[New 2023 global definition of acute respiratory distress syndrome: progress and limitation].

Acute respiratory distress syndrome (ARDS) presents a challenge in clinical diagnosis as it lacks a definitive gold standard. Over the past 55 years, there have been several revisions to the definition of ARDS. With the progress of clinical practice and scientific research, the limitations of the "Berlin definition" have become increasingly evident. In response to these changes, the 2023 global definition of ARDS aims to address these issues by expanding the diagnostic targets, chest imaging, and methods for assessing hypoxia. Additionally, the new definition increases the diagnostic criteria to accommodate resource-constrained settings. The expansion facilitates early identification and treatment interventions for ARDS, thereby advancing epidemiological and clinically related research. Nevertheless, the broad nature of this revision may include patients who do not actually have ARDS, thus raising the risk of false-positive diagnoses. Therefore, additional verification is crucial to ascertain the validity and accuracy of the 2023 global definition of ARDS.

[Effects of microRNA-106b on migration and invasion of human malignant pleural mesothelioma cell NCI-H2452].

Objective: To investigate the inhibitory effect of microRNA-106b in the process of migration and invasion of human malignant pleural mesothelioma cell NCI-H2452. Methods: In April 2017, the expression level of miRNA-106b in malignant pleural mesothelioma cells (NCI-H2452, MSTO-211H, NCI-H2052) and normal mesothelial cells MeT-5A was detected and analyzed. Using NCI-H2452 cells as a model, the NCI-H2452 cell model with miRNA-106b overexpression was established by transfecting miRNA-106b mimics. The expression level of miRNA-106b in the cells was detected by real-time fluorescent quantitative PCR. The effect of miRNA-106b on the migration and invasion ability of NCI-H2452 cells was analyzed. The gene expression data of malignant mesothelioma and the downstream target gene data of miRNA-106b in public databases were analyzed to screen the downstream target genes of miRNA-106b in mesothelioma cells that affect cell migration and invasion ability, and to verify the expression of this gene in NCI-H2452 cells with miRNA-106b overexpression. Results: The expression of miRNA-106b in three MPM cells was decreased compared with MeT-5A cells (P<0.001) . The expression level of miRNA-106b was significantly increased after transfection of miRNA-106b mimics (P<0.001) . The scratch migration levels of the experimental group were 28.45%±4.37%, 38.12%±4.82% and 50.06%±8.92% at 24h, 31h and 48h, respectively. Compared with the control group, the migration level decreased by 37.48%±2.65%, 49.21%±3.45% and 68.14%±3.81% (P<0.01) . The number of cell migration and invasion decreased in the experimental group compared with the control group (P<0.001) . Public databases were used to screen and analyze the possibility that TCF21 gene, as a downstream target gene, could affect the migration and invasion ability of MPM cells. The expression level of TCF21 gene was increased after transfection of miRNA-106b mimics in NCI-H2452 cells (P=0.009) . Conclusion: MiRNA-106b can inhibit the migration and invasion of NCI-H2452 cells and increase the expression of TCF21 gene.

[Evaluation and clinical significance of gas exchange in patients with acute respiratory distress syndrome].

Gas exchange abnormalities is the pathophysiology characteristic of acute respiratory distress syndrome (ARDS). The severity of gas exchange abnormalities not only reflect the severity and outcome of the disease, but could also be an important index to guide individual mechanical ventilation settings and evaluate the therapeutic effects of inhaled vasodilator. The common techniques to measure gas exchange include multiple inert gas elimination technique, automatic lung parameter estimator, electrical impedance tomography, and single-photon emission CT. Nowadays, bedside techniques and measurements for improving gas exchange function in ARDS patients are still limited. Therefore, the improvement and promotion of bedside real-time gas exchange monitoring technology may achieve the goal of personalized medicine in ARDS. This article reviewed the common evaluation methods of gas exchange function in ARDS and their significance, in order to pay more attention to the evaluation of gas exchange function and further improve the prognosis of patients with ARDS.

[Effects of in situ cross-linked graphene oxide-containing gelatin methacrylate anhydride hydrogel on wound vascularization of full-thickness skin defect in mice].

Objective: To prepare graphene oxide (GO)-containing gelatin methacrylate anhydride (GelMA) hydrogel and to investigate the effects of in situ photopolymerized GO-GelMA composite hydrogel in wound vascularization of full-thickness skin defect in mice. Methods: The experimental study method was used. The 50 µL of 0.2 mg/mL GO solution was evenly applied onto the conductive gel, and the structure and size of GO were observed under field emission scanning electron microscope after drying. Human skin fibroblasts (HSFs) were divided into 0 µg/mL GO (without GO solution, the same as below) group, 0.1 µg/mL GO group, 1.0 µg/mL GO group, 5.0 µg/mL GO group, and 10.0 µg/mL GO group treated with GO of the corresponding final mass concentration, and the absorbance value was detected using a microplate analyzer after 48 h of culture to reflect the proliferation activity of cells (n=6). HSFs and human umbilical vein vascular endothelial cells (HUVECs) were divided into 0 µg/mL GO group, 0.1 µg/mL GO group, 1.0 µg/mL GO group, and 5.0 µg/mL GO group treated with GO of the corresponding final mass concentration, and the migration rates of HSFs at 24 and 36 h after scratching (n=5) and HUVECs at 12 h after scratching (n=3) were detected by scratch test, and the level of vascular endothelial growth factor (VEGF) secreted by HSFs after 4, 6, and 8 h of culture was detected by enzyme-linked immunosorbent assay method (n=3). The prepared GO-GelMA composite hydrogels containing GO of the corresponding final mass concentration were set as 0 µg/mL GO composite hydrogel group, 0.1 µg/mL GO composite hydrogel group, 1.0 µg/mL GO composite hydrogel group, and 5.0 µg/mL GO composite hydrogel group to observe their properties before and after cross-linking, and to detect the release of GO after soaking with phosphate buffer solution for 3 and 7 d (n=3). The full-thickness skin defect wounds were made on the back of 16 6-week-old female C57BL/6 mice. The mice treated with in situ cross-linked GO-GelMA composite hydrogel containing GO of the corresponding final mass concentration were divided into 0 µg/mL GO composite hydrogel group, 0.1 µg/mL GO composite hydrogel group, 1.0 µg/mL GO composite hydrogel group, and 5.0 µg/mL GO composite hydrogel group according to the random number table, with 4 mice in each group. The general condition of wound was observed and the wound healing rate was calculated on 3, 7, and 14 d of treatment, the wound blood perfusion was detected by laser Doppler flowmetry on 3, 7, and 14 d of treatment and the mean perfusion unit (MPU) ratio was calculated, and the wound vascularization on 7 d of treatment was observed after hematoxylin-eosin staining and the vascular density was calculated (n=3). The wound tissue of mice in 0 µg/mL GO composite hydrogel group and 0.1 µg/mL GO composite hydrogel group on 7 d of treatment was collected to observe the relationship between the distribution of GO and neovascularization by hematoxylin-eosin staining (n=3) and the expression of VEGF by immunohistochemical staining. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey's method. Results: GO had a multilayered lamellar structure with the width of about 20 µm and the length of about 50 µm. The absorbance value of HSFs in 10.0 µg/mL GO group was significantly lower than that in 0 µg/mL GO group after 48 h of culture (q=7.64, P<0.01). At 24 h after scratching, the migration rates of HSFs were similar in the four groups (P>0.05); at 36 h after scratching, the migration rate of HSFs in 0.1 µg/mL GO group was significantly higher than that in 0 µg/mL GO group, 1.0 µg/mL GO group, and 5.0 µg/mL GO group (with q values of 7.48, 10.81, and 10.20, respectively, P<0.01). At 12 h after scratching, the migration rate of HUVECs in 0.1 µg/mL GO group was significantly higher than that in 0 µg/mL GO group, 1.0 µg/mL GO group, and 5.0 µg/mL GO group (with q values of 7.11, 8.99, and 14.92, respectively, P<0.01), and the migration rate of HUVECs in 5.0 µg/mL GO group was significantly lower than that in 0 µg/mL GO group and 1.0 µg/mL GO group (with q values of 7.81 and 5.33, respectively, P<0.05 or P<0.01 ). At 4 and 6 h of culture, the VEGF expressions of HSFs in the four groups were similar (P>0.05); at 8 h of culture, the VEGF expression of HSFs in 0.1 µg/mL GO group was significantly higher than that in 0 µg/mL GO group and 5.0 µg/mL GO group (with q values of 4.75 and 4.48, respectively, P<0.05). The GO-GelMA composite hydrogels in the four groups were all red liquid before cross-linking, which turned to light yellow gel after cross-linking, with no significant difference in fluidity. The GO in the GO-GelMA composite hydrogel of 0 µg/mL GO composite hydrogel group had no release of GO at all time points; the GO in the GO-GelMA composite hydrogels of the other 3 groups was partially released on 3 d of soaking, and all the GO was released on 7 d of soaking. From 3 to 14 d of treatment, the wounds of mice in the 4 groups were covered with hydrogel dressings, kept moist, and gradually healed. On 3, 7, and 14 d of treatment, the wound healing rates of mice in the four groups were similar (P>0.05). On 3 d of treatment, the MPU ratio of wound of mice in 0.1 µg/mL GO composite hydrogel group was significantly higher than that in 0 µg/mL GO composite hydrogel group, 1.0 µg/mL GO composite hydrogel group, and 5.0 µg/mL GO composite hydrogel group (with q values of 10.70, 11.83, and 10.65, respectively, P<0.05 or P<0.01). On 7 and 14 d of treatment, the MPU ratios of wound of mice in the four groups were similar (P>0.05). The MPU ratio of wound of mice in 0.1 µg/mL GO composite hydrogel group on 7 d of treatment was significantly lower than that on 3 d of treatment (q=14.38, P<0.05), and that on 14 d of treatment was significantly lower than that on 7 d of treatment (q=27.78, P<0.01). On 7 d of treatment, the neovascular density of wound of mice on 7 d of treatment was 120.7±4.1 per 200 times of visual field, which was significantly higher than 61.7±1.3, 77.7±10.2, and 99.0±7.9 per 200 times of visual field in 0 µg/mL GO composite hydrogel group, 1.0 µg/mL GO composite hydrogel group, and 5.0 µg/mL GO composite hydrogel group (with q values of 12.88, 7.79, and 6.70, respectively, P<0.01), and the neovascular density of wound of mice in 1.0 µg/mL GO composite hydrogel group and 5.0 µg/mL GO composite hydrogel group was significantly higher than that in 0 µg/mL GO composite hydrogel group (with q values of 5.10 and 6.19, respectively, P<0.05). On 7 d of treatment, cluster of new blood vessels in wound of mice in 0.1 µg/mL GO composite hydrogel group was significantly more than that in 0 µg/mL GO composite hydrogel group, and the new blood vessels were clustered near the GO; a large amount of VEGF was expressed in wound of mice in 0.1 µg/mL GO composite hydrogel group in the distribution area of GO and new blood vessels. Conclusions: GO with mass concentration lower than 10.0 µg/mL had no adverse effect on proliferation activity of HSFs, and GO of 0.1 µg/mL can promote the migration of HSFs and HUVECs, and can promote the secretion of VEGF in HSFs. In situ photopolymerized of GO-GelMA composite hydrogel dressing can promote the wound neovascularization of full-thickness skin defect in mice and increase wound blood perfusion in the early stage, with GO showing an enrichment effect on angiogenesis, and the mechanism may be related to the role of GO in promoting the secretion of VEGF by wound cells.

