A combined diagnostic model based on circulating tumor cell in patients with solitary pulmonary nodules.

BACKGROUND: Although many prediction models in diagnosis of solitary pulmonary nodules (SPNs) have been developed, few are widely used in clinical practice. It is therefore imperative to identify novel biomarkers and prediction models supporting early diagnosis of SPNs. This study combined folate receptor-positive circulating tumor cells (FR+ CTC) with serum tumor biomarkers, patient demographics and clinical characteristics to develop a prediction model. METHODS: A total of 898 patients with a solitary pulmonary nodule who received FR+ CTC detection were randomly assigned to a training set and a validation set in a 2:1 ratio. Multivariate logistic regression was used to establish a diagnostic model to differentiate malignant and benign nodules. The receiver operating curve (ROC) and the area under the curve (AUC) were calculated to assess the diagnostic efficiency of the model. RESULTS: The positive rate of FR+ CTC between patients with non-small cell lung cancer (NSCLC) and benign lung disease was significantly different in both the training and the validation dataset (p < 0.001). The FR+ CTC level was significantly higher in the NSCLC group compared with that of the benign group (p < 0.001). FR+ CTC (odds ratio, OR, 95% confidence interval, CI: 1.13, 1.07-1.19, p < 0.0001), age (OR, 95% CI: 1.06, 1.01-1.12, p = 0.03) and sex (OR, 95% CI: 1.07, 1.01-1.13, p = 0.01) were independent risk factors of NSCLC in patients with a solitary pulmonary nodule. The area under the curve (AUC) of FR+ CTC in diagnosing NSCLC was 0.650 (95% CI, 0.587-0.713) in the training set and 0.700 (95% CI, 0.603-0.796) in the validation set, respectively. The AUC of the combined model was 0.725 (95% CI, 0.659-0.791) in the training set and 0.828 (95% CI, 0.754-0.902) in the validation set, respectively. CONCLUSIONS: We confirmed the value of FR+ CTC in diagnosing SPNs and developed a prediction model based on FR+ CTC, demographic characteristics, and serum biomarkers for differential diagnosis of solitary pulmonary nodules.

Comparison of metabolome profiles in zebrafish (Danio rerio) intestine induced by polystyrene microplastics with different sizes.

Microplastics (MPs) are widespread in aquatic environments. They could induce intestinal toxicity in the fish. However, research on the metabolic toxicity of polystyrene microplastics (PS-MPs) with different particle sizes to the zebrafish intestine is still limited. Here, metabolomics using ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was applied to characterize the metabolic disorders in zebrafish intestine after exposure to 500 µg/L PS-MPs with different sizes (100 nm, 5 µm, and 200 µm) for 21 days. Results showed that the 100 nm PS-MPs group increased glutathione content. A total of 35, 165, and 87 metabolites were significantly altered in zebrafish intestines of 100 nm, 5 µm, and 200 µm groups under positive ion mode, respectively. In comparison, 31, 115, and 45 metabolites were changed in the 100 nm, 5 µm, and 200 µm groups under negative ion mode, respectively. Metabolic pathway analysis indicated that carbohydrate metabolism, amino acid metabolism, and nucleotide metabolism were changed in all three groups. The greatest changes were found in the 5 µm group. Moreover, treatment with micro-sized PS-MP groups specifically changed lipid metabolism, which might be related to pathogenic bacteria (Streptococcus and Moraxella). In the 100 nm PS-MP group, S-adenosyl-L-methionine (SAM) was found to be markedly related to the intestinal microbiota. SAM level was significantly increased, which might account for the elevated glutathione content. To sum up, the mechanisms of nano-sized MPs (oxidative stress) and micro-sized MPs (lipid metabolism disorder) were distinct. This study provides novel insight into the toxicity mechanism of MPs in the zebrafish intestine.

Exploring the mechanism of miR320a in regulating PDL1 upon lung cancer pathogenesis.

