pH-dependent water permeability switching and its memory in MoS2 membranes.

Intelligent transport of molecular species across different barriers is critical for various biological functions and is achieved through the unique properties of biological membranes1-4. Two essential features of intelligent transport are the ability to (1) adapt to different external and internal conditions and (2) memorize the previous state5. In biological systems, the most common form of such intelligence is expressed as hysteresis6. Despite numerous advances made over previous decades on smart membranes, it remains a challenge to create a synthetic membrane with stable hysteretic behaviour for molecular transport7-11. Here we demonstrate the memory effects and stimuli-regulated transport of molecules through an intelligent, phase-changing MoS2 membrane in response to external pH. We show that water and ion permeation through 1T' MoS2 membranes follows a pH-dependent hysteresis with a permeation rate that switches by a few orders of magnitude. We establish that this phenomenon is unique to the 1T' phase of MoS2, due to the presence of surface charge and exchangeable ions on the surface. We further demonstrate the potential application of this phenomenon in autonomous wound infection monitoring and pH-dependent nanofiltration. Our work deepens understanding of the mechanism of water transport at the nanoscale and opens an avenue for the development of intelligent membranes.

Ambient air pollution exposure and telomere length: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to provide evidence of the associations between pre- and post-birth and adulthood air pollution exposure with telomere length. STUDY DESIGN: The databases of PubMed, Embase, and Web of Science were searched up to June 1st, 2022 in order to include relevant observational studies and perform a systematic review and meta-analysis. METHODS: The random-effects meta-analysis was grouped by air pollutant and exposure window (pre- and post-birth and adulthood) to evaluate the summary effect estimate. Cochran's Q and I2 statistics were used to evaluate the heterogeneity among the included studies. The quality of individual studies was evaluated using the national toxicology program/office of health assessment and translation risk of bias rating tool. RESULTS: We identified 18 studies, covering 8506 children and 2263 adults from multiple countries. We found moderate evidence that particulate matter less than 2.5 µm (PM2.5) exposure during the entire pregnancy (-0.043, 95% CI: -0.067, -0.018), nitrogen dioxide (NO2) exposure during the first trimester (-0.016, 95% confidence interval [CI]: -0.027, -0.005), long-term adulthood PM2.5 exposure were associated with shortening telomere length. Mild to high between-study heterogeneity was observed for the most tested air pollutant-telomere length combinations in different exposure windows. CONCLUSIONS: This systematic review and meta-analysis provides the evidence which strongly supports that prenatal PM2.5 and NO2 exposures were related to reduced telomere length, while prenatal sulfur dioxide (SO2) and carbon monoxide (CO) exposures, childhood PM2.5, particulate matter less than 10 µm (PM10), NO2 exposures and short-term adulthood PM2.5 and PM10 exposures were not associated with telomere length. Further high-quality studies are needed to elaborate our suggestive associations.

[Intervention effect of apocynin on silicosis induced by silica in rats].

Objective: To investigate the intervention effect and its mechanism of apocynin, an inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) on silicosis induced by silica (SiO(2)) in rats. Methods: In October 2021, 24 SPF SD male rats were divided into control group, silicosis model group and apocynin intervention group according to random number table method, with 8 rats in each group. SiO(2) was exposed by one-time intratracheal instillation. The rats in the apocynin intervention group were intraperitoneally injected with apocynin 50 mg/kg, 3 times a week, on the second day after treatment. The rats were sacrificed 28 days later, and lung coefficients were calculated after lung tissues were weighed. Hematoxylin-eosin staining and Masson staining were used to observe the lung histopathological changes in each group, respectively. The levels of NOX, reactive oxygen species (ROS), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in lung tissue were detected. The expressions of interleukin-1 beta (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by Enzyme-Linked Immunosorbent Assay (ELISA). The level of hydroxyproline (HYP) was detected by alkaline hydrolysate. The expressions of transforming growth factor beta 1 (TGF-ß1), E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) in lung tissue were detected by Western blotting. Results: Compared with the control group, the body weight of silicosis model group was decreased, the lung tissue showed obvious inflammatory infiltration and fibrosis, and the levels of lung coefficient, IL-1ß, IL-6, TNF-α and TGF-ß1 were significantly increased (P<0.05). Compared with the silicosis model group, the lung tissue injury in the apocynin intervention group was significantly improved, the lung coefficient, NOX, ROS, MDA, IL-1ß, IL-6, TNF-α and TGF-ß1 levels were decreased, and the activity of GSH-Px was increased (P<0.05). Compared with the silicosis model group, the expressions of HYP and α-SMA were decreased and the level of E-cad was increased in the apocynin intervention group (P<0.05) . Conclusion: Apocynin may alleviate SiO(2)-induced fibrosis in silicosis rats by reducing oxidative stress, the release of inflammatory factors and inhibiting the process of epithelial-mesenchymal transition.

