ObjectiveTo study the pharmacological substances and mechanisms through which sesquiterpene ZH-13 from Aquilariae Lignum Resinatum improves neuroinflammation. MethodsBV-2 microglial cells were stimulated with lipopolysaccharide （LPS） to induce neuroinflammation. The cells were divided into the normal group， the model group， and the ZH-13 low- and high-dose treatment groups （10， 20 μmol·L^-1）. The model group was treated with 1 μmol·L^-1 LPS. Cell viability was assessed using the cell proliferation and activity assay （CCK-8 kit）. Nitric oxide （NO） release in the cell supernatant was measured using a nitric oxide kit （Griess method）. The mRNA expression levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， inducible nitric oxide synthase （iNOS）， and interleukin-6 （IL-6） were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The phosphorylation of mitogen-activated protein kinase （MAPK） pathway proteins was assessed by Western blot. ResultsCompared with the model group， ZH-13 dose-dependently reduced NO release from BV-2 cells under LPS stimulation （P<0.05， P<0.01）. In the 20 μmol·L^-1 ZH-13 treatment group， the mRNA expression levels of IL-1β， TNF-α， iNOS， and IL-6 were significantly reduced compared to the model group （P<0.05， P<0.01）. In both the low- and high-dose ZH-13 groups， the expression of the inflammatory factor TNF-α and the phosphorylation of c-Jun N-terminal kinase （JNK） in the upstream MAPK pathway were significantly reduced （P<0.05）. After stimulation with the JNK agonist anisomycin （Ani）， both low- and high-dose ZH-13 treatment groups showed reduced phosphorylation of JNK proteins compared to the Ani-treated group （P<0.01）. ConclusionThe sesquiterpene compound ZH-13 from Aquilariae Lignum Resinatum significantly ameliorates LPS-induced neuroinflammatory responses in BV-2 cells by inhibiting excessive JNK phosphorylation and reducing TNF-α expression. These findings elucidate the pharmacological substances and mechanisms underlying the sedative and calming effects of Aquilariae Lignum Resinatum.

Synthetic biology is a crucial tool for the development of the bio-industry and bio-economy, representing a significant aspect of new quality productive forces. As a core course for graduate students in bioengineering, Synthetic Biology plays a vital role in ensuring the supply of essential talents for the development of the bio-industry in the new era. To better serve regional economic development and provide high-level talents for China's progress in the bio-industry, we analyzed typical issues encountered in the past teaching activities, set up a multi-disciplinary teaching team, optimized the course contents, adjusted the teaching mode, and mobilized students' learning interest. With the application of scientific research project as the starting point, we guided students to think and discuss deeply through the simulation of application writing and project defense, which improved students' critical thinking and innovative thinking. With industrialization as a focus, we explored a new training model combining production, education, and research through the joint practice base of the university and enterprises introduced typical cases of biomanufacturing to encourage students to engage in scientific research. The teaching reform significantly enhances the comprehensive abilities and national sentiments of graduate students. This paper hopes to serve as a reference for colleagues engaged in teaching in this field.
Synthetic Biology/education* ; Teaching ; China ; Humans

