Objective     To explore the correlation between the quantitative and qualitative features of CT images and the invasiveness of pulmonary ground-glass nodules, providing reference value for preoperative planning of patients with ground-glass nodules. Methods    The patients with ground-glass nodules who underwent surgical treatment and were diagnosed with pulmonary adenocarcinoma from September 2020 to July 2022 at the Third Affiliated Hospital of Kunming Medical University were collected. Based on the pathological diagnosis results, they were divided into two groups: a non-invasive adenocarcinoma group with in situ and minimally invasive adenocarcinoma, and an invasive adenocarcinoma group. Imaging features were collected, and a univariate logistic regression analysis was conducted on the clinical and imaging data of the patients. Variables with statistical difference were selected for multivariate logistic regression analysis to establish a predictive model of invasive adenocarcinoma based on independent risk factors. Finally, the sensitivity and specificity were calculated based on the Youden index. Results     A total of 555 patients were collected. The were 310 patients in the non-invasive adenocarcinoma group, including 235 females and 75 males, with a meadian age of 49 (43, 58) years, and 245 patients in the invasive adenocarcinoma group, including 163 females and 82 males, with a meadian age of 53 (46, 61) years. The binary logistic regression analysis showed that the maximum diameter (OR=4.707, 95%CI 2.060 to 10.758), consolidation/tumor ratio (CTR, OR=1.027, 95%CI 1.011 to 1.043), maximum CT value (OR=1.025, 95%CI 1.004 to 1.047), mean CT value (OR=1.035, 95%CI 1.008 to 1.063), spiculation sign (OR=2.055, 95%CI 1.148 to 3.679), and vascular convergence sign (OR=2.508, 95%CI 1.345 to 4.676) were independent risk factors for the occurrence of invasive adenocarcinoma (P<0.05). Based on the independent predictive factors, a predictive model of invasive adenocarcinoma was constructed. The formula for the model prediction was: Logit(P)=–1.293+1.549×maximum diameter of lesion+0.026×CTR+0.025×maximum CT value+0.034×mean CT value+0.72×spiculation sign+0.919×vascular convergence sign. The area under the receiver operating characteristic curve of the model was 0.910 (95%CI 0.885 to 0.934), indicating that the model had good discrimination ability. The calibration curve showed that the predictive model had good calibration, and the decision analysis curve showed that the model had good clinical utility. Conclusion     The predictive model combining quantitative and qualitative features of CT has a good predictive ability for the invasiveness of ground-glass nodules. Its predictive performance is higher than any single indicator.

This study aims to develop an improved cell screening system for farnesoid X receptor (FXR) agonists based on a dual luciferase reporter gene system. FXR response element (FXRE) fragments from FXR target genes were cloned and inserted into upstream of firefly luciferase (Luc) gene in the plasmid pGL4-luc2P-Hygro. In combination with the internal reference plasmid containing renilla luciferase, a dual luciferase reporter gene system was developed and used for high throughput screening of FXR agonists. After studying the effects of over-expression of RXR, mouse or human FXR, various FXRE fragments, and different ratio of FXR plasmid amount to reporter gene plasmid, induction efficiency of the screening system was optimized by the known FXR agonist GW4064, and Z factor for the system reached 0.83 under optimized conditions. In summary, an improved cell screening system based on double luciferase reporter gene detection system was developed to facilitate the discovery of FXR agonists, where a new enhanced FXRE element was formed by a superposition of multiple FXRE fragments from FXR target genes, instead of a superposition of traditional IR-1 (inverted repeats-1) fragments.
Humans ; Mice ; Animals ; Transcription Factors/genetics* ; DNA-Binding Proteins/genetics* ; Receptors, Cytoplasmic and Nuclear/genetics* ; Genes, Reporter ; Luciferases/genetics*

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1^G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1^G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).
Humans ; NF-kappa B/metabolism* ; Ginsenosides/pharmacology* ; Amyotrophic Lateral Sclerosis/genetics* ; Culture Media, Conditioned/pharmacology* ; Superoxide Dismutase-1 ; Neurodegenerative Diseases ; Neurons/metabolism* ; Apoptosis

Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics* ; Gene Expression Profiling ; Plant Leaves ; Plant Proteins/metabolism* ; Taste ; Tea ; Transcriptome/genetics*

Objective:Lung ultrasound (LUS) has been used in the diagnosis of neonatal respiratory distress syndrome(RDS) successfully, but there have been no multicenter prospective studies to verify its reliability or determine how to grade RDS with LUS findings.This study aimed to discuss the necessity and feasibility of using LUS findings to determine RDS grades through a multicenter prospective study.Methods:Every researcher participated in the National Neonatal Lung Ultrasound Training Course and receiving 3-6 months of lung ultrasound system training at the National Neonatal Lung Ultrasound Training Center.Patients between June 2018 and May 2020 who met the RDS ultrasound diagnostic criteria and had full available clinical data were included in this study.The LUS examination was completed immediately after the patients were admitted to the hospital.Some of them also underwent chest X-ray examination.Arterial blood gas analysis was completed immediately before or after the LUS ultrasound examination.RDS grading was performed according to the LUS findings and whether the patient had serious complications.Results:A total of 275 qualifying cases were included in this study, which included 220 premature infants and 55 full-term infants, and the primary RDS occurred in 117 cases (42.5%), and secondary RDS occurred in 158 cases (57.5%). LUS manifestations of RDS patients can be divided into three categories: (1)A ground-glass opacity sign: which could be found among 50 infants when they were admitted to the hospital (that was, at their first LUS examination). Twenty-eight of these infants were considered to have wet lungs and were not sent for special management on admission, but LUS showed typical snowflake-like lung consolidation within 0.5 to 4 hours.Twenty-two of them were given mechanical ventilation with exogenous pulmonary surfactant; Eighteen cases were controlled within 6-12 hours, but the lung lesions became more severe in the other 4 infants (due to severe intrauterine infection). (2)Snowflake-like lung consolidations: the first LUS on admission showed typical snowflake-like lung consolidation involving areas ranging from 1-2 intercostal spaces to 12 lung divisions in 204 cases.Thirty-eight infants among them the lung consolidation only had involvement of 1-2 intercostal spaces at the time of admission; Fifteen of them received invasive respiratory support and recovered within 4-12 hours.Twelve patients received noninvasive respiratory support; Seven of them recovered, while five cases developed severe lung illness.The remaining 11 patients who were not given any form of ventilator support developed severe conditions within 1-4 hours.Thirty of them showed snowflake signs involving 12 lung regions at admission.The remaining 136 patients had lung consolidation degree between the two degree above condition.(3)Snowflake-like sign with complications: Twenty-one patients had severe complications such as pneumothorax, pulmonary hemorrhage or/and persistent pulmonary hypertension of the newborn or large area atelectasis, etc, although snowflake lung consolidation did not involve all lung regions.Conclusion:(1) LUS is reliable and accurate for diagnosing RDS.RDS has the same characteristics on ultrasound for both preterm and full-term infants, both primary and secondary RDS.(2) To facilitate the management of RDS, it is necessary to classify RDS according to the ultrasound findings and the presence of severe complications.(3) Based on the results of this study, it is recommended that RDS can be divided into mild, moderate and severe degrees.The exact standards for grading are as follows: Mild RDS: the early stage of RDS, in which lung consolidation shows as a ground-glass opacity sign on ultrasound; Moderate RDS: lung consolidation shows a snowflake sign on ultrasound, not all of the lung fields are involved; Severe RDS meets one or more of the following criteria: lung consolidation shows as a snowflake sign on ultrasound and all lung regions are involved, or regardless of its degree and extent, lung consolidation has caused serious complications, such as pulmonary hemorrhage, pneumothorax, persistent pulmonary hypertension of the newborn, or/and a large area of pulmonary atelectasis.