[An epigenetic clock model for assessing the human biological age of healthy aging].

Objective: To construct an epigenetic clock model for assessing and calibrating human biological age. Methods: Convenience sampling was used to select 186 subjects from the longevity cohort of Guangxi Zhuang Antonornous Region from July 1 to November 30, 2019, and 124 subjects from the physical examination population of the Seventh Medical Center of the PLA General Hospital from October 1 to December 31, 2020. Self-designed questionnaire was applied to collect demographic characteristics and family history of disease. Physical examination was applied to determine heart rate and blood pressure. Fasting peripheral venous blood was drawn for determination of fasting plasma glucose, plasma total cholesterol, triglyceride, plasma high-density lipoprotein cholesterol, plasma low-density lipoprotein cholesterol and telomere length. Methylation levels of EDARADD cg09809672, IPO8 cg19722847, NHLRC1 cg22736354, P2RX6 cg05442902 and SCGN cg06493994 were detected by targeted methylation site sequencing. A total of 54 subjects with unqualified quality control of DNA methylation and telomere length were excluded, and 256 subjects' data were finally analyzed. Trend test was used for the change of methylation level among different ages groups, multiple linear regression method was used to build prediction models of biological age. Kendal rank correlation analysis was used to evaluate the correlation of age gap (Gregorian calendar age minus biological age) with telomere length. Independent sample t-test was used to compare the health-related indicators between subjects with different age gap within different age groups. Results: The M(Q1, Q3)of age of subjects were 67 (51, 91) years old, including 166 females (64.84%). With increase of age, the methylation levels of gene loci were decreased (EDARADD cg09809672, IPO8 cg19722847 and P2RX6 cg05442902) and increased (NHLRC1 cg22736354 and SCGN cg06493994) (all P values<0.05). The established biological age prediction model was as follows: Y=-53.121×EDARADD cg09809672-137.564×IPO8 cg19722847+141.040×NHLRC1 cg22736354-67.893×P2RX6 cg05442902+149.547×SCGNcg06493994+4.592×sex+64.185 (R2=0.86, P<0.001), where Y was the biological age, and the items in the equation were methylation level, sex (male =1, female =2) and intercept in sequence. The Kendall rank correlation coefficient between age gap and telomere length was 0.731 (P<0.001). Compared with the subjects whose age gaP<0, the subjects with age gaP≥0 had higher systolic blood pressure in adolescence [(88.50±8.89) and (109.83±9.48) mmHg, respectively, 1 mmHg=0.133 kPa]; lower TC [(5.48±0.23) and (3.98±0.54) mmol/L, respectively, ] and TG [(3.51±0.32) and (3.41±0.20) mmol/L] in young adults; lower fasting blood glucose in middle age [(6.17±0.67) and (5.37±0.79) mmol/L, respectively, ] and higher diastolic blood pressure in nonagenarian age [(76.99±6.78) and (83.97±9.36) mmHg, respectively, ] (all P values<0.05). Conclusion: The constructed epigenetic clock model can be used to evaluate and calibrate human biological age.

[Assessment of cognitive function of the elderly by serum metabolites of brain-gut axis].