Characterizing the critical role of metabolism in osteosarcoma based on establishing novel molecular subtypes.

OBJECTIVE: Osteosarcoma is the third most frequently diagnosed cancer among adolescents. Immunotherapy is an effective curative treatment for metastatic osteosarcoma patients. This study aimed to further reveal the significance of metabolism in tumor progression, and to categorize molecular subtypes for guiding personalized therapy. MATERIALS AND METHODS: Univariate Cox regression analysis was performed to screen metabolism-related genes associated with osteosarcoma prognosis. A molecular subtyping system was developed by unsupervised consensus clustering. Survival analysis and functional analysis were used to evaluate the performance of subtyping and characterize the TME of subtypes. Stepwise Akaike information criterion (stepAIC) was employed to optimize the prognostic model. RESULTS: C1 and C2 subtypes showed distinct prognosis, with more favorable survival in C2 subtype. C2 subtype presented a higher immune infiltration and active anti-tumor response. Notably, C2 subtype was predicted to have better immune response to immune checkpoint blockade. In addition, a 5-gene prognostic signature with robust ability to classify patients into high-risk and low-risk groups was developed. CONCLUSIONS: The study revealed the critical role of metabolism in tumorigenesis by comparing the features between the two subtypes. Oncogenic pathways including epithelial mesenchymal transition (EMT), glycolysis and hypoxia may be closely involved in the correlation with metabolism. Importantly, we developed a novel subtyping system and a 5-gene signature with high potential to be applied in clinical practice.

[Analysis of upper airway structure development in children with different characteristics].

Objective: To analyze the development of upper airway in children with different characteristics. Methods: From June 2018 to June 2020, a total of 425 children younger than 16 years old who underwent head MRI examination and did not have sleep-disordered breathing were included in the study. The length of soft palate, tongue, upper airway, mental spine clivus, adenoid thickness and nasopharyngeal width were measured in the midsagittal plane of MRI image. Single factor variance analysis was used to compare the gender differences of upper airway parameters within certain age groups. Pearson correlation analysis was used to analyze the correlation between upper airway parameters and age. Results: The numbers of subjects in infant, young child, preschool, school age and adolescent group were 80, 86, 90, 90 and 79, respectively. There were 219 males, accounting for 51.5% of the study population. The adenoid thickness in the preschooler group was (1.26±0.26) cm, higher than that in the female group (1.15±0.20) cm (P=0.025). The upper airway length (5.89±0.60) cm and the ratio of upper airway length/mental spine-slope length (0.73±0.08) in males were higher than those in females [(5.31±0.45) cm and 0.67±0.07, respectively, P<0.05]. There was no gender difference in other upper airway parameters among different age groups (all P values>0.05). The length of upper airway, mental spine-slope, tongue, soft palate, the width of nasopharyngeal cavity and the thickness of adenoids were positively correlated with age (r=0.932, 0.912, 0.898, 0.705, 0.734 and 0.168, respectively), all P values<0.05. Adenoid thickness was positively correlated with age from birth to age 5 years (r=0.603, P<0.001), and negatively correlated with age after age 6 years (r=-0.259, P=0.001). Conclusion: There are gender differences in the development of upper airway structure in children of different ages.

[The impact of CD8 and CTLA-4 expression on histopathological character and survival in mesothelioma].