The tumour suppressive role of miRNA320a is observed in many cancer types like in colon, lung, breast, and osteosarcoma but it is inversely reported in prostate cancer and in MPM cell lines. miRNA320a targets programmed death ligand 1 (PDL1) negatively in many cancer types and recently in Malignant pleural mesothelioma. In this background it is important to understand the regulatory mechanism of miRNA320a in determining PDL1 expression in different pathological stages of lung cancer. Histology was used to grade the initial and advanced stage of lung cancer following carcinogenic injection. Immunohistochemistry and Western blotting technique were used to analyse PDL1 protein expression. In-situ hybridization was used to determine miRNA320a signals. Initially, using the chemical carcinogen Diethylnitrosamine (25 µg/g), we successfully initiate initial and advanced stage of lung cancer following 6 months and 9 months of carcinogenic injection. The formation of initial and advanced stage of lung cancer is confirmed through histopathological changes which show neoplastic appearance in initial lung cancer and appearance of more mitotic cells along with tissue hardness in the advanced lung cancer stages. In miRNA320a blocked tissue the cancer condition becomes worse with decreased tissue elasticity along with more proliferative cells. Immunohistochemistry and Western blotting studies show that PDL1 is overexpressed in the advanced stages rather than in initial lung cancer because the expression of miRNA320a is overexpressed in initial stages but restricted in advanced stages of lung cancer. miRNA32a blocking studies confirm that miRNA320a expression act as a tumour suppressor that directly controls PDL1 expression that lack of miRNA320a enhances PDL1 expression as well as it triggers lung cancer advances. In summary, miRNA320a possess tumour suppressor function that limits PDL1 expression in initial lung cancer but its control over PDL1 suppression is lost once miRNA320a is downregulated in advanced stage of lung cancer.

Clinical use of 18F-FDG PET/CT in the differential diagnosis of patients with primary and secondary adenoid cystic carcinoma of the lung: a retrospective cohort study.

Background: Adenoid cystic carcinoma (ACC) of the lung (ACCL) is a rare malignancy and includes primary ACCL (PACCL) and secondary ACCL (SACCL) metastasized from the ACC of the head and neck. Fluorine-18-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) has been shown to be useful in the differential diagnosis between primary and metastatic lung lesions. This study retrospectively investigated the role of 18F-FDG PET/CT in combination with clinicopathological findings in the management of patients with primary or secondary ACCL. Methods: Clinicopathological characteristics and 18F-FDG PET/CT metabolic parameters of 29 patients with PACCL and 11 patients with SACCL with pathological confirmation as gold standard were retrospectively collected. The association between PET/CT metabolic parameters and clinicopathological features was explored. Receiver operating characteristic (ROC) curve analysis was used to determine the optimal maximum standard uptake value (SUVmax) cut-off value to distinguish PACCL from SACCL. The prognostic value of PET/CT metabolic parameters and clinicopathological features was evaluated by Cox regression analysis. Results: SACCL patients more often presented with multiple ACC lesions in the peripheral lung (81.8% vs. 17.2%, P<0.05) and tended to be asymptomatic compared to patients with PACCL (72.7% versus 27.6%, P<0.05). The SUVmax was significantly higher in PACCL patients compared to SACCL patients (median 4.4 vs. 2.8, P<0.05). Furthermore, at a cut-off value of 3.2 for SUVmax, the sensitivity and specificity of 18F-FDG PET/CT in distinguishing PACCL from SACCL were 82.8% and 72.7%, respectively. Higher SUVmax of ACCL was observed in patients with clinical symptoms, fewer ACC lesions, or larger tumor size (P<0.05). The median PFS of ACCL patients was 71.1 months, and the 12- and 24-month PFS rates were 96.1% and 91.6%, respectively. Univariate Cox regression analysis showed that SACCL from metastasis, lesions located at the peripheral lung or multiple ACC lesions present in the lung, were associated with poorer PFS (P<0.05). However, multivariate Cox regression analysis showed that none of the variables examined were independent predictors of prognosis. Conclusions: The use of 18F-FDG PET/CT in combination with assessment of clinicopathological features was helpful in distinguishing PACCL from SACCL, which could provide guidance for clinical decision making.