Objective: To investigate the feasibility of assessing cognitive function of the elderly by serum metabolites of brain-gut axis. Methods: Convenience sampling was used to select 100 and 60 participants from the healthy population cohort and microecological balance cohort of the longevity population in Guangxi, to constitute subset of healthy population and longevity population, respectively. A questionnaire was used to investigate the demographic characteristics of the subjects, 2-5 ml of fasting venous blood was collected from the subjects, and the serum untargeted metabolomics was determined by liquid chromatography tandem mass spectrometry. The biomarkers related to the brain-gut axis were collected through literature retrieval, and the results were intersected with the untargeted metabolites and annotated. Spearman correlation analysis was used to screen serum metabolites of brain-gut axis associated with aging, and multiple linear regression method was used to construct biological age model. The mini mental status examination was used to evaluate the cognitive function of longevity population subsets. The differences of biological age and chronological age of longevity population subsets with different cognitive function were compared. Results: The M (Q1, Q3) of subset of healthy population and longevity population were 64 (38, 72) and 97 (95, 99) years old, respectively, and there were 50 (50.0%) and 44 (73.3%) females, respectively. Nine serum metabolites of brain-gut axis were obtained by initial screening, which were propionic acid, glutamic acid, γ-aminobutyric acid (GABA), lactic acid, 5-hydroxytryptamine (5-HT), tryptophan, trimethylamine oxide, dopamine and canine urea. Spearman correlation analysis showed that glutamic acid and dopamine were positively correlated with aging (r values were 0.208 and 0.524, respectively, all P values<0.05), and tryptophan, 5-HT and GABA were negatively correlated with aging (r values were -0.308, -0.533 and -0.213, respectively, all P values<0.05). The biological age model was constructed as: y=49.81-1.18×10-5× GABA-1.82×10-4×5-HT+1.99×10-3×dopamine+1.65×10-6×glutamic acid -2.04×10-6×tryptophan+2.36×gender, where y was the biological age (years), the items on the right were the intercept item, the relative concentration of each metabolite, and gender (male=1, female=2). The coefficient of determination of model was 0.50 (P<0.001). The M (Q1, Q3) of the chronological age of the subset of longevity population with poor, moderate and good cognitive function were 97 (94, 100), 97 (93, 101) and 96 (94, 101) years old, respectively, and there was no statistical significance in pairwise comparison (all P values>0.05). The M (Q1, Q3) of the biological age of the subjects with better cognitive function was 51 (38, 54) years old, which was lower than that of the subjects with poor cognitive function [57 (47, 61)] (P=0.040). Conclusion: The biological age model can be constructed based on serum metabolites of brain-gut axis and used to evaluate the cognitive function of the elderly.

Robotic rectal cancer surgery: comparative study of the impact of obesity on early outcomes.

The aim of this study was to compare the outcomes of robotic total mesorectal excision (TME) in obese versus non-obese patients. A total of 533 patients, of whom 161 were obese (30·2 per cent) underwent robotic proctectomy during the study interval. Patient obesity was not associated with adverse short-term clinical outcomes after robotic rectal cancer surgery. Indicated in the obese perhaps?

[Correlation between breast density and age, breast cancer molecular subtype].

Objective: To explore the correlation between breast density and breast cancer molecular subtype. Methods: The data of 1 407 cases of breast cancer immunohistochemistry and preoperative mammography in Tianjin Medical University Cancer Institute and Hospital from August 2015 to August 2016 were retrospectively analyzed. All cases were divided into four groups: <45 years old, 45-54 years old, 55-64 years old and ≥ 65 years old, and all groups were divided into four molecular subtypes: Luminal A, Luminal B, HER2-enriched and triple-negative according to immunohistochemical results. Person correlation test was used to analyze the correlation between breast density and age; Kruskal-Wallis test was used to analyze the difference in breast density between different molecular subtypes. Results: There was a negative correlation between breast density and age (r=-0.55, P<0.01). Descriptive analysis results showed that the mean breast density of<45 years old, 45-54 years old, 55-64 years old and ≥65 years old was 20.0%±7.3%, 16.2%±8.4%, 10.5%±5.2%, and 7.9%±3.2%, respectively. The mean breast density of Luminal A, Luminal B, HER2-enriched and triple-negative was 14.0%±8.1%, 14.5%±7.6%, 14.8%±7.7% and 13.2%±7.3%, respectively. Kruskal-Wallis test showed significant statistical differences in breast density among molecular subtypes in the group of<45 years old (P<0.05), while no statistical significance in other groups (all P>0.05). Conclusion: There is correlation between breast density and breast cancer molecular subtype in young patients, but whether breast density has a potential evaluation effect on breast cancer molecular subtype still needs to be further studied.