Objective: To investigate the survival and death risk factors of mesothelioma cases stratified by the expression levels of CD8 and cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) , providing new clue to evaluate disease progression and clinical outcome. Methods: This was a retrospective case report, which included 47 clinically and pathologically confirmed mesothelioma cases on November 2016. Their clinical and pathological information, asbestos exposure history and survival data were collected. Infiltrated lymphocyte, 5-methylcytosine (5-mC) , CTLA-4, CD8 and Ki-67 antigen were detected using hematoxylin-eosin staining and immunohistochemistry. Survival time and death risk factors of mesothelioma patients with different CD8 and CTLA-4 protein expression characteristics were analyzed. And analyze the influence of Ki-67 expression on the survival of patients with different CD8 and CTLA-4 protein and gene expression characteristics. Results: Among the 47 cases, 63.8% (30/47) had low/medium level of infiltrated lymphocyte. The immunohistochemistry scores of CTLA-4, CD8, 5-mC and Ki-67 were 92.97 (54.95, 120.65) , 72.41 (36.62, 89.82) , 11.09 (3.40, 52.89) and 5.88 (2.41, 11.48) , respectively. Patients with CD8(high) CTLA-4(high) had higher 5-mC level than those with CD8(high) CTLA-4(low) (P<0.01) . The median survival time of 27 cases was 0.83±0.29 year. The median survival times of those with CD8(high) CTLA-4(high) and CD8(high) CTLA-4(low) were 0.58±0.51 year and 0.83±0.30 year, respectively (P=0.521) . The immunohistochemistry score of Ki-67 ≥5.88 was an independent death risk factor for patients with CD8(high) CTLA-4(low) (HR=8.40, P=0.01) . Under different CD8 and CTLA-4 protein expression characteristics, in the patients with CD8(high) CTLA-4(low), the median survival times of those with high and low Ki-67 expression were 0.57±0.11 years and 2.31±0.46 years, respectively (P<0.01) . Under different CD8 and CTLA-4 mRNA expression characteristics, in the patients with CD8(high) CTLA-4(low), the median survival times of those with high and low Ki-67 mRNA expression were 1.20±0.36 years and 3.38±0.43 years, respectively (P=0.018) . Conclusion: Mesothelioma case with high CD8 but low CTLA-4 content might coexist DNA hypomethylation. In the presence of high Ki-67 expression, their survival time appears to be shortened with increased death risk.

[Expression and clinical significance of SETD2 in maligant pleural mesothelioma].

Objective: To analyze the gene mutation profile in malignant pleural mesothelioma (MPM) and investigate the expression of high-frequency mutant genes and its relationship with clinicopathological parameters. To screen out key genes and clinicopathologic factors related to the prognosis of MPM patients. Methods: The second generation sequencing data, somatic mutation data and clinical pathological data of 86 MPM cases and gene chip expression data of 89 MPM cases were downloaded from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) in March 2020. Summarize the gene mutation profile of tissue samples in the TCGA database and analyze the relationship between the expression level of high-frequency mutation genes and the clinicopathological characteristics, asbestos exposure history and prognosis of MPM patients. The genes significantly related to MPM prognosis were screened out for gene set enrichment analysis (GSEA) . Survival analysis and GSEA were performed for the selected key genes and clinicopathological features verification using the microarray expression data from the GEO database. Results: The top 10 genes with highest single nucleotide variations frequencies were BAP1, NF2, TP53, TTN, SETD2, LATS2, CCDC168, FAT4, PTCH1 and ZNF469. The high expression rates of NF2, TP53, SETD2 and CCDC168 genes in wild type were higher than those of mutated type, and the differences were statistically significant (P<0.05) . Cox multivariate analysis of TCGA data showed that MPM patients with epithelial type (HR=0.425, 95%CI: 0.235-0.767, P<0.01) and SETD2 low expression (HR=0.516, 95%CI: 0.307-0.868, P=0.011) had lower risk of death. The survival analysis of GEO data verified that patients with epithelial type MPM had longer survival time, while patients with sarcoma type MPM had shortest survival time (P<0.01) . GSEA showed that SETD2 was involved in G2M checkpoint, E2F targets, MYC signaling pathways, protein secretion, mitotic spindle, MTORC1 pathway, TGF-ß pathway, androgen response and uv response. Conclusion: MPM is accompanied with higher frequency of gene mutations represented by BAP1, NF2, TP53, TTN, SETD2, LATS2 and so on. SETD2 expression level and epithelia type of MPM may be influential factors for MPM prognosis.

[microRNA-30d can inhibit the proliferation, migration and invasion of human mesothelial cell MSTO-211H].

Objective: To investigate the inhibitory effect and molecular mechanism of microRNA-30d (miR-30d) in the process of proliferation, migration and invasion of malignant mesothelioma cell line MSTO-211H. Methods: In April 2017, the human MSTO-211H cells was used to establish miR-30d overexpressed MSTO-211H cell model by transfection of miR-30d mimics. The qRT-PCR was performed to detect the expression level of miR-30d in the cells transfected miR-30d mimics. The effects of miR-30d on the proliferation, apoptosis, migration and invasion of MSTO-211H cells were analyzed by CCK-8 experiment, flow cytometry, cell scratch experiment and Transwell method. Results: After transfection of miR-30d, the expression level of miR-30d in the MSTO-211H+miR-30d cells group was significantly higher than MSTO-211H+miR NC cells group (P<0.01) . The cell activity of MSTO-211H+miR-30d group (105.13%±2.35%) was significantly lower than MSTO-211H+miR NC cells group (115.40%±1.35%) , and the level of apoptosis (3.97%±0.36%) was significantly higher than MSTO-211H+miR NC cells group (1.47%±0.10%) (P<0.01) . The relative migration areas at 12 and 24 h of MSTO-211H+miR-30d cells group (9.35±3.16 µm(2) and 58.19±1.82 µm(2)) were significantly lower than MSTO-211H+miR NC cells group (54.42±5.26 µm(2) and 88.32±1.96 µm(2)) (P<0.01) . Compared with the MSTO-211H+miR NC cells group, the numbers of cell migration and cell invasion were reduced in the MSTO-211H+miR-30d cells group (P<0.01) . Conclusion: miR-30d can regulate the progression of malignant pleural mesothelioma by inhibiting the proliferation, apoptosis, migration and invasion of MSTO-211H cells.