Exosomes derived from M2 type tumor-associated macrophages promote osimertinib resistance in non-small cell lung cancer through MSTRG.292666.16-miR-6836-5p-MAPK8IP3 axis.

BACKGROUND: Osimertinib resistance limits the treatment of epidermal growth factor receptor-(EGFR)-mutated non-small-cell lung carcinoma (NSCLC). The mechanisms of osimertinib resistance need to be elucidated to determine alternative treatment strategies. This study explores the role of M2 type tumor-associated macrophage (TAM)-derived exosomal MSTRG.292666.16 in osimertinib resistance, and its related competing endogenous RNA (ceRNA) mechanism. METHODS: M2 type TAMs were induced with 200 ng/mL phorbol 12-myristate 13-acetate, 20 ng/mL IL-4 and IL-13, and M2 type macrophage markers were measured by RT-qPCR. Next, the exosomes were isolated and characterized. Tumor formation in nude mice was conducted using H1975 cells under different treatment conditions. Small RNA sequencing was performed on exosomes derived from sensitive and resistant plasma, and ceRNA networks were constructed. Fluorescence in situ hybridization was used to observe the localization of MSTRG.292666.16, and a ceRNA network (MSTRG.292666.16-miR-6836-5p-MAPK8IP3) was selected for further validation. RESULTS: M2 type TAMs, and M2 type TAM-derived exosomes were successfully induced and isolated. Nude mice results showed that M2 type TAM-derived exosomes and MSTRG.292666.16 overexpression significantly increased tumor volume after administration of osimertinib for 4 weeks. M2 type TAMs were found in the resistant plasma, and MSTRG.292666.16 localized in the cytoplasm of H1975 cells. In addition, the genes in the ceRNA networks were significantly enriched in eight GO terms and seven KEGG pathways, including the MAPK signaling pathway. Subsequently, the levels of MSTRG.292666.16 and MAPK8IP3 significantly increased in both resistant plasma-derived exosomes and M2 type TAM-derived exosomes, while miR-6836-5p levels were significantly reduced. Finally, MSTRG.292666.16, miR-6836-5p, and MAPK8IP3 were part of the same network. CONCLUSIONS: M2 type TAM-derived exosomes promoted osimertinib resistance in NSCLC by regulating the MSTRG.292666.16/miR-6386-5p/MAPK8IP3 axis.

Heterogeneity effects of nanoplastics and lead on zebrafish intestinal cells identified by single-cell sequencing.

Plastic particles in water environment can adsorb heavy metals, leading to combined toxicity on aquatic organisms. However, current conclusions are mostly obtained based on cell population-average responses. Heterogeneity effects among cell populations in aquatic organisms remain unclear. This study firstly analyzed the basic toxic effects of 20 µg L-1, 200 µg L-1 100 nm polystyrene nanoplastics (PS-NPs), 50 µg L-1 lead (Pb), and their combined exposures on zebrafish intestine. Results found that combined exposure of 200 µg L-1 PS-NPs and 50 µg L-1 Pb induced highest MDA, 8-OHdG, and TNF-α levels. Thus 200 µg L-1 PS-NPs, 50 µg L-1 Pb and their combined exposures were chosen to analyze the heterogeneity effects on zebrafish intestine cells by single-cell RNA sequencing. A total of 38,640 zebrafish intestinal cells were obtained and identified as seven cell populations, including enterocytes, macrophages, neutrophils, B cells, T cells, enteroendocrine cells, and goblet cells. 200 µg L-1 PS-NPs exposure had the greatest influence on macrophages, while Pb exposure mostly influenced enterocytes. Results of MDA, 8-OHdG, and TNF-α analyses indicated that 20 µg L-1 and 200 µg L-1 PS-NPs increased the Pb toxicity. However, the scRNA-seq showed that the synergistic effects did not exist in most cell populations, except for goblet cells. Co-exposure of 200 µg L-1 PS-NPs and Pb caused similar transcriptome profiles with 200 µg L-1 PS-NPs exposure in macrophages, which changed immunological recognition and apoptosis processes. The Pb exposure influenced the macrophages by direct cytotoxicity. However, the Pb alone and combined exposures induced similar toxicities in the enterocytes, including the generation of oxidative stress and abnormality of lipid metabolism. This study shows the scRNA-seq is a powerful method to identify the target cell populations and corresponding toxic effects during combined exposure of pollutants.