Impact of an intervention programme on knowledge, attitudes and practices of population regarding severe fever with thrombocytopenia syndrome in endemic areas of Lu'an, China.

Knowledge, attitudes and practices (KAP) of the population regarding severe fever with thrombocytopenia syndrome (SFTS) in endemic areas of Lu'an in China were assessed before and after an intervention programme. The pre-intervention phase was conducted using a sample of 425 participants from the 12 selected villages with the highest rates of endemic SFTS infection. A predesigned interview questionnaire was used to assess KAP. Subsequently, an intervention programme was designed and applied in the selected villages. KAP was re-assessed for each population in the selected villages using the same interview questionnaire. Following 2 months of the programme, 339 participants had completed the re-assessed survey. The impact of the intervention programme was evaluated using suitable statistical methods. A significant increase in the KAP and total KAP scores was noted following the intervention programme, whereas the proportion of correct knowledge, the positive attitudes and the effective practices toward SFTS of respondents increased significantly. The intervention programme was effective in improving KAP level of SFTS in populations that were resident in endemic areas.

Limb-salvage treatment of malignant tumors of the limbs.

The aim of this study was to explore the clinical efficacy of limb-salvage therapy for malignant bone tumors of the limbs using two surgical methods. This investigation presents a retrospective study of malignant bone tumors of the limbs in 42 patients. Participants were divided into two groups: group A of 25 patients who were treated with artificial prosthesis replacement, and group B of 17 patients treated with bone inactivation. By collection of clinical data, the survival rate, surgical complications, quality of life, pain relief and postoperative limb function following artificial prosthesis replacement and tumor inactivation were comprehensively evaluated in patients with malignant bone tumors of the limbs. Group A had significantly higher Karnofsky quality of life scores compared to group B after six months (P=0.027). The Enneking scores of limb functions in group A were significantly higher than those of group B (P=0.022). In group A the postoperative limb function score was good and excellent in 92% and in group B in 64.7%. There were significantly more postoperative complications in group B compared with group A (P=0.027), but no significant difference in the recurrence rate in the two group (P=0.976). The study results can provide reference for surgical treatment of the patients with malignant bone tumors.

Effect of branched chain amino acids on perioperative temperature, glucose level and fat metabolism in patients with gastrointestinal tumors.

This study aimed to investigate the effect of branched chain amino acids (BCCAs) on perioperative temperature, glucose and fat metabolism in patients with gastrointestinal tumors. Fifty-six patients undergoing gastrointestinal tumor surgery were included in the study and randomly divided into two groups of 28 patients each: an experimental and a control group. During surgery, the experimental group received 5.64mL·Kg-1·h-1(4KJ·Kg-1·h-1) of BCCAs intravenously, through an infusion pump, and the control group received an equal volume of NaCl 0.9%. Vital signs were continuously monitored during the operation. Nasopharynx temperature levels of glucose, insulin, free fatty acid and ketone bodies in the blood were determined 30 min before anesthesia (t 0), after anesthesia and before surgery (t 1), 30 min after the start of surgery (t 2), 2 h after start of surgery (t 3) and 1 h after the end of surgery (T4). Patients’ shivering intensity (Wrench grading) and pain degree [Visual analogue scale (VAS)]) were estimated 1 h after the endotracheal tube was removed. Nasopharynx temperature was decreased (p less than 0.05) in both groups after anesthesia induction, while 1 h after the tube was removed it was higher in the experimental group than the control group (p less than 0.05); compared with pre-surgery values, blood glucose levels were increased during surgery in both groups, but the experimental group had a lower increasing trend compared to the control group, though without statistical significance (p>0.05). Insulin levels were significantly different between the two groups at all time-points during surgery (p less than 0.05). However, the rising trend of the experimental group was more dramatic during the period from t 0 to t 3. One hour after surgery (t 4), the insulin levels varied, but still at higher levels than pre-surgery, with a significant difference (p less than 0.05); levels of free fatty acids had a downward trend in both groups, and levels in the experimental group continued to decline until 1 h after surgery. Patients who received branched chain amino acids had less temperature decrease during surgery. Moreover, blood glucose levels were not increased, which limits fat mobilization and leads to production of ketone bodies, reduces the shivering and its intensity after surgery.