Objective To investigate the expression and significance of microtubule-associated protein 1 light chain 3 (LC3), an autophagy marker protein, in ameloblastomas. Methods Immunohistochemical methods were employed to evaluate the expression of LC3 in104 cases of ameloblastomas and 20 cases of normal oral mucosal tissues. The results were analyzed by a semiquantitative analysis method. Results The reactivity of LC3 in the epithelial cells of ameloblastomas was positive, and the positivity rate was significantly higher than that in normal oral mucosal tissues (P ＜ 0.05). There were no significant differences between the LC3 expression levels with respect to age, gender, or recurrence (P ＜ 0.05). The positivity rate in mandible ameloblastomas was significantly higher than that in maxilla and gingiva ameloblastomas (P ＜ 0.05). The reactivity for LC3 was significantly higher in solid ameloblastomas than that in the other three tissue types (P ＜ 0.05). Conclusion Autophagic activity in ameloblastomas was higher than that in normal oral mucosal tissues. This suggests that autophagy plays an important role during tumorigenesis, and contributes to the local invasion of ameloblastomas.

Objective To evaluate the efficacy of the bacterial lysate Broncho-vaxom on chronic obstructive pulmonary disease(COPD)in elderly patients.Methods A total of 150 patients with stable COPD were randomized into the Broncho-vaxom group(n=78)treated with Broncho-vaxom as add-on to conventional treatment and the control group(n=72)treated with conventional treatment.The number of acute exacerbation of COPD per patient per year,quality of life,lung function,T cell subsets were evaluated for the therapeutic effects.Results After one year of treatment,the number of acute exacerbation per patient per year was lower in Broncho-vaxom group than in control group[(1.44 ± 1.05) times/y vs.(1.82 ± 0.61) times/y,t =2.754,P =0.007].The proportions of acute exacerbationfree patients were higher in Broncho-vaxom group than in control group(20.5％ or 16/78 vs.4.2％ or 3/72,x2 =9.043,P =0.003).There were significant differences in the forced vital capacity(FVC),forced expiratory volume in one second(FEV1),and forced expiratory volume in one second/predicted value ratio(FEV1 ％)between the two groups at 5 different time points(before,at 3,6,9 and 12 months after treatment) (Broncho-vaxom group:F =90.819,50.674 and 51.233,P =0.000;control group:F =84.928,90.654 and 86.117,P =0.000).After treatment,the symptom scores were lower in Broncho-vaxom group than in control group,and the ratios of CD4/CD8 T-cell were increased and level of CD8 T-cell was decreased in Broncho-vaxom group(all P ＜ 0.05).The rate of adverse reactions had no significant difference between Broncho-vaxom group and control group(3/78 or 3.8％ vs.2/ 72 or 2.8％,x2 =0.132,P=0.717).Conclusions The oral administration of Broncho-vaxom for six months can modulate and enhance immune functions,significantly decrease acute exacerbation frequency,improve quality of life and delay the deterioration in lung function in elderly patients with stable COPD.

Objective To evaluate the regulatory role of azithromycin-induced persistent Chlamydia trachomatis (Ct) infection in the apoptosis of Hela229 cells.Methods Hela229 cells were firstly co-cultured with Ct for 22 hours,and then cultured with Dulbecco's modified Eagle's medium (DMEM) containing 0.08 mg/L azithromycin for 26 hours to establish a cell model of persistent Ct infection (persistent infection group).These infected Hela229 cells cultured with azithromycin-free DMEM served as a cell model of acute Ct infection (acute infection group).After 48-hour infection with Ct,azithromycin was removed,and infected Hela229 cells in the above 2 groups were successively cultured with DMEM for the resurgence of Ct.Immunofluorescence assay and electron microscopy were performed to verify the persistent Ct infection model.The Hela229 cells in the persistent infection group and acute infection group as well as uninfected Hela229 cells (control group) were treated with staurosporine (STS) for 4 hours to induce the apoptosis,and then cell apoptosis was detected by Hoechst 33258 staining,annexin V/propidium iodide staining and flow cytometry.Results After the treatment with azithromycin,atypical inclusions with aberrant reticulate bodies appeared in the Ct-infected cells.After removing azithromycin,cells were cultured until 96 hours after infection,and infectious elementary bodies reappeared in the Ct inclusions.After the treatment with STS,Hoechst staining showed that there was loose chromatin in the persistently infected cells,while chromatin condensation was observed in the uninfected cells.After 24-hour infection with Ct and 4-hour induction with STS,the apoptosis rate was significantly higher in the persistent infection group (45.567％ ± 2.631％) than in the acute infection group (38.567％ ± 1.701％,t =2.686,P =0.028),but significantly lower in the persistent infection group than in the uninfected group (69.800％ ± 2.835％,t =8.187,P ＜ 0.001).After 48-hour infection with Ct and 4-hour induction with STS,there was a significant difference in the apoptosis rate between the persistent infection group (46.700％ ± 5.257％) and acute infection group (61.767％ ± 1.815％,t =5.781,P ＜ 0.001),as well as between the persistent infection group and the uninfected group (68.667％ ± 3.156％,t =7.421,P ＜ 0.001).Conclusion This study showed that azithromycin-induced persistent Ct infection regulated the apoptosis of host cells,and this effect lasted 48 hours.