Application of neck ultrasound in the diagnosis of sarcoidosis.

OBJECTIVE: To explore the significance of neck ultrasound (NUS) combined with contrast-enhanced ultrasound (CEUS) in the diagnosis of sarcoidosis. METHODS: 88 patients with evidence of intrathoracic lymphadenopathy and suspected sarcoidosis with enlarged cervical lymph nodes underwent NUS, CEUS, fine-needle aspiration and core needle biopsy when technically feasible were retrospectively analyzed in this study. Seven characteristics such as enhanced mode (EM), resolution time, color Doppler flow imaging (CDFI), fading time, peaking state-uniformity, strengthen the area and symmetry were considered to perform the logistic regression model. RESULTS: Of 88 patients included in this study, sarcoidosis was accounted in 20 cases, tuberculosis in 23 cases, malignancy in 22 cases and inflammatory lymph node in 23 cases. There were statistically significant differences in symmetry, lymphatic hilum, homogeneity, CDFI pattern and elasticity score between the sarcoidosis and non-sarcoidosis groups via NUS. Similarly, we also acknowledged a statistically significant differences in EM, homogeneity, presence or absence of necrosis between the sarcoidosis and non-sarcoidosis groups via CEUS to further group the non-sarcoidosis into tuberculosis, malignancy or inflammatory disorder. The percentage correction of prediction was 90% (18/20). CONCLUSION: NUS combined with CEUS has characteristic features in sarcoidosis with cervical lymph node involvement, which is helpful for its diagnosis and differential diagnosis. The binary classification model of NUS combined with CEUS features can help differentiate sarcoidosis from non-sarcoidosis groups.

The impact of gender and working hours on pulmonary physician burnout.

BACKGROUND: Physician burnout has attracted an increasing amount of attention as a global public health problem. It is associated with negative clinical and organizational outcomes, including lower patient satisfaction, patient safety, and quality of care. The work of pulmonary physicians involves high levels of stress, putting them at higher risk of burnout. This study aimed to evaluate the personal and professional characteristics associated with burnout among pulmonary physicians. METHODS: Between March 2020 and May 2020, a survey to evaluate burnout and career satisfaction among pulmonary physicians was conducted in Shanghai Pulmonary Hospital. The survey collected information on physician characteristics, career satisfaction, and burnout. The Maslach Burnout Inventory (MBI), the gold standard tool for measuring burnout, was used to measure burnout symptoms. Self-reported professional satisfaction in relation to career, specialty, and work-life balance was recorded. RESULTS: Of 130 pulmonary physicians contacted, 125 (96.1%) completed full-length surveys. Ninety-one (72.8%) of these 125 physicians were female. The survey participants had a median age of 40 years. Of the respondents, 48.8% had at least one symptom of burnout according to high emotional exhaustion (EE) score and/or high depersonalization (DP) score. Univariable analysis showed female gender, the number hours worked per week, the number of nights on call per month, and the number of calls per night shift to be factors associated with burnout. Gender and hours worked per week were independent factors associated with burnout in multivariate analysis. CONCLUSIONS: The MBI was able to characterize burnout experienced by pulmonary physicians in our hospital. Gender and the number of hours worked per week were the significant factors independently associated with burnout among the physicians in our study.

Single-Cell Sequencing Reveals Heterogeneity Effects of Bisphenol A on Zebrafish Embryonic Development.

The embryonic period is a sensitive window for bisphenol A (BPA) exposure. However, embryonic development is a highly dynamic process with changing cell populations. The heterogeneity effects of BPA on fish embryo cells during development remain unclear. We applied single-cell RNA sequencing to analyze the impact of BPA exposure on transcriptome heterogeneity of 64 683 cells from zebrafish embryos at 8, 12, and 30 h postfertilization (hpf). Thirty-eight cell populations were identified and gene expression profiles of 16 cell populations were significantly altered by BPA. At 8 hpf, BPA mainly influenced the outer layer cell populations of embryos, such as neural plate border and enveloping layer cells. At 12 and 30 hpf, nervous system formation and heart morphogenesis were disturbed. The altered differential processes of the neural plate border, neural crest, and neuronal cells were found to lead to increased neurogenesis in zebrafish larvae. In the forebrain, midbrain, neurons, and optic cells, pathways related to cell division and DNA replication and repair were altered. Moreover, BPA also changed transforming growth factor (TGF) ß signaling and heart tube morphogenesis in heart cells, leading to a decreased heartbeat in zebrafish larvae. Our study provides a comprehensive understanding of BPA toxicity on fish embryonic development at a single-cell level.

Influence of the digestive process on intestinal toxicity of polystyrene microplastics as determined by in vitro Caco-2 models.

The digestive tract is an important target organ for microplastics (MPs). However, little is known about the effects of digestive treatment on the intestinal toxicity of MPs. In this study, an in vitro digestive process was applied to transform 100 nm and 5 µm polystyrene microplastics (PS-MPs). Intestinal toxicities of original PS-MPs and transformed PS-MPs (t-PS-MPs) were determined using an in vitro Caco-2 monolayer model. Results showed that the digestive process did not alter the chemical constitution of PS-MPs, but formed a corona on the surface of PS-MPs. The 100 nm PS-MPs showed higher intestinal toxicity than 5 µm PS-MPs. Digestive treatment relieved cytotoxicity and transport function disorder of the Caco-2 monolayer induced by the original PS-MPs. Moreover, the combined toxicities of PS-MPs and arsenic were also decreased by digestive treatment. However, the in vitro digestive process increased the proinflammatory effects of PS-MPs. The formation of a corona on the PS-MP surface, which lead to a change in size, Zeta potential, and adsorbed compounds, might induce the above influence of digestive treatment. Our study suggests that direct cytotoxicity assays of PS-MPs might misestimate their intestinal effects, which provide new lights to the toxicity evaluation of PS-MPs by oral exposure.

Clinicopathological and Prognostic Significance of EML4-ALK Rearrangement in Patients with Surgically Resected Lung Adenocarcinoma: A Propensity Score Matching Study.