This study investigates the effect of glycyrrhetinic acid(GA) combined with doxorubicin(DOX) on apoptosis in HepG2 cells and its possible mechanisms. HepG2 cells were cultured in vitro, and cell viability was assessed using the cell counting kit-8(CCK-8) method. Flow cytometry was used to measure apoptosis levels in HepG2 cells. The cells were divided into the following groups: control group(0 μmol·L~(-1)), DOX group(2 μmol·L~(-1)), GA group(150 μmol·L~(-1)), and DOX + GA combination group(2 μmol·L~(-1) DOX + 150 μmol·L~(-1) GA), with treatments given for 24 hours. The colocalization level between the endoplasmic reticulum(ER) and mitochondria was assessed by colocalization fluorescence imaging. Fluorescence probes were used to measure the Ca~(2+) content in the ER and mitochondria. The qRT-PCR and Western blot were used to determine the mRNA and protein expression of sirtuin-3(SIRT3). Co-immunoprecipitation(CO-IP) was applied to investigate the interactions between voltage-dependent anion channel 1(VDAC1) and SIRT3, as well as between VDAC1, glucose-regulated protein 75(GRP75), and inositol 1,4,5-trisphosphate receptor(IP3R). The results showed that the combination of DOX and GA promoted apoptosis in HepG2 liver cancer cells. The colocalization level between the ER and mitochondria was significantly reduced, the Ca~(2+) content in the ER was significantly increased, and the Ca~(2+) content in the mitochondria was significantly decreased. The relative expression of VDAC1, GRP75, and IP3R was significantly reduced, and interactions between VDAC1, GRP75, and IP3R were observed. SIRT3 mRNA and protein expression levels were significantly increased, and an interaction between SIRT3 and VDAC1 was detected. The acetylation level of VDAC1 was significantly decreased. In conclusion, GA combined with DOX induces apoptosis in HepG2 cells by mediating the deacetylation of VDAC1 through SIRT3, weakening the interactions among VDAC1, GRP75, and IP3R. This regulates the formation of endoplasmic reticulum-mitochondrial membrane domains(ERMMDs), affects Ca~(2+) transport between the ER and mitochondria, and ultimately triggers cell apoptosis.
Humans ; Apoptosis/drug effects* ; Hep G2 Cells ; Glycyrrhetinic Acid/pharmacology* ; Doxorubicin/pharmacology* ; Liver Neoplasms/genetics* ; Carcinoma, Hepatocellular/physiopathology* ; Mitochondria/metabolism* ; Endoplasmic Reticulum/metabolism* ; Cell Survival/drug effects* ; Membrane Proteins/genetics*

Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

Jansen-de Vries syndrome, also known as intellectual developmental disorder with gastrointestinal difficulties and high pain threshold, is a rare autosomal dominant disorder characterized by multisystem involvement. This article reports the case of a young child who presented with global developmental delay, gastrointestinal dysfunction, intellectual disability, and short stature. Distinct facial features included a broad forehead, low nasal bridge, thin upper lip, and widely spaced and misaligned teeth. Additional phenotypic findings involved small hands and feet, as well as digital anomalies. Through whole-exome sequencing (WES) and copy number variation (CNV) analysis, a pathogenic variant was identified in the PPM1D gene: c.1281G > A (p.Trp427Ter). This report also reviews the current literature on the clinical characteristics, diagnostic approaches, and therapeutic advances related to this condition, aiming to provide valuable insights for its clinical diagnosis and management.

Objective To explore the underlying mechanisms of ulcerative colitis(UC)pathogenesis using single-cell transcriptomic bioinformatics approaches.Methods Single-cell transcriptome dataset GSE125527,consisting of UC samples,was downloaded from a high-throughput Gene Expression Omnibus for this study.Data filtering and normalization were conducted using the Seurat package in R.Cells with similar gene expression profiles were clustered and annotated.Cell subgroups associated with UC pathogenesis were identified,and differ-entially expressed genes(DEGs)were extracted.Enrichment analysis of these DEGs was performed to uncover potential signaling pathways involved in UC.High-dimensional weighted gene co-expression network analysis(hdW-GCNA)was applied to the cell subgroup with the greatest differences to select UC-related gene modules.Additional datasets,GSE36807,GSE42911,GSE65114,and GSE6731,were downloaded for DEG screening.By integrating single-cell DEGs with UC-related gene modules,core genes involved in UC pathogenesis were identified.Receiver operating characteristic(ROC)curves were plotted to evaluate the predictive value of these core genes for UC pathogenesis.The expression of core genes was validated in a UC rat model.Results After filtering,18642 cells from UC samples and 4298 cells from normal colonic tissue were obtained and clustered into ten subgroups,with a high proportion of B cells in UC samples.Eighty-one upregulated and twenty-one downregulated DEGs related to mononuclear cells were identified.Significant enrichment of DEGs was observed in pathways related to tumor necro-sis factor-alpha,nuclear factor Kappa B,interleukin-2,signal transