BACKGROUND: Double-stranded RNA-specific adenosine deaminase 1 (ADAR1) binds to double-stranded RNA and catalyzes the deamination of adenosine (A) to inosine (I). The functional mechanism of ADAR1 in non-small cell lung cancer (NSCLC) remains incompletely understood. This study aimed to investigate the prognostic significance of ADAR1 in NSCLC and to elucidate its potential role in regulating tumor cell proliferation and migration. METHODS: Data from The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed to assess the correlation between high ADAR1 expression and clinicopathological features as well as prognosis in lung cancer. We performed Western blot (WB), cell proliferation assays, Transwell invasion/migration assays, and nude mouse xenograft modeling to examine the phenotypic changes and molecular mechanisms induced by ADAR1 knockdown. Furthermore, the ADAR1 p150 overexpression model was utilized to validate the proposed mechanism. RESULTS: ADAR1 expression was significantly elevated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues compared with adjacent non-tumor tissues (LUAD: P=3.70×10-15, LUSC: P=0.016). High ADAR1 expression was associated with poor prognosis (LUAD: P=2.03×10-2, LUSC: P=2.81×10-2) and distant metastasis (P=0.003). Gene Set Enrichment Analysis (GSEA) indicated that elevated ADAR1 was associated with mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway activation, matrix metalloproteinase-9 (MMP-9) expression, and cell adhesion. ADAR1 and MMP-9 levels showed a strongly positive correlation (P=6.45×10-34) in 10 lung cancer cell lines, highest in H1581. Knockdown of ADAR1 in H1581 cells induced a rounded cellular morphology with reduced pseudopodia. Concomitantly, it suppressed cell proliferation, invasion, migration, and in vivo tumorigenesis. It also suppressed ERK phosphorylation and downregulated cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-FOS), MMP-9, N-cadherin, and Vimentin. Conversely, ADAR1 p150 overexpression in PC9 cells enhanced ERK phosphorylation and increased c-FOS and MMP-9 expression. CONCLUSIONS High ADAR1 expression is closely associated with poor prognosis and distant metastasis in NSCLC patients. Mechanistically, ADAR1 may promote proliferation, invasion, migration, and tumorigenesis in lung cancer cells via the ERK/c-FOS/MMP-9 axis.
Humans ; Lung Neoplasms/physiopathology* ; Adenosine Deaminase/genetics* ; Matrix Metalloproteinase 9/genetics* ; Cell Proliferation ; Carcinoma, Non-Small-Cell Lung/physiopathology* ; Cell Movement ; Animals ; Mice ; RNA-Binding Proteins/genetics* ; Female ; Male ; Cell Line, Tumor ; Proto-Oncogene Proteins c-fos/genetics* ; Middle Aged ; MAP Kinase Signaling System ; Gene Expression Regulation, Neoplastic ; Mice, Nude ; Extracellular Signal-Regulated MAP Kinases/genetics*

BACKGROUND: The switch/sucrose nonfermentable chromatin-remodeling (SWI/SNF) complex is a pivotal chromatin remodeling complex, and the genomic alterations (GAs) of the SWI/SNF complex are observed in several cancer types, correlating with multiple biological features of tumor cells. However, their role in liver metastasis of non-small cell lung cancer (NSCLC) remains unclear. Our study aims to investigate the role and potential mechanisms underlying NSCLC liver metastasis induced by the GAs of SWI/SNF complex. METHODS: The GAs of SWI/SNF complex in NSCLC cell lines (H1299, H23 and H460) were identified by whole-exome sequencing (WES). ARID1A knockout H1299 cell was constructed with the CRISPR/Cas9 technology. The mouse model of liver metastasis from NSCLC was established to simulate lung cancer liver metastasis and observe the metastasis rate under different gene mutation conditions. RNA sequencing and Western blot were conducted for differential gene expression analysis. Immunohistochemistry (IHC) analysis was used to assess protein expression levels of SWI/SNF-regulated target molecules in mouse liver metastases. RESULTS: WES analysis revealed intracellular gene mutations. The animal experiments demonstrated a correlation between the GAs of SWI/SNF complex and a higher liver metastasis rate in immunodeficient mice. Transcriptome sequencing and Western blot analysis showed upregulated expression of ALDH1A1 and APOBEC3B in SWI/SNF-mut cells, particularly in ARID1A-deficient H460 and H1299 sgARID1A cells. IHC staining of mouse liver metastases further demonstrated elevated expression of ALDH1A1 in the H460 and H1299 sgARID1A group. CONCLUSIONS This study underscores the critical role of the GAs of SWI/SNF complex, such as ARID1A and SMARCA4, in promoting liver metastasis of lung cancer cells. The GAs of SWI/SNF complex may promote liver-specific metastasis by upregulating ALDH1A1 and APOBEC3B expression, providing novel insights into the molecular mechanisms underlying lung cancer liver metastasis.
Animals ; Mice ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Mutation ; Liver Neoplasms/genetics*