OBJECTIVE: The echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK) fusion gene is a key oncogenic driver in non-small cell lung cancer (NSCLC). This study analyzed the clinicopathological characteristics and prognostic significance of EML4-ALK fusion gene in patients with surgically resected adenocarcinoma. METHODS: The clinicopathological characteristics of 1056 consecutive patients with surgically resected stage I-IIIA adenocarcinoma were collected from February 2014 to October 2014, and EML4-ALK rearrangement was detected using real-time polymerase chain reaction (RT-PCR) technology. To compare the imaging and pathological features, a propensity score matching (PSM) method was performed. The follow-up information was collected to evaluate the long-term outcomes of patients with EML4-ALK rearrangement. RESULTS: The prevalence of EML4-ALK rearrangement was 6.6% in 1056 consecutive patients. A total of 70 EML4-ALK-positive and 210 EML4-ALK-negative patients were identified after PSM. Imaging and pathological analyses showed that EML4-ALK rearrangement was significantly associated with less ground-glass opacity (GGO) (adjusted OR=1.38, 95% CI=1.03-1.85, Ptrend =0.029) and higher prevalence of non-invasive mucinous adenocarcinoma mucin-laden adenocarcinomas (non-IMA MLA, adjusted OR=6.79, 95% CI=2.69-17.17, P<0.001). EML4-ALK rearrangement was found to be an unfavorable prognostic factor for disease-free survival (DFS) in female patients (HR=2.26, 95% CI=1.13-4.53, P=0.021). CONCLUSION: Our results suggest that adenocarcinomas harboring EML4-ALK fusion gene exhibit specific radiological and pathological characteristics compared with EML4-ALK-negative adenocarcinomas. In female patients, EML4-ALK rearrangement was associated with shorter DFS.

Single-Cell RNA Sequencing Reveals Size-Dependent Effects of Polystyrene Microplastics on Immune and Secretory Cell Populations from Zebrafish Intestines.

Microplastics (MPs) as widespread contamination pose a high risk for aquatic organisms. However, the current understanding of MP toxicity is based on cell population-averaged measurements. Our aim was to gain a comprehensive understanding of the size-dependent effects of polystyrene MPs (PS-MPs) on intestinal cell populations in zebrafish and characterize the interplay of MPs, intestinal cells, and intestinal microbiota. Here, we used single-cell RNA sequencing to determine the transcriptome heterogeneity of 12 000 intestinal cells obtained from zebrafish exposed to 100 nm, 5 µm, and 200 µm PS-MPs for 21 days. Eight intestinal cell populations were identified. Combined with changes in intestinal microbiota, our findings highlight a previously unrecognized end point that all three sizes of PS-MPs induced dysfunction of intestinal immune cells (including effects on phagosomes and the regulation of immune system processes) and increased the abundance of pathogenic bacteria. However, only 100 nm PS-MPs altered the expression of genes related to phagocyte-produced reactive oxygen species (ROS) generation and increased mucus secretion by secretory cells. Microsize PS-MPs specifically changed the lysosome (5 µm) and cell surface receptor signaling (200 µm) processes of the macrophages. Our findings pinpoint to cell-specific and size-dependent responses to PS-MPs in fish intestine, which can provide a reference for future study directions.

Geranylgeranylacetone ameliorates lung ischemia/reperfusion injury by HSP70 and thioredoxin redox system: NF-kB pathway involved.

Geranylgeranylacetone (GGA) has been clinically used as an anti-ulcer drug. In the present study, we explored the protective effects of GGA on lung ischemia/reperfusion injury (IRI) and the underlying mechanism. The results demonstrated that GGA ameliorated the lung biochemical and histological alterations induced by IRI, which was reversed by HSP70 inhibition. To further explore the mechanism of GGA action, we focused on NF-kB and thioredoxin (Trx) redox system. It was shown that GGA induced the HSP70 and Trx-1 expression, NF-kB nuclear translocation and activated thioredoxin reductase (TrxR). The Trx-1 expression and TrxR activity was suppressed by HSP70 and NF-kB inhibition, while the nuclear NF-kB p65 expression was suppressed by HSP70 inhibitor. These results indicated that GGA may protect rat lung against IRI by HSP70 and Trx redox system, in which NF-kB pathway may be involved.

Shorter EGFR Dinucleotide Repeat Length Predicts Better Response of Patients with Advanced Non-small Cell Lung Cancer to EGFR Tyrosine Kinase Inhibitor.