[Study on malignant transformation of MeT-5A cells induced by chrysotile].

Objective: To study the cytotoxicity and malignant transformation ability of chrysotile on MeT-5A cells. Methods: In June 2016, lactate dehydrogenase (LDH) method was used to detect the cytotoxicity of chrysotile to MeT-5A cells. MeT-5A cells were treated with 5 µg/cm(2) chrysotile intermittently for 24 h a time, once a week and a total of 28 times. After the cells showed anchorage independent growth, the cell features of malignant transformation were identified by colony forming frequency in soft agar, and the soft agar colony formation rates were calculated. The activities of key speed limiting enzymes of glycolysis metabolism including hexokinase (HK) , phosphofructokinase (PFK) and pyruvate kinase (PK) were determined by UV colorimetry. Results: Chrysotile was cytotoxic to MeT-5A cells in a concentration-dependent decline. Compared with the control group, the relative survival rates of MeT-5A cells were significantly decreased after exposed to chrysotile at 10, 20, 40 and 80 µg/cm(2) (P<0.05) . After 28 times of exposure, the growth rate of the cells in chrysotile transformed MeT-5A cells was accelerated, the arrangement was disordered, the contact inhibition was lost, and the double layer growth appeared, which could grow on soft agar. The colony forming rate of the chrysotile transformed MeT-5A cells was 18.33‰±2.49‰. Compared with the control group (0) , the difference was statistically significant (P<0.01) . The activities of glycolysis related kinase including PK [ (19.51±1.52) U/L], PFK[ (0.12±0.02) U/10(4) cell] and HK[ (0.26±0.01) U/10(4) cell] were increased in the chrysotile transformed MeT-5A cells compared with control group [ (25.00±1.04) U/L、(0.15±0.01) U/10(4) cell and (0.33±0.01) U/10(4) cell] (P<0.01) . Conclusion: Chrysotile can induce malignant transformation of MeT-5A cells and increase the activities of glycolysis related kinases including PK, PFK and HK.

A combined approach using slightly acidic electrolyzed water and UV exposure to improve egg internal quality during storage.

This study investigated the combined efficacy of slightly acidic electrolyzed water (SAEW) and UV light (UV) in improving egg internal quality (weight loss, Haugh unit, yolk index, albumen pH) over a 6-wk storage time at 25°C. Eggs were preserved after immersion for 4 min in SAEW (30 mg/L), irradiation for 4 min under a UV lamp, or a combination of SAEW and UV treatment for 4 min. The combination of SAEW and UV inhibited the deterioration of yolk index over the storage period, as well as reducing the extent of decrease in Haugh unit and of weight loss during storage at 25°C, and it was more effective than SAEW or UV alone in maintaining egg internal quality (P < 0.05). The results highlight the promising use of a SAEW and UV combination treatment to improve egg internal quality during storage.

[Transthoracic ultrasonographic features of typical high-resolution computed tomography signs of interstitial lung diseases].

Objective: To investigate the accuracy of bedside transthoracic lung ultrasonography (TLU) in different typical high resolution computed tomography (HRCT) signs of interstitial lung diseases (ILDs). Methods: Fifty patients first diagnosed with ILDs were enrolled from January 2016 to December 2018. There were 21 males and 29 females. The mean age was (56±14) years(rang 42-73 years). TLU was performed in inspiration for the characters of A-lines and B-lines as well as pleural at anterior, lateral and dorsal chest walls, respectively. HRCT was selected at three levels according to the upper, middle, and lower lung fields. The range of each level needing to be evaluated corresponded to the TLU scanning field one by one, and recording the signs of HRCT. Early change of ILDs was definite as the HRCT score was no more than 1 and no honeycomb was present. The correlation between A-lines, B-lines, pleural abnormal and HRCT signs was evaluted. Spearman's correlation coefficient was used to evaluate the relationship between B-lines and HRCT score. Results: The sensitivity and specificity of A-lines for HRCT normality were 83.9% and 84.9%, respectively. Coincidence rate was 84.6%. The sensitivity and specificity of B-lines for HRCT abnormality were 84.9% and 83.9%, respectively. Coincidence rate was 84.6%. Interlobular septal thickening shadow had fewer B-lines and narrower interval than other HRCT signs, while the other HRCT signs had no differences in B-lines. And the sensitivity and specificity of B-lines for detection the early change of HRCT in ILDs were 89.5% and 89.2%, respectively. Coincidence rate was 89.3%. A positive correlation was found between the number of B-lines and HRCT scores (R=0.827, P<0.001), and the width of B-lines and HRCT score (R=0.951, P<0.001). Meanwhile, a negative correlation was found between the interval of B-lines and HRCT score (R=-0.831, P<0.001). The sensitivity and specificity of TLU for HRCT pleural abnormality were 100.0% and 90.0%, respectively. Coincidence rate was 93.6%. Conclusions: TLU showed high sensitivity and specificity in finding interstitial changes of the lung. It gives a new view on the diagnostic possibilities of ILDs and may be used to evaluate the severity and the therapeutic effect of treatment. However, TLU could not differentiate HRCT signs of ILDs.

LncRNA DANCR aggravates the progression of ovarian cancer by downregulating UPF1.

OBJECTIVE: To uncover the role of long non-coding RNA (lncRNA) DANCR in aggravating the progression of ovarian cancer (OC) by downregulating UPF1 level. PATIENTS AND METHODS: DANCR level in OC tissues and matched adjacent normal ones was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Its expression level in OC patients with different tumor node metastasis (TNM) staging and either with metastasis, or not, was examined as well. Receiver operating characteristic (ROC) curves were introduced for assessing the prognostic value of DANCR in OC. Subsequently, regulatory effects of DANCR on proliferative and migratory abilities of HO8910 and HEY cells were evaluated. Subcellular distribution of DANCR in OC cells was analyzed. Furthermore, the interaction between DANCR and UPF1 was explored by RNA immunoprecipitation (RIP) and Pearson correlation analysis. Finally, rescue experiments were conducted to clarify the role of DANCR/UPF1 axis in the progression of OC. RESULTS: DANCR was upregulated in OC tissues and cell lines. Its level was higher in OC patients with worse tumor stage and accompanied by metastatic loci. DANCR exerted the potential to serve as a prognostic marker for OC. Overexpression of DANCR accelerated HO8910 and HEY cells to proliferate and migrate. UPF1 was found to be downregulated in OC tissues and negatively correlated to DANCR. DANCR was mainly distributed in the cytoplasm and interacted with UPF1. Overexpression of UPF1 in OC cells partially reversed the promotive effect of DANCR on proliferative and migratory rates. CONCLUSIONS: LncRNA DANCR accelerates the proliferative and migratory abilities of OC cells through negatively regulating UPF1 level, thus aggravating the progression of OC.

P2X3 receptor-mediated visceral hyperalgesia and neuronal sensitization following exposure to PTSD-like stress in the dorsal root ganglia of rats.

BACKGROUND: Patients with posttraumatic stress disorder (PTSD) often share co-morbidity with chronic pain conditions. Recent studies suggest a role of P2X3 receptors and ATP signaling in pain conditions. However, the underlying mechanisms of visceral hyperalgesia following exposure to PTSD-like stress conditions remain unclarified. METHODS: The behavior and hormones relevant for PTSD were studied. Visceromotor responses (VMR) and the abdominal withdrawal reflexes (AWR) to colorectal distention (CRD) were recorded to determine P2X3-receptor-mediated alteration of hyperalgesia following single-prolonged stress (SPS) exposure. Immunofluorescence, Western blotting, and patch-clamp were used. KEY RESULTS: The escape latency, adrenocorticotropic hormone and cortisol were increased on days 7-14. Visceromotor responses and AWR was reduced at day 1 in SPS rats but increased to higher levels than in controls after exposure to day 7. Intrathecal administration of the P2X3-receptor antagonist TNP-ATP abolished the CRD response. Based on immunofluorescence and Western blotting analysis, SPS-treated rats exhibited reduced P2X3 expression in dorsal root ganglia (DRG) after day 1 compared with controls. P2X3 expression in DRG was enhanced on day 7 after SPS and the increase of the P2X3 expression was maintained on day 14 and 21 compared with controls. The P2X3-receptor agonist α,ß-me ATP (10 µM) induced a fast desensitizing inward current in DRG neurons of both control and SPS-treated rats. The average peak current densities in SPS-treated group were increased 3.6-fold. TNP-ATP (100 nM) markedly blocked all fast α,ß-me ATP-induced inward currents in the DRG neurons both in control and SPS-treated rats. CONCLUSIONS & INFERENCES: The data indicate an important role of P2X3 signaling in visceral hyperalgesia following PTSD-like stress.

[Inactivation of PMS2 gene by promoter methylation in nasopharyngeal carcinoma].

Objective: To investigate the inactivation of PMS2 gene mediated by promoter methylation and its regulatory mechanism in nasopharyngeal carcinoma (NPC). Methods: Fifty-four NPC tissues, 16 normal nasopharyngeal epithelia (NNE), 5 NPC cell lines (CNE1, CNE2, TWO3, HNE1 and HONE1) and 1 normal nasopharyngeal epithelial cell line (NP69) were collected.Methylation-specific PCR (MSP) was used to detect the PMS2 promoter methylation, semi-quantitative reverse transcription PCR (qRT-PCR) was applied to determine its mRNA expression, and immunohistochemistry (IHC) was used to detect the protein expression of PMS2. The expressions of PMS2 mRNA in CNE1 and CNE2 cells before and after treated with methyltransferase inhibitor 5-aza-2-deoxycytidine were analyzed by qRT-PCR. The impact of methylation and demethylation on the mRNA expression of PMS2, and the association of mRNA and protein expression of PMS2 with clinicopathological features of nasopharyngeal cancer were analyzed. Results: Methylation of PMS2 gene was detected in all of the five NPC cell lines, but not in normal nasopharyngeal epithelial NP69 cells. The methylation rate of PMS2 gene in NPC tissues was 63% (34/54), significantly higher than that of the normal nasopharyngeal epithelia (0/16, P<0.001). The expression levels of PMS2 mRNA and protein were significantly down-regulated in the 54 NPC tissues when compared with those in the 16 NNE tissues (P<0.001), and were also significantly lower in the 34 methylated NPC tissues than those in the 20 unmethylated NPC tissues (P<0.001). After treatment with 5-aza-2-deoxycytidine, the expression of PMS2 mRNA was restored in the CNE1 and CNE2 cells.However, the expressions of PMS2 mRNA and protein were not significantly correlated with patients' age, gender, TNM stage, histopathologic type or lymph node metastasis (P>0.05 for all). Conclusions: Promoter methylation-mediated inactivation of PMS2 gene participates in carcinogenesis and development of NPC. PMS2 may be a candidate tumor suppressor in the treatment for patients with inactivation of PMS2 promoter methylation.

