Comparison of bacterial diversity in Bactrocera cucurbitae (Coquillett) ovaries and eggs based on 16S rRNA sequencing.

Next-generation sequencing allows for fine-scale studies of microbial communities. Herein, 16S ribosomal RNA high-throughput sequencing was used to identify, classify, and predict the functions of the bacterial communities in the eggs and ovaries of Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), which is a pest that infests a variety of cucurbit fruits at different developmental stages. Taxonomic analyses indicate that bacteria associated with B. cucurbitae represent 19 phyla, which were spread across different developmental stages. Specifically, the egg microbiota had a higher alpha diversity than those of microbiota in the primary and mature ovaries. Significant differences were not observed between the primary and mature ovaries in terms of their microbiota's alpha diversities. Pseudomonadota, Deinococcota, Bacteroidota, Bacillota, and Actinomycetota were the dominant phyla in all three developmental stages of B. cucurbitae, and Pseudomonadaceae and Enterobacteriaceae were the most abundant families. Owing to the unique physiological environment of the ovaries, the diversity of their bacterial community was significantly lower than that in the eggs. This study provides new insights into the structure and abundance of the microbiota in B. cucurbitae at different developmental stages and contributes to forming management strategies for this pest.

Effect and Mechanism of Berberine Hydrochloride Against Lung Cancer Cells in Vitro Based on Mevalonate Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the efficacy and mechanism of berberine hydrochloride （BBH） against lung cancer cells through the mevalonate （MVA） pathway. MethodHuman lung cancer A549 cells and mouse Lewis lung carcinoma （LLC） cells were used as research subjects. Cell proliferation and cell counting kit-8 （CCK-8） assay were performed to detect the inhibitory effect of BBH （10， 20， 30， 40， 50 μmol·L-1） on the proliferation of the two kinds of cells （48 h）. Then cell scratch assay was used to explore the influence of BBH （40 μmol·L-1） on the migration of A549 and LLC cells （24， 48 h）， and colony formation assay was conducted to compare the colony formation ability of the cells under different concentrations of BBH （10， 20， 40 μmol·L-1）. Moreover， the effects of BBH （40 μmol·L-1） on the content of acetyl-coenzyme A （A-CoA） and total cholesterol （TC） in A549 and LLC cells were determined by kit assay. AutoDock Vina was used for the dock of BBH and MVA pathway regulatory protein， sterol regulatory element-binding protein 2 （SREBP2）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to observe the effects of BBH （40 μmol·L-1） on the mRNA expression of nine genes related to the MVA pathway in A549 and LLC cells： hydroxymethylglutaryl-CoA synthase 1 （HMGCS1）， hydroxymethylglutaryl-CoA Reductase （HMGCR）， mevalonate kinase （MVK）， phosphomevalonate kinase （PMVK）， mevalonate 5-pyrophosphate decarboxylase （MVD）， farnesyl diphosphate synthase （FDPS）， squalene epoxidase （SQLE）， farnesyl-diphosphate farnesyltransferase 1 （FDFT1）， and geranylgeranyl diphosphate synthase 1 （GGPS1）. Western blot was performed to clarify the effects of BBH （40 μmol·L-1） on the expression of three key proteins of the MVA pathway： HMGCS1， HMGCR， and FDFT1. The Cancer Genome Atlas （TCGA） database was searched to analyze the relationship between HMGCS1， HMGCR， FDFT1 and transcription gene SREBF2 in non-small cell lung cancer （NSCLC）. ResultCompared with the conditions in the control group， the proliferation， migration， and colony formation of A549 and LLC cells in the BBH group were decreased （P<0.01）， while the cell apoptosis rate was increased （P<0.01）. Molecular docking showed that BBH had good binding activity with SREBP2. In addition， the content of A-CoA and TC of the MVA pathway was reduced （P<0.01）. BBH down-regulated the mRNA expression of HMGCS1， HMGCR， MVK， PMVK， MVD， FDPS， SQLE， FDFT1， and GGPS1 in A549 and LLC cells （P<0.01）， and lowered the levels of HMGCS1， HMGCR， and FDFT1 proteins （P<0.05， P<0.01）. In NSCLC patients， HMGCS1， HMGCR， and FDFT1 were highly correlated with SREBF2 （R=0.54， R=0.57， and R=0.48）. ConclusionBBH can inhibit the proliferation， migration， and colony formation of A549 and LLC cells and promote cell apoptosis， which may be related to the regulation of MVA pathway by BBH binding to SREBP2.