Interleukin-2 reverses CD8(+) T cell exhaustion in clinical malignant pleural effusion of lung cancer.

Malignant pleural effusion (MPE) is a poor prognostic sign for cancer patients, whereas the functional condition of MPE CD8(+) T cells is unknown. Intracavitary immunotherapy with interleukin (IL)-2 has been proven effective in controlling MPE. To elucidate the underlying mechanism, 35 lung cancer (LC) patients with MPE and 12 healthy donors were included in this study. For the IL-2 therapy experiments, after draining partial MPE, we treated 14 patients by administrating IL-2 (3 or 5 × 10(6) U in 50 ml saline) into the thoracic cavity. Before and after IL-2 treatment (40-48 h), the MPE and peripheral blood (PB) were obtained from the subjects. PB from healthy volunteers was collected as control. The expression of programmed cell death 1 (PD-1), granzyme B (GzmB), interferon (IFN)-γ and the proliferation were analysed in CD8(+) T cells from MPE and PB. The CD8(+) T cells in the MPE of LC patients showed lowest GzmB, IFN-γ and proliferation but highest PD-1 expression, compared with that in PB of LC patients and healthy donors. IL-2 treatment reduced the expression of PD-1, increased the expression of GzmB and IFN-γ and enhanced the proliferation of CD8(+) T cells in MPE. In addition, IL-2 treatment reduced carcino-embryonic antigen (CEA) level in MPE. These results indicate that MPE CD8(+) T cells exhibit exhaustion phenotype which can be reversed by IL-2 therapy.

Impact of multimodal therapy in locally recurrent rectal cancer.

BACKGROUND: The practice of salvaging recurrent rectal cancer has evolved. The aim of this study was to define the evolving salvage potential over time among patients with locally recurrent disease, and to identify durable determinants of long-term success. METHODS: The study included consecutive patients with recurrent rectal cancer undergoing multimodal salvage with curative intent between 1988 and 2012. Predictors of long-term survival were defined by Cox regression analysis and compared over time. Re-recurrence and subsequent treatments were evaluated. RESULTS: After multidisciplinary evaluation of 229 patients, salvage therapy with curative intent included preoperative chemotherapy and/or radiotherapy (73·4 per cent; with 41·3 per cent undergoing repeat pelvic irradiation), surgical salvage resection with or without intraoperative irradiation (36·2 per cent), followed by postoperative adjuvant chemotherapy (38·0 per cent). Multivisceral resection was undertaken in 47·2 per cent and bone resection in 29·7 per cent. The R0 resection rate was 80·3 per cent. After a median follow-up of 56·5 months, the 5-year overall survival rate was 50 per cent in 2005-2012, markedly increased from 32 per cent in 1988-1996 (P = 0·044). Long-term success was associated with R0 resection (P = 0·017) and lack of secondary failure (P = 0·003). Some 125 patients (54·6 per cent) developed further recurrence at a median of 19·4 months after salvage surgery. Repeat operative rescue was feasible in 21 of 48 patients with local re-recurrence alone and in 17 of 77 with distant re-recurrence, with a median survival of 19·8 months after further recurrence. CONCLUSION: The long-term salvage potential for recurrent rectal cancer improved significantly over time, with the introduction of an individualized treatment algorithm of multimodal treatments and surgical salvage. Durable predictors of long-term success were R0 resection at salvage operation, avoidance of secondary failure, and feasibility of repeat rescue after re-recurrence.

Genetic variations of CD36 and low platelet CD36 expression - a risk factor for lipemic plasma donation in Taiwanese apheresis donors.

