Selective autophagy keeps cell homeostasis by degrading aggregated proteins, damaged or over-abundant organelles, and other cytoplasmic substances. The maintenance of its normal function needs to ensure that the autophagy receptor can effectively recognize and isolate undegraded substances. As an important autophagy receptor protein, p62 participates in the process of selective autophagy by mediating multiple signaling pathways. Fibrosis is a pathological feature of most chronic inflammatory diseases. When fibrosis develops for a long time, it will cause substantial scar formation and eventually lead to cell dysfunction and organ failure. The accumulation, overexpression and ectopic expression of p62 can aggravate the occurrence and development of lung, liver and kidney fibrosis diseases. Therefore, it is very critical to explore the effect of selective autophagy receptor p62 on fibrotic diseases.

Selective autophagy keeps cell homeostasis by degrading aggregated proteins, damaged or over-abundant organelles, and other cytoplasmic substances. The maintenance of its normal function needs to ensure that the autophagy receptor can effectively recognize and isolate undegraded substances. As an important autophagy receptor protein, p62 participates in the process of selective autophagy by mediating multiple signaling pathways. Fibrosis is a pathological feature of most chronic inflammatory diseases. When fibrosis develops for a long time, it will cause substantial scar formation and eventually lead to cell dysfunction and organ failure. The accumulation, overexpression and ectopic expression of p62 can aggravate the occurrence and development of lung, liver and kidney fibrosis diseases. Therefore, it is very critical to explore the effect of selective autophagy receptor p62 on fibrotic diseases.

Objective:To investigate the effect of smoking on autophagy in alveolar macrophages (AMs) of silicosis patients.Methods:In December 2019, a random sampling method was used to select 42 male patients with silicosis (19 cases of stage II and 23 cases of stage III) who were treated with large volume whole lung lavage from August to December 2017 in the Beidaihe sanatorium. According to the different smoking index of the study subjects (smoking index=smoking cigarette consumptions per day×years of smoking) , we divided them into high (Smoking index>400) , medium (200≤smoking index≤400) , low (smoking index <200) and non-smoking group. The levels of autophagy related proteins LC3, Beclin1, p62 and apoptosis related protein Cleaved Caspase-3 were detected by Western blot. The effects of smoking on autophagy activity of AMs in silicosis were analyzed.Results:The ratio of autophagy related protein LC3 II/LC3 I, the expression of Beclin1, p62, and apoptosis related protein Cleaved Caspase-3 in the high smoking group were significantly higher than that of the middle, low smoking group and the non-smoking group ( P<0.05) . Conclusion:Smoking can aggravate the dysfunction of autophagic degradation in silicosis patients' AMs, which may accelerate the progress of silicosis through increasing apoptosis in AMs.

Objective:To explore the effect of atractylenolide-1 (ATL-Ⅰ) on alveolar macrophages in silicosis patients.Methods:In December 2019, 12 male silicosis patients treated in Beidaihe Sanatorium for Chinese Coal Miners from July to September 2019 were selected by random sampling. Their alveolar macrophages were collected and divided into control group, ATL-Ⅰ group (100 μmol/L) and dimethyl sulfoxide (DMSO) group (100 μmol/L) . The exprossion levels of inflammatory factor interleukin-1β (IL-1β) , interleukin-6 (IL-6) , tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay. The expression levels of autophagy associated protein microtubule associated protein light chain 3 (LC3) , autophagy substrate protein p62, lysosome associated membrane protein 2 (LAMP2) , apoptosis associated protein Cleaved caspase-3, nuclear factor kappa B (NF-κB) and its phosphorylated form (p-NF-κB) were detected by Western blot.Results:Compared with the control group and DMSO group, the expression levels of IL-1β, IL-6, TNF-α in alveolar macrophages decreased significantly in the ATL-Ⅰ group ( P<0.05) , and the expression levels of p-NF-κB, the ratio of LC3-Ⅱ/LC3-Ⅰ also decreased significantly in the ATL-Ⅰ group ( P<0.05) . However, the expression levels of NF-κB, LAMP2, p62 and Cleaved caspase-3 in the ATL-Ⅰ group were not statistically different from those in the control group and DMSO group ( P>0.05) . There was no statistically significant differences in the expression of the above indexes between the control group and DMSO group ( P>0.05) . Conclusion:ATL-Ⅰ may reduce the release of inflammatory factors from alveolar macrophages and inhibit the activity of autophagy in silicosis patients, but it may not reduce the level of apoptosis.

