IgH-CDR3) repertoire of peripheral B cells in a patient with primary biliary cholangitis (PBC) and to investigate the diversity of the immune system. Methods: Arm-PCR was used to amplify the IgH-CDR3 region of circulating B cells isolated from a PBC patient, and high-throughput sequencing was used to analyze the amplified product. The characteristics of immune repertoire were analyzed by bioinformatics. Results: In total, 329219 sequence reads were generated from the sample, with 325540 total CDR3 sequences and 72774 distinct CDR3 sequences, and the D50 of IGH-CDR3 was 7.7. The dominant CDR3 length of the sample was 45 nt (9.6%); the N addition with the highest frequency ranged from 13 to 14 nt (5.25%); the J trimming with the highest frequency was 0 nt (12.7%); the three most frequent V alleles were V4-59 (9.5%), V3-23 (8.1%), and V1-69 (6.4%). Conclusion: The diversity of IgH-CDR3 repertoire is relatively low in this patient with PBC, with several B-cell clonal expansions. The specificity needs to be further verified after increasing the sample size.

Distribution and diversity of bacterial communities and sulphate-reducing bacteria in a paddy soil irrigated with acid mine drainage.

AIMS: To investigate the effects of long-term acid mine drainage (AMD) irrigation on the change in bacterial community and sulphate-reducing bacteria (SRB) in a paddy soil. METHODS AND RESULTS: The bacterial community structures were investigated using 454 pyrosequencing, and 98 931 effective sequences were selected for the bacterial diversity analysis. The known dominant phyla in the paddy soil were Acidobacteria (33·5%), Proteobacteria (19·7%), Nitrospira (2·8%) and Actinobacteria (2·7%). Higher percentage of Acidobacteria than Proteobacteria was detected. The relative abundances of the dominant bacterial lineages were more significantly correlated with the soil pH, the organic matter and the sulphate than the heavy metals. The diversity of the SRB in the surface paddy soil showed that the uncultured SRB groups might play important roles in paddy soils. The other OTUs mainly belonged to six phylogenetic divisions: Desulfobacca, Desulfovibrio, Syntrophobacter, Desulforhopalus, Desulfarculus and Desulfobulbus. The distribution of the absolute abundance and the relative contribution of the SRB along the vertical soil profile were investigated by RT-PCR assays based on the dsrB gene. The abundance of the dsrB gene copy numbers was up to 1·92 × 10(9)  copies g(-1) dry soil, which is slightly higher than the other non-AMD-affected paddy soil. CONCLUSIONS: This study demonstrated that the abundance of SRB is increased by the AMD irrigation while changing the composition and diversity of the bacterial community in the paddy soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This is, to our knowledge, the first attempt to characterize and quantify changes in the diversity and distribution of the microbial community and SRB in the long-term AMD-irrigated paddy soil, which will further increase our understanding of the impact of AMD on sulphur biogeochemical cycling in the paddy soil.