BACKGROUND: New CD36 mutations are constantly being identified, although no study has specifically targeted a Taiwanese population. CD36 deficiency can result in dyslipid state and slow clearance of chylomicron. This could be linked to more frequent lipemic donations. STUDY DESIGN AND METHODS: We used flow cytometric methods to study the CD36 deficiency in 640 regular volunteer platelet apheresis donors from Taipei blood centre. The coding exons of CD36 gene were sequenced in CD36-deficient individuals, and the allele frequencies of CD36 variants were determined in the larger population by mutation-specific PCR and oligonucleotide hybridization. Visual inspection of lipemic plasma was routinely performed on samples taken before commencement of apheresis. Individuals found to have lipemic plasma are deferred until next donation. We investigated the link between positive lipemic deferral record and low platelet CD36 expression status. RESULTS: We found four donors (0·6%) with type I CD36 deficiency (both platelets and monocytes CD36(null) ) and six (1·0%) with type II CD36 deficiency (PLT: CD36(null) , monocyte: CD36(low) ). Six CD36 genetic variants were identified, two of them were novel, all but one are found exclusively in CD36(null) and CD36(low) expressors. Subjects with CD36 genetic variants also displayed deficient or reduced CD36 on monocytes. Donors with null or low PLT CD36 expression were more likely to have a lipemic deferral record than control subjects with normal PLT CD36 expression (X(2) = 27·36, odds ratio = 2·6, 95% conference interval: 1·8-3·8, P < 0·0001). CONCLUSION: Through this study, we established a donor registry to supply CD36-negative platelets for patients in need.

Epidemiological and clinical characteristics of severe fever with thrombocytopenia syndrome (SFTS) in China: an integrated data analysis.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease that was caused by a novel bunyavirus, SFTSV. The study aimed to disclose the epidemiological and clinical characteristics of SFTSV infection in China so far. An integrated clinical database comprising 1920 SFTS patients was constructed by combining first-hand clinical information collected from SFTS sentinel hospitals (n = 1159) and extracted data (n = 761) from published literature. The considered variables comprised clinical manifestations, routine laboratory tests of acute infection, hospitalization duration and disease outcome. SFTSV-IgG data from 19 119 healthy subjects were extracted from the published papers. The key clinical variables, case-fatality rate (CFR) and seroprevalence were estimated by meta-analysis. The most commonly seen clinical manifestations of SFTSV infection were fever, anorexia, myalgia, chill and lymphadenopathy. The major laboratory findings were elevated lactate dehydrogenase, aminotransferase, followed by thrombocytopenia, lymphocytopenia, elevated alanine transaminase and creatine kinase. A CFR of 12·2% was estimated, significantly higher than that obtained from national reporting data, but showing no geographical difference. In our paper, the mortality rate was about 1·9 parts per million. Older age and longer delay to hospitalization were significantly associated with fatal outcome. A pooled seroprevalence of 3·0% was obtained, which increased with age, while comparable for gender. This study represents a clinical characterization on the largest group of SFTS patients up to now. A higher than expected CFR was obtained. A wider spectrum of clinical index was suggested to be used to identify SFTSV infection, while the useful predictor for fatal outcome was found to be restricted.

Polymorphism of X-linked CD40 ligand gene associated with pulmonary tuberculosis in the Han Chinese population.

Among those developing tuberculosis (TB) after exposure to Mycobacterium tuberculosis, approximately 70% are males. Host genetic variation, particularly immune-related genes on the X chromosome, may contribute to sex-specific differences in TB incidences. To study whether X-linked gene variation is associated with sex-specific presentation of pulmonary TB (pTB), three single-nucleotide polymorphisms (SNPs) of the TLR8, CD40LG and IRAK1 genes on the X chromosome were genotyped in 923 patients and 1033 healthy individuals of the Han Chinese population. Frequencies of the variants were analyzed independently as well as in their combinations. CD40LG rs3092923 and its combined effects with the other two SNPs were associated with an increased risk of pTB only in males. In males, the rs3092923 genotype C/(-) conferred relative protection (odds ratio (OR): 0.52, 95% confidence interval (CI): 0.35-0.78, Pcorr.=0.0045) and the combined effects of three SNPs increased gradually as the number of risk alleles increased (OR: 2.58, 2.83 and 2.96 for one, two and three risk alleles, respectively). For the remaining SNPs, significance was obtained only for the AA genotype of IRAK1 rs3027898 in the combined and female-only analysis. Our results indicate a role of a CD40LG variant and its combined effects with distinct TLR8 and IRAK1 variants in susceptibility to pTB in males.