Objective:To investigate the effect of smoking on autophagy in alveolar macrophages (AMs) of silicosis patients.Methods:In December 2019, a random sampling method was used to select 42 male patients with silicosis (19 cases of stage II and 23 cases of stage III) who were treated with large volume whole lung lavage from August to December 2017 in the Beidaihe sanatorium. According to the different smoking index of the study subjects (smoking index=smoking cigarette consumptions per day×years of smoking) , we divided them into high (Smoking index>400) , medium (200≤smoking index≤400) , low (smoking index <200) and non-smoking group. The levels of autophagy related proteins LC3, Beclin1, p62 and apoptosis related protein Cleaved Caspase-3 were detected by Western blot. The effects of smoking on autophagy activity of AMs in silicosis were analyzed.Results:The ratio of autophagy related protein LC3 II/LC3 I, the expression of Beclin1, p62, and apoptosis related protein Cleaved Caspase-3 in the high smoking group were significantly higher than that of the middle, low smoking group and the non-smoking group ( P<0.05) . Conclusion:Smoking can aggravate the dysfunction of autophagic degradation in silicosis patients' AMs, which may accelerate the progress of silicosis through increasing apoptosis in AMs.

Objective:To explore the effect of atractylenolide-1 (ATL-Ⅰ) on alveolar macrophages in silicosis patients.Methods:In December 2019, 12 male silicosis patients treated in Beidaihe Sanatorium for Chinese Coal Miners from July to September 2019 were selected by random sampling. Their alveolar macrophages were collected and divided into control group, ATL-Ⅰ group (100 μmol/L) and dimethyl sulfoxide (DMSO) group (100 μmol/L) . The exprossion levels of inflammatory factor interleukin-1β (IL-1β) , interleukin-6 (IL-6) , tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay. The expression levels of autophagy associated protein microtubule associated protein light chain 3 (LC3) , autophagy substrate protein p62, lysosome associated membrane protein 2 (LAMP2) , apoptosis associated protein Cleaved caspase-3, nuclear factor kappa B (NF-κB) and its phosphorylated form (p-NF-κB) were detected by Western blot.Results:Compared with the control group and DMSO group, the expression levels of IL-1β, IL-6, TNF-α in alveolar macrophages decreased significantly in the ATL-Ⅰ group ( P<0.05) , and the expression levels of p-NF-κB, the ratio of LC3-Ⅱ/LC3-Ⅰ also decreased significantly in the ATL-Ⅰ group ( P<0.05) . However, the expression levels of NF-κB, LAMP2, p62 and Cleaved caspase-3 in the ATL-Ⅰ group were not statistically different from those in the control group and DMSO group ( P>0.05) . There was no statistically significant differences in the expression of the above indexes between the control group and DMSO group ( P>0.05) . Conclusion:ATL-Ⅰ may reduce the release of inflammatory factors from alveolar macrophages and inhibit the activity of autophagy in silicosis patients, but it may not reduce the level of apoptosis.