Objective To analyze the correlation between clinical features and renal dysfunction in patients of acute lacunar infarction with progressive cerebral microbleeds (CMBs). Methods Two hundred and sixty-five patients with first-episode acute lacunar infarction were selected. The serum creatinine was measured within 24 h of admission and the estimated glomerular filtration rate (eGFR) was calculated. The brain MRI (including gradient-echo images) was examined within 2 d of admission and after 1 years of follow-up, respectively. The progressive CMBs was assessed with microbleeds anatomical rating scale (MARS), and the patients were divided into progressive CMBs group (progressive group, 42 cases) and non progressive CMBs group (non progressive group, 223 cases). The clinical features of 2 groups were compared and the correlation between progressive CMBs and renal dysfunction was analyzed. Results The age, 24 h pulse pressure, incidences of renal dysfunction and CMBs in progressive group were significantly higher than those in non progressive group: (69.8 ± 5.8) years vs. (61.5 ± 4.9) years, (63.3 ± 3.1) mmHg (1 mmHg=0.133 kPa) vs. (51.8 ± 4.2) mmHg, 69.0％(29/42) vs. 39.9％(89/223) and 57.1％(24/42) vs. 25.1％(56/223), and the platelet was significantly lower than that in non-progression group:(168 ± 35) ×109/L vs. (189 ± 40) ×109/L, and there were statistical differences (P<0.05 or<0.01). The Logistic regression analysis result showed that renal dysfunction and CMBs were Independent risk factors of progressive CMBs (OR = 1.571 and 1.054, 95％ CI 1.042 - 2.493 and 1.010 - 1.142, P<0.05). Conclusions The rate of renal dysfunction is higher in patients of acute lacunar infarction with progressive CMBs, and progressive CMBs are associated with renal dysfunction.

Objective To investigate the relationship between different topographic locations and neurological deteriorations (ND) in patients with acute new isolated pontine infarction.Methods One hundred sixty-eight patients with acute new isolated pontine infarction during arch 2012 to March 2016 were identified by diffusion weighted imaging (DWI) for retrospective review.Patients were divided into two groups according to their clinical symptoms:patients with ND and patients without ND.According to neuroimaging of DWI,the topographic location of pontine infarction was divided into three types:The upper,middle,and lower ones,and the correlations of ND with risk factors,laboratory examination results,clinical manifestations and different topographic locations were explored by statistical tests.Results Of 168 patients,26.8％ (45/168) were diagnosed with ND,and 73.2％ (123/168) were diagnosed without ND.Univariate analysis showed that there were differences in female ratio [62.2％ (28/45) vs 41.5％ (51/ 123)],smoking ratio [13.3％ (6/45) vs 26.0％ (32/123)],mean length of hospital stay [(22.83 ± 7.12)d vs (19.31 ± 7.65)d],ratio of worse short-term clinical outcomes [77.8％ (35/45) vs 33.3％ (41/123)],and ratio of lower pontine infarction [55.6％ (25/45) vs 26.0％ (32/123)] between two groups (P ＜ 0.05).Logistic regression analysis showed that lower pontine infarction was the independent risk factor of ND (OR =1.953,95％ CI:1.092-3.535,P =0.029).Conclusions Topographic location of lower pons lesions may be reliable predictor of ND in acute new isolated pontine infarction.