Interaction of thymic stromal lymphopoietin, IL-33, and their receptors in epithelial cells in eosinophilic chronic rhinosinusitis with nasal polyps.

BACKGROUND: Thymic stromal lymphopoietin (TSLP), IL-25, and IL-33 system contribute to the initiation and development of Th2 responses. This study aimed to explore the involvement of TSLP, IL-25, IL-33, and their receptors in type 2 T-helper (Th) responses in chronic rhinosinusitis with nasal polyps (CRSwNPs) and their cross-regulation in human nasal epithelial cells (HNECs). METHODS: Immunohistochemistry, quantitative RT-PCR, ELISA, Bio-Plex assay, and flow cytometry were used to detect the expression of TSLP/common γ-like TSLP receptor (TSLPR)/IL-7 receptor α (IL-7Rα), IL-25/IL-17B receptor (IL-17RB), and IL-33/membrane-bound ST2 (ST2L)/soluble ST2 (sST2) in sinonasal mucosa and HNECs. HNECs cultured at an air-liquid interface were used to explore the expression in regulation of these cytokine systems. RESULTS: Compared with controls and noneosinophilic CRSwNP, the expression of TSLP/TSLPR/IL-7Rα and ST2L/sST2 was significantly increased in eosinophilic CRSwNP, predominantly in epithelial cells. In contrast, the expression of IL-33 and IL-25/IL-17RB was enhanced in epithelial cells in both eosinophilic and noneosinophilic CRSwNP compared to controls. The expression of TSLP, TSLPR, and ST2L was positively correlated with symptom and computer tomography scan scores in eosinophilic CRSwNP and with Th2 cytokine expression in sinonasal mucosa. The expression of ST2L was correlated with TSLP and its receptor expression. TSLP could induce ST2L expression that promoted IL-33-induced TSLP expression in HNECs. In addition, TSLP/TSLPR/IL-7Rα and ST2L could be induced by Th2 cytokines, while IL-25/IL-17RB and IL-33 could be upregulated by Th1/Th17 cytokines, in HNECs. CONCLUSIONS: The positive feedback loop between TSLP, IL-33 and their receptors, and Th2 cytokines may facilitate Th2-skewed inflammation in eosinophilic CRSwNP.

Development of primer pairs from diverse chloroplast genomes for use in plant phylogenetic research.

Variation in the chloroplast DNA sequence is useful for plant phylogenetic studies. However, the number of variable sequences provided by chloroplast DNA for suggested genes or genomic regions in plant phylogenetic analyses is often inadequate. To identify conserved regions that can be used to design primers and amplify variable sequences for use in plant phylogenetic studies, the complete chloroplast genomic sequences of six plant species (including Oryza sativa, Arabidopsis thaliana, Glycine max, Lotus japonicus, Medicago truncatula, and Phaseolus vulgaris), searched from the taxonomy database of NCBI were investigated. A total of 93 conserved regions, 32 in large single copy and 61 in inverted repeat regions, were identified. A set of five primer pairs were designed according to the conserved sequences located in the psbA~trnK, psbB~psbH, rpl23~trnI, trnR~trnN, and trnY~trnD regions to amplify variable DNA fragments. An additional 18 plant accessions from 14 species were used to validate their utility. Each of the tested species could be distinguished by length polymorphisms of fragments amplified with the five primer pairs. trnR~trnN and rpl23~trnI amplified fragments specific to monocot and legume species, respectively. Three primer pairs located in the psbA~trnK, psbB~psbH, and trnR~trnN regions were applied to amplify variable DNA sequences for phylogenetic analysis using the maximum parsimony method. The consistent result between taxonomy and phylogenetic analysis on the variable sequences amplified with these three primer pairs was revealed. The five newly developed primer pairs are recommended as tools for use in the identification of plant species and in phylogenetic studies.