ObjectiveTo explore the association between triglyceride-glucose （TyG） index and major adverse cardiovascular events （MACEs） risk in coronary heart disease （CHD） patients with blood stasis syndrome after percutaneous coronary intervention （PCI）. MethodsA total of 857 CHD patients with blood stasis syndrome after PCI were enrolled and divided into four groups according to the baseline TyG index quartiles， Q1 （TyG ＜ 8.51）， Q2 （8.51 ≤ TyG ＜ 8.88）， Q3 （8.88 ≤ TyG ＜ 9.22）， and Q4 （TyG ≥ 9.22）. The clinical outcome was defined as a compound endpoint of cardiovascular events including cardiac death， non-fatal myocardial infarction， unplanned revascularization， in-stent restenosis and stroke. The machine learning Boruta algorithm was used for feature selection related to MACEs risk. Kaplan-Meier survival analysis and Cox proportional hazards regression model were used to compare the differences in MACEs risk among the four groups. Restricted cubic spline （RCS） and subgroup analysis were performed to determine the relationship between the TyG index and MACEs risk. The area under the receiver operating characteristic （ROC） curve （AUC）， calibration curve and Hosmer-Lemeshow test， and decision curve analysis （DCA） were plotted to evaluate the predictive value of the TyG index for MACEs risk. ResultsThe median follow-up time of the included patients was 2.45 years. During the follow-up period， 313 cases （36.52%） of new MACEs occurred. The incidence of MACEs in Q1， Q2， Q3， Q4 group was 28.17% （60/213）， 29.05% （61/210）， 39.45% （86/218） and 49.07% （106/216）， respectively. Kaplan-Meier survival analysis suggested statistically significant differences in MACEs risk among the four groups （P＜0.001）. Cox proportional hazards regression model analysis found that the risk of MACEs in patients with high TyG index increased by 60.1% （P＜0.01）. Using Q1 as the reference， the MACEs risk in Q2， Q3 and Q4 groups gradually increased， and the trend was statistically significant （P＜0.05）. RCS model suggested that the TyG index was nonlinearly associated with the MACEs risk （P＜0.001）. The TyG index had a good predictive performance for MACEs risk according to ROC analysis （AUC=0.758， 0.724-0.792） and Hosmer-Lemeshow test （χ² = 4.319， P = 0.827）. Additionally， DCA analysis also suggested a good clinical efficacy of the TyG index for predicting MACEs. Subgroup analysis showed that different baseline TyG index was positively correlated with the MACEs risk in the stratification of age， male， BMI， history of diabetes and hypertension， and low-density lipoprotein cholesterol （LDL-C）≥1.8 mmol·L^-1（P＜0.05）. ConclusionThe elevated TyG index is closely corrected with an increased risk of MACEs in CHD patients with blood stasis syndrome after PCI， indicating a guiding significance for early risk stratification and clinical decisions.

Objective:To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province.Methods:A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti -HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results:There were 104 children in the case group and 159 in the control group. The median ( Q1, Q3) level of anti -HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group ( P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 ( OR=1.18, 95% CI: 1.03-1.35) and 93 ( OR=1.21, 95% CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion:The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.

Objective:To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province.Methods:A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti -HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results:There were 104 children in the case group and 159 in the control group. The median ( Q1, Q3) level of anti -HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group ( P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 ( OR=1.18, 95% CI: 1.03-1.35) and 93 ( OR=1.21, 95% CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion:The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.

ObjectiveTo explore the underlying mechanism of Bushen Huatan prescription in alleviating postmenopausal osteoporosis （PMOP） by maintaining the balance of osteogenesis and adipogenic differentiation in ovariectomized rats with osteoporosis. MethodSeventy-five 6-month-old non-pregnant female SD rats were randomly divided into sham-operation group， model group， atorvastatin group， liviol group， and Bushen Huatan prescription group. Bilateral ovaries were removed in the four groups except the sham-operation group， while only the same mass of adipose tissue around the ovaries was removed in the sham-operation group. On the 5^th week after surgery， drugs were consecutively administrated for 8 weeks. Rats in the Bushen Huatan prescription group received 9.4 mg·kg^-1 of the prescription， rats in the atorvastatin group received 0.92 mg·kg^-1 of atorvastatin， rats in the Liviol group received 0.23 mg·kg^-1 of liviol， and rats in the model group and the sham-operation group received saline once a day. Micro-computed tomography （Micro CT） was used to detect bone mineral density （BMD） of rat tibia in each group. Hematoxylin-eosin （HE） staining was used to detect the relative area of rat bone marrow adipose tissue （BMAT） in each group. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the relative expression levels of Runt-related transcription factor 2 （Runx2）， peroxisome proliferator-activated receptor （PPARγ）， leptin （LPN）， and leptin receptor （OBR） in bone tissues. ResultAs compared with the sham operation group， the BMD of rats in the model group decreased （P<0.05）， while the relative area of BMAT increased （P<0.05）. In addition， the expression levels of LPN， OBR， and Runx2 decreased in the model group （P<0.05）， while the level of PPARγ increased （P<0.05）. As compared with the model group， the BMD of rats in the atorvastatin group， the Livial group， and the Bushen Huatan prescription group increased （P<0.05）， and the relative area of BMAT decreased （P<0.05）. The expression levels of LPN， OBR， and Runx2 in these groups increased （P<0.05）， while the expression level of PPARγ decreased （P<0.05）. ConclusionBushen Huatan prescription plays the anti-osteoporosis role in the rat model of PMOP through up-regulating LPN and OBR in bone tissues and maintaining the balance of osteogenesis and adipogenic differentiation， thereby reducing postmenopausal bone loss and playing a role in the prevention and treatment of PMOP.