【Objective】 To establish a method for batch weighing blood components intelligently and accurately. 【Methods】 Virus inactivated plasma was used as weighing samples. The traditional electric scale and cryoprecipitation preparation instrument were used to weigh plasma with five specifications. The consistency of the weighing results by the two methods was statistically analyzed. Two groups of virus inactivated plasma with basically the same number of bags were randomly selected, and weighted by the same group of staff using these two equipment, the time for weighing and the time from weighing to registering were calculated statistically. 【Results】 227 bags of plasma were weighed in batches by the two methods, and statistical analysis showed no significant difference in standardized blood volume. There was no significant difference in plasma weighing time (71/68)by the same staff, weighing basically the same number of bags. However, the time difference from weighing to registering was statistically significant, as the new method was 2.65 s shorter than the traditional one (P<0.05). 【Conclusion】 The new method is accurate for batch weighing, and can realize the informatization and intellectualization of blood weighting. The time consumed from weighing to registration was improved by the new method, but not in time for weighing.

Objective:To explore the incidence, clinical characteristics and prognosis of invasive pulmonary fungal infection(IPFI)in recipients of lung transplantation(LT)in southern China.Methods:From January 2003 to August 2019, retrospective analysis was performed for 300 recipients of lung transplantation at three hospitals in southern China. There were 254 males and 46 females with an average age of (54.98±14.2)years. Clinical data were collected from medical records, including symptoms and signs, imaging studies, bronchoscopy examination, pathogen separation and culture from deep sputum and bronchoalveolar lavage fluid(BALF), fungal-related laboratory tests and tissue pathology.Results:Among 300 cases, 93(31.0%)had at least one episode of IPFI. The most common pathogen was aspergillosis(60.2%), followed by candida(15 cases, 16.1%)and Pneumocystis jeroveci (13 cases, 14.0%). Kaplan Meier analysis indicated that all-cause mortality was significantly higher in IPFI group than that in non-IPFI(nIPFI)group with one-year mortality of 45.2% vs. 26.7% in IPFI and nIPFI groups respectively( P<0.05). Conclusions:IPFI is prevalent after LT in southern China. And aspergillosis is the most common pathogen and Candida comes the next. The median occurring time for aspergillosis is 6 months after LT. Candida infection occurs earlier at airway anastomosis. A higher incidence of invasive fungal disease(IFD)associated with a lower survival indicates that IPFI has a substantial mortality among recipients after LT. Prophylactic agents should be optimized based upon an epidemiologically likely pathogen.

Guideline or consensus for the vaccination of children with special conditions like immunocompromised children may be more suitable for pediatricians.However, the vaccination of children in China is mainly performed by general practitioners or child health care practitioners in community health service centers.They need to master the screening knowledge of contraindications and precautions for the vaccination of children, and make the decision to referral to specialists.Based on the technical guidelines for Immunization of National Health Commission of the People′s Republic of China, Best Practices Guidance of the Advisory Committee on Immunization Practices and contraindications and precautions proposed by the Immunization Action Coalition, 20 suggestions for primary screening of children with vaccination contraindications and referral recommendations for primary care providers were developed by experts from the Integration of Medicine and Prevention in Children of Health Commission of Shenzhen Municipality.

Objective:To explore the diagnosis and treatment strategies of primary ciliary dyskinesia (PCD).Methods:A 37-year-old male recipient who had repeated cough and sputum from childhood, with shortness of breath after activity and progressive development, the number of hospitalizations per year was ≥6 times, the MRC score was 2~3 diagnosed with primary ciliary dyskinesia through medical examination and multidisciplinary consultation, and received allogeneic double lung transplantation after medical treatment failure. Search related domestic and foreign literatures to explore and analyze the etiology, pathogenesis, clinical manifestations and imaging features, diagnosis and treatment of PCD.Results:PCD is an autosomal hereditary disease. Due to abnormal skeletal structure and/or functional development, clinical manifestations are mostly chronic sinusitis, bronchiectasis, otitis media and infertility. Chest imaging showed situs inversus totalis, right heart and diffuse cystic bronchiectasis infection with bronchial wall thickening, diagnosis depends on clinical manifestations, saccharin test, nasal nitric oxide test, high-speed video microscopy analysis, transmission electron microscopy, genetic testing and immunofluorescence.Conclusions:Lung transplantation is the only effective treatment for end-stage PCD. It not only saves patients' lives, but also significantly improves their quality of life.

OBJECTIVE: To explore the genetic basis for a child featuring X-linked intellectual disability. METHODS: The 1-year-and-6-month-old child presented with growth retardation, intellectual disability and bilateral alternating squint. With DNA extracted from the child and his parents' peripheral venous blood samples, whole exome sequencing was carried out to identify potential variants that can explain his condition. Suspected variants were validated by Sanger sequencing. The impact of variants was predicted by bioinformatic tools. RESULTS: The child was found to harbor a de novo nonsense c.3163C>T (p.Arg1055*) variant of the IQSEC2 gene. The variant, unreported previously, was predicted to be pathogenic based on MutationTaster, PROVEAN and SIFT. Analysis using a HomoloGene system suggested Arg1055 in IQSEC2 residues to be highly conserved evolutionarily, and that replacement of Arg1055 may cause destroy of the PH domain (AA 951-1085) and serious damage to the function of IQSEC2 protein. Analysis with UCSF chimera software suggested that the c.3163C>T (p.Arg1055*) variant can induce serious damages to the secondary structures of IQSEC2 protein, causing loss of its function. CONCLUSION The patient's condition may be attributed to the de novo nonsense variant c.3163C>T (p.Arg1055*) of the IQSEC2 gene.

Objective: To detect pathogenic variant of ARSA gene in an infant with late infantile metachromatic leukodystrophy (MLD). Methods: The male proband had an onset of walking dysfunction and seizure at 28 months. Arylsulfatase A activity of his peripheral blood leucocytes was 26.9 nmol/mg.17h, and cranial MRI showed wild symmetrical demyelination. With genomic DNA extracted from his peripheral blood sample, all coding exons and splicing sites of the ARSA gene were subjected to Sanger sequencing. PubMed Protein BLAST system was employed to analyze cross-species conservation of the mutant amino acid. Ucsf chimera software was used to analyze the impact of candidate variants on the secondary structure of the protein product. Impact of potential variants was also analyzed with software including PolyPhen-2, Mutation Taster, SIFT and PROVEAN. Whole-exome sequencing was carried out to identify additional variants which may explain the patient’s condition. Results: The proband was found to harbor compound heterozygous variants of the ARSA gene [c.467G>A (p.Gly156Asp) and c. 960G>A (p.Trp320*)], neither of which was reported previously. As predicted by Ucsf chimera software, the c. 960G>A (p.Trp320*) variant may demolish important secondary structures including α-helix, β-strand and coil of the ARSA protein, causing serious damage to its structure and loss of function. The c. 467G>A (p.Gly156Asp) variant was predicted to be "probably damaging" by PolyPhen-2, Mutation Taster and SIFT software. Conclusion The patient’s condition may be attributed to the compound heterozygous c. 467G>A (p.Gly156Asp) and c. 960G>A (p.Trp320*) variants of the ARSA gene. Above results have facilitated genetic counseling and prenatal diagnosis for this family.

Objective:To evaluate the effect of reactivated ultrasound contrast agent on prevention of ischemia-reperfusion (IR) injury in rat hindlimb ischemia model.Methods:Microbubbles were compressed into nanodroplets (NDs) and the particle size range was determined.In vitro experiments were carried out to observe the acoustic phase transition of nanodroplets in the ultrasound field and the change of dissolved oxygen (DO) concentration in the surrounding solution. Forty-one male Sprague Dawley rats were divided into 5 groups: acoustic nanodroplet vaporization treatment group (NDs+ US group, n=9), saline+ ultrasound treatment group (Saline+ US group, n=8), nanodroplet without ultrasound treatment group (NDs group, n=8), ischemia/reperfusion injury group (IRI group, n=8) and sham operation group (Sham group, n=8). Ultrasound imaging was performed before operation and 12 hours after reperfusion to evaluate the improvement of hemodynamics of criminal artery under different treatments. Tissue injury were evaluated by analyzing immunohistochemistry staining results. Results:The formed NDs ranged in size from approximately 68.0-295.4 nm and the highest concentration in the 100 nm range. In vitro studies, a decrease in DO was measured during the phase transition.In the animal experiment, after ischemia/reperfusion, NDs+ US, Saline+ US, NDs and IRI groups demonstrated a significantly higher resistance index (RI) and pualsatility index (PI) of the right common iliac artery compared with before operation (NDs+ US group: PI 1.79±0.17 vs 1.57±0.23, P=0.014; RI 0.80±0.02 vs 0.75±0.04, P=0.002. Saline+ US group: PI 2.29±0.16 vs 1.57±0.16, P<0.001; RI 0.90±0.06 vs 0.74±0.03, P<0.001. NDs group: PI 2.17±0.14 vs 1.53±0.15, P<0.001; RI 0.91±0.04 vs 0.75±0.04, P<0.001. IRI group: PI 2.12±0.22 vs 1.58±0.20, P<0.001; RI 0.88±0.04 vs 0.75±0.04, P<0.001). The increases of PI and RI (ΔPI, ΔRI) in NDs+ US group were higher than those in sham group (all P<0.05), but significantly lower than those in saline+ US group, NDS group and IRI group (all P<0.05). Immunohistochemical results indicated the percentages malondialdehyde (MDA) positive cells in Saline+ US, NDs and IRI groups were higher than those in NDs+ US and sham groups (all P<0.05). Conclusions:The acoustic phase transition of nanodroplets in the ultrasound field can reduce ischemia/reperfusion injury and improve hemodynamics abnormality after reperfusion.

In recent years, more and more reports have been reported on the treatment of refractory primary membranous nephropathy with rituximab in bed, and its effect has been confirmed.A case of refractory primary membranous nephropathy was treated in the Department of Nephrology, Peking university Third Hospital.After the treatment of hormone combined with cyclophosphamide, cyclosporine and tacrolimus was ineffective, the patients were treated with low-dose rituximab for 2 years according to their condition, and the condition of the patients was finally relieved.