[Role of magnetic resonance imaging in the diagnosis of juvenile dermato-myositis and polymyositis in Chinese children].

Objective: To evaluate the utility of magnetic resonance imaging (MRI) in diagnosis of juvenile dermatomyositis and polymyositis (JDM-PM) in children. Method: Fifty-four patients with JDM-PM in the active stage were enrolled in the study group. Twelve patients with benign acute childhood myositis and forty patients with juvenile idiopathic arthritis (JIA) complicated with myositis were enrolled as controls. MRI imaging of thighs was performed in all patients, fast spin echo T1WI, T2WI, and STIR were obtained in all patients.Muscle biopsy was performed in 41/54 patients with JDM-PM. We compared the value of MRI in diagnosis of JDM-PM with muscle biopsy, electromyography and serum aspartate transaminase (AST), alanine transaminase (ALT), creatine kinase (CK), isoenzyme of creatine kinase (CKMB), lactate dehydrogenase (LDH), hydroxybutyrate dehydrogenase (HBDH) levels. Continuous normally distributed variables were reported as means and continuous non-normally distributed variables as median. Chi-square test and Fisher exact test were used to test differences between MRI and other categorical variables. Result: A total of 54 patients were included. Twenty-seven patients were male and the others were female. Average age of the patients was (7.1±3.5) years (2-13 years); 45(83%) paitests were JDM cases and 9(17%) patients had JPM. All patients had MRI examination. Of the 54 patients, 53 had multiple myositis; 10 out of 50 (19%) patients received second MRI after treatment, 6 out of 10 patients had normal findings, 4 patients showed obviously improved images; 41 out of 54 patients underwent muscle biopsy; 22 out of 41 patients had inflammatory cells infiltration and muscle fiber degeneration. The results of the muscle enzyme tests are as follows: 27 (50%) patients had elevated AST, 24 (44%) patients had elevated ALT, 22 (41%) patients had elevated CK, 18(33%) patients had elevated CKMB, and LDH rose in 30 (56%) patients, HBDH rose in 28(52%) patients. These results suggested that muscle MRI was more sensitive than muscle biopsy and muscle enzyme tests in diagnosis of JDM-PM. Conclusion: Patients with JDM-PM showed diffuse patchy hyperintense signals on T2WI of their thighs. MRI may be a sensitive, reliable, and noninvasive tool for clinical diagnosis and theraputic evaluation of JDM-PM.

[Oxidative damage related to PM2.5 exposure in human embryonic stem cell-derived fibroblasts].

OBJECTIVE: To study the oxidative damage of PM2.5 in human embryonic stem cells (EBf-H9 cells), and to provide a theoretical basis for revealing the adverse health effects of PM2.5 and the potential mechanisms, and also to provide a new alternative cell model for PM2.5 risk assessment. METHODS: EBf-H9 cells were cultured with 0.00 (the constrast group) 3.91, 7.81, 15.63, 31.25, 62.50, 125.00 µg/cm(2) of PM2.5. CCK-8 kit was used to determine the survival rate of cells exposed to PM2.5 for 6 h. DCFH-DA probe was used to detect the total ROS content of cells exposed to PM2.5 for 1 h. The activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px), and the content of lipid peroxides such as malondialdehyde (MDA) in cells exposed to PM2.5 for 6 h were detected by using the commercial kits. ANOVA model analyzed the statistical significance from the different concentration group. RESULTS: The cytotoxicity results showed that the cell survival rate was decreased gradually with the increase of the concentrations of PM2.5, and the half inhibitory concentration (IC50) was 83.01 µg/cm(2). When the exposure concentration was 3.91, 7.81, 15.63, 31.25, 62.5 µg/cm(2), after exposure of PM2.5 1 h, the ROS florescence was 27.12±0.21, 54.03±0.50, 60.93±0.08, 61.36±1.00, 68.21±0.93, 78.27±1.26 (compared to control group 27.12±0.21, all P level<0.01). After exposure of PM2.5 6 h, the activities of T-SOD was (9.78±0.28), (8.59±0.22), (8.90±0.33), (7.46±0.71), (4.21±0.17) U/mg protein (F=98.881, compared to control group (11.77±0.63) U/mg protein, all P level<0.01). The activities of GSH-Px was (181.59±3.65), (153.33±1.69), (168.74±2.22), (81.56±0.56), (48.62±2.13) U/mg protein (compared to control group (273.90±6.50) mU/mg protein, all P level<0.01). And the content of MDA was (0.38±0.03), (0.43±0.09), (0.47±0.09), (0.65±0.10), (0.70±0.12) nmol/mg protein (compared to control group (0.27±0.02) nmol/mg protein, all P level<0.05). CONCLUSION: PM2.5 exposure can decrease EBf-H9 cells viability, and improve the levels of lipid peroxidation. It may be due to induce EBf-H9 cells to increase the production of ROS and to make the cells appear oxidative stress, which lead to oxidative damage to cells. The present study reveals the mode of action of PM2.5 in terms of oxidative damage to EBf-H9 cells. It is also indicated that the cells may be a new alternative cell model for PM2.5 risk assessment.

[PM2.5 induces oxidative damage and affects nuclear factor-erythroid 2 related factor 2 pathway in human umbilical vein endothelial cells].

OBJECTIVE: To assess the oxidative damage after exposure to fine particulate matter (PM2.5) in human umbilical vein endothelial cells (HUVECs) and to explore the influence of the Nrf2 pathway. METHODS: HUVECS were exposed to different concentrations of PM2.5 as follows: 0.000 (control), 0.004, 0.039, 0.391, 1.950, 3.910, 7.810, 15.600, 31.250, 62.500, 125.000 and 250.000 µg/cm(2). After 24 h, cell viability was measured by the CCK-8 method. In a separate experiment, HUVECs were exposed to 0 (control), 1.95, 3.91, 7.81, 15.63 or 31.25 µg/cm(2) of PM2.5. The level of cellular reactive oxygen species (ROS) was detected with an H2-DCFDA fluorescence probe after 1h and 3 h exposure. After 24 h exposure, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx), and malondialdehyde (MDA) content were detected by colorimetry. Western blot was used to estimate the expression levels of Nrf2 and NQO1 in total protein. RESULTS: HUVEC viability was reduced in a concentration-dependent manner by PM2.5. Compared with controls (100% viability), the viability of the 250 µg/cm(2) group was (38.18±6.68)% (P<0.05). Substantial accumulation of ROS occurred in HUVEC after 1 h and 3 h exposure to PM2.5. After 24 h exposure to 0, 1.95, 3.91, 7.81, 15.63 and 31.25 µg/cm(2) of PM2.5, SOD activity decreased concentration-dependently to (26.25±1.76), (24.99±1.81), (24.25±0.49), (22.07±1.13), (21.03±0.43) and (19.37±0.84) U/mg protein, respectively, (F=13.95, P<0.001). GPx activity decreased in a concentration-dependent manner to (25.63±1.33), (24.40±2.20), (22.85±2.46), (20.98±1.95), (20.17±1.86) and (18.69±3.11) mU/mg protein, respectively (F=4.26, P=0.019), whereas MDA increased concentration-dependently to (1.11±0.07), (1.12±0.07), (1.17±0.05), (1.49±0.01), (1.95±0.08) and (2.37±0.08) nmol/mg protein, respectively, (F=186.37, P<0.001). Compared with the control Nrf2, NQO1 protein levels (1.00±0.27, 1.00±0.33), 15.63 µg/cm(2) group (2.38±0.44, 1.78±0.20) were enhanced (P<0.05). CONCLUSION: These results demonstrate that PM2.5 can lead to oxidative damage to HUVEC in a concentration-dependent manner. Protein levels of Nrf2 and NQO1 were enhanced at high concentrations of PM2.5, and this mechanism may be related to increases in cellular ROS induced by PM2.5.

[TAK1 promotes epithelial-mesenchymal transition of lens epithelial cells].

OBJECTIVE: Transforming growth factor-ß-activated kinase-1 (TAK1) is thought to play a key role in the initiation of Smad-independent TGF-ß signaling. This study investigated the role of TAK1 in the epithelial-mesenchymal transition (EMT) lens epithelial cells. METHODS: TAK1 was overexpressed in the HLE B-3 cell line by transfecting TAK1-pcDNA3 and TAK1-binding protein 1 (TAB1)-pcDNA3 plasmids. The expression levels of TAK1, phospho-TAK1, E-cadherin, and fibronectin were detected by Western blot analysis and immunocytofluorescence to analyze the effects of overexpression. The levels of α-SMA and type I collagen were analyzed by real-time PCR. Quantitative data were analyzed by Student's t test or one-way analysis of variance (ANOVA) (multiple comparisons using LSD test). RESULTS: Western blot analysis showed in the TAK1-pcDNA3 plasmids group, expression of TAK1 proteins (1.00±0.03) with a maximum upregulation of approximately 80% at 24 h than it was in the control group (0.19±0.09)(t=8.02, P< 0.01); Western blot analysis showed in the TAB1-pcDNA3 plasmids group, expression of TAB1 proteins (1.00±0.02) with a maximum upregulation of approximately 78% at 24 h than it was in the control group (0.22±0.08)(t=7.63, P<0.01). The levels of E-cadherin/Beta-actin had significant differences among control, overexpression of TAK1 together with TAB1, overexpression of TAK1, and overexpression of TAB1 (1.00±0.02, 0.12±0.03, 0.98±0.09, 0.92±0.08;F=31.03, P<0.01). The levels of fibronectin/Beta-actin had significant differences among control, overexpression of TAK1 together with TAB1, overexpression of TAK1, and overexpression of TAB1 (0.11±0.03, 1.00±0.05, 0.16±0.04, 0.21±0.05;F=35.12, P<0.01). Overexpression of TAK1 with TAB1 resulted in upregulated expression of fibronectin, and downregulated expression of E-cadherin. The expression of E-cadherin was increased and the expression of fibronectin was decreased by TAK1 siRNA and TAK1 chemical inhibitors in the presence of TGF-ß2. CONCLUSION: These data reveal that TAK1 can induce the EMT of HLE cells, and the inhibition of TAK1 phosphorylation may be a potential novel therapeutic target for the prevention and treatment of posterior capsule opacification. (Chin J Ophthalmol, 2016, 52: 278-284).