Effect of LRRC15 on autophagy in A549 cells.

Idiopathic pulmonary fibrosis (IPF) is a progressive, chronic, and irreversible interstitial lung disease with unknown cause. To explore the role and regulatory mechanism of leucine-rich repeat-containing protein 15 (LRRC15) in IPF, bleomycin (BLM)-induced pulmonary fibrosis in mouse and A549 cells were constructed, and the expression of LRRC15 were detected. Then, MTT, GFP-RFP-LC3 dual fluorescent labeling system and Western blotting were used to investigate the effects of LRRC15 on cell activity and autophagy after transfection of siLRRC15, respectively. The results indicated that the expression of LRRC15 was significantly increased after the BLM treatment in mouse lung tissue and A549 cells. The designed and synthesized siLRRC15 followed by transfection into A549 cells resulted in a dramatic reduction in LRRC15 expression and partially restored the cell damage induced by BLM. Moreover, the expression of LC3-II and P62 were up-regulated, the amount of autophagosome were increased by GFP-RFP-LC3 dual fluorescent labeling assay after BLM treatment. Meanwhile, this study also showed that the key autophagy proteins LC3-II, ATG5 and ATG7 were up-regulated, P62 was down-regulated and autophagic flux were enhanced after further treatment of A549 cells with siLRRC15. The above findings suggest that LRRC15 is an indicator of epithelial cell damage and may participate in the regulation of fibrosis through autophagy mechanism in IPF. This study provides necessary theoretical basis for further elucidating the mechanism of IPF.

Pancreas-directed AAV8-hSPINK1 gene therapy safely and effectively protects against pancreatitis in mice.

OBJECTIVE: Currently, there is no cure for chronic pancreatitis (CP). Germline loss-of-function variants in SPINK1 (encoding trypsin inhibitor) are common in patients with CP and are associated with acute attacks and progression of the disease. This preclinical study was conducted to explore the potential of adeno-associated virus type 8 (AAV8)-mediated overexpression of human SPINK1 (hSPINK1) for pancreatitis therapy in mice. DESIGN: A capsid-optimised AAV8-mediated hSPINK1 expression vector (AAV8-hSPINK1) to target the pancreas was constructed. Mice were treated with AAV8-hSPINK1 by intraperitoneal injection. Pancreatic transduction efficiency and safety of AAV8-hSPINK1 were dynamically evaluated in infected mice. The effectiveness of AAV8-hSPINK1 on pancreatitis prevention and treatment was studied in three mouse models (caerulein-induced pancreatitis, pancreatic duct ligation and Spink1 c.194+2T>C mouse models). RESULTS: The constructed AAV8-hSPINK1 vector specifically and safely targeted the pancreas, had low organ tropism for the heart, lungs, spleen, liver and kidneys and had a high transduction efficiency (the optimal expression dose was 2×1011 vg/animal). The expression and efficacy of hSPINK1 peaked at 4 weeks after injection and remained at significant level for up to at least 8 weeks. In all three mouse models, a single dose of AAV8-hSPINK1 before disease onset significantly alleviated the severity of pancreatitis, reduced the progression of fibrosis, decreased the levels of apoptosis and autophagy in the pancreas and accelerated the pancreatitis recovery process. CONCLUSION: One-time injection of AAV8-hSPINK1 safely targets the pancreas with high transduction efficiency and effectively ameliorates pancreatitis phenotypes in mice. This approach is promising for the prevention and treatment of CP.

Relationship between parents attitudes towards sexual education and their proactive adolescent sexual education for primary and secondary students in Baoshan City of Yunnan Province / 中国学校卫生

Objective@#To understand parents attitudes towards adolescent sexual education in Baoshan City of Yunnan Province, and to explore the correlation between attitudes and their proactive behavior in adolescent sexual education, so as to provide reference for improving parents initiative in adolescent sexual health education.@*Methods@#From November to December 2022, a stratified random cluster sampling method was used to select 7 389 parents of students from 35 primary, middle, and high schools in Baoshan City of Yunnan Province for a questionnaire survey on adolescent sexual health education in parents schools. Chi-square tests were conducted to compare the attitudes of parents towards sexual health education among different groups and their proactive behavior in providing sexual health education to their children. A multivariate Logistic regression model was used to analyze the correlation between parents attitudes towards sexual education and proactive sexual health education.@*Results@#Totally 43.17% ( 3 190 ) of parents had a more open attitude towards sex education for their children, with parents of girls, parents of elementary school students, mothers, <30 years old, junior high school education, workers, service personnel, married, harmonious relationships with their children and democratic parents had a higher rate of actively providing sex education for their children ( χ 2=4.65, 401.92, 23.53 , 197.50, 38.18, 13.65, 6.54, 8.07, 32.17, 96.58, P <0.05). Parents with a more conservative attitude towards sex education and parents of boys were negatively correlated with parents actively providing adolescent sexual health education to their children ( OR =0.86, 0.38, P <0.05). Parents who were <30, 30-<40, 40-50 years old, individuals/businesses, married, divorced, had a good and decent relationship with their children were positively correlated with their parents proactive adolescent sexual health education for their children ( OR =1.50, 1.90, 1.37, 1.22, 1.60, 1.32, 1.94, 1.53, P <0.05).@*Conclusion@#A more open attitude towards adolescent sexual health education can serve as a predictive factor for proactive adolescent sexual health education. In order to increase the rate of family sexual health education, it is recommended to enhance parents awareness of their responsibility for their children s sexual health education, promote harmonious parent-child relationships between parents and children, and communicate with their children with an open attitude.