The purpose of this study is to evaluate the association between intron 1 CA-repeat polymorphisms of the epidermal growth factor receptor gene (EGFR) and the clinical outcome of Chinese patients with advanced non-small cell lung cancer (NSCLC) treated with EGFR tyrosine kinase inhibitors (TKIs). We genotyped the intron 1 CA-repeat genetic polymorphisms of EGFR in 84 Chinese patients with NSCLC. The relationship between the length of the CA repeats and EGFR mutations in exons 18-21 in the 84 patients was elucidated. We then analyzed the association between the length of the CA repeats and the clinical outcome of EGFR-TKI-treated patients with NSCLC. EGFR mutations in exon 19 were significantly associated with shorter CA repeats. Patients with shorter CA repeats had a significantly longer progression-free survival with EGFR-TKI treatment than those with longer CA repeats. Our results suggest that shorter CA repeats in intron 1 of EGFR are associated with EGFR mutations and the clinical outcomes of TKI-treated patients with NSCLC.

[Application of complete mesocolic excision in stage III colon cancer].

UNLABELLED: To evaluate the efficacy of complete mesocolic excision (CME) for stage III colon cancer. METHODS: Clinical data of 100 patients diagnosed as stage III colon cancer in our hospital from July 2011 to July 2013 were analyzed retrospectively. Fifty-four patients in CME group underwent complete mesocolic excision and 46 patients in control group underwent traditional radical surgery. Lymphadenectomy, postoperative specimen quality and short-term clinical efficacy were compared between two groups. RESULTS: The number of gross dissected lymph nodes and positive dissected lymph nodes in CME group were 26.7 ± 2.6 and 4.3 ± 1.4, which were significantly higher than those in control group (22.9 ± 3.7 and 2.8 ± 1.2) (all P<0.01). There was statistical significance in surgical C-class specimens of CME group were found in 45 cases (83.3%), which were significantly higher than those of control group (29 cases, 63.0%) (P<0.05). The postoperative complication rate of two groups was the same without significant difference(both 13.0%, P>0.05). CONCLUSION: CME is safe and effective for stage III colon cancer, which can improve the quality of surgical specimen and increase the number of dissected lymph nodes, but do not elevate the morbidity of postoperative complication.

Mechanisms behind the inhibition of lung adenocarcinoma cell by shikonin.

Shikonin, a natural naphthoquinone isolated from a traditional Chinese medicinal herb, can exert inhibitory effect on tumor cell growth. However, little has been known concerning the effect of shikonin on lung adenocarcinoma cell and underlying mechanisms. In the present study, we investigated the effect of shikonin on the proliferation, cell cycle arrest, and apoptosis in human lung adenocarcinoma cells. We found that shikonin significantly suppressed the proliferation of lung adenocarcinoma cells compared with control in dose- and time-dependent manner (P < 0.05). In the meantime, our results showed that shikonin markedly increased the proportion of A549 cells at stage G1 as well as induced apoptosis in A549 cells. Furthermore, suppressed CCND1 and elevated caspase3 and caspase7 expression levels at mRNA were found in this study, indicating that shikonin may inhibit the growth of lung adenocarcinoma cell by changing cell cycle and promoting cell apoptosis through the regulation of CCND1, caspase3, and caspase7. Although more studies are needed, this study suggests that shikonin has the potential to be used as an anti-cancer agent in the treatment of lung adenocarcinoma.

A mathematical model for treatment-resistant mutations of hiv.

In this paper, we propose and analyze a mathematical model, in the form of a system of ordinary differential equations, governing mutated strains of human immunodeficiency virus (HIV) and their interactions with the immune system and treatments. Our model incorporates two types of resistant mutations: strains that are not responsive to protease inhibitors, and strains that are not responsive to reverse transcriptase inhibitors. It also includes strains that do not have either of these two types of resistance (wild- type virus) and strains that have both types. We perform our analysis by changing the system of ordinary differential equations (ODEs) to a simple single-variable ODE, then identifying equilibria and determining stability. We carry out numerical calculations that illustrate the behavior of the system. We also examine the effects of various treatment regimens on the development of treatment-resistant mutations of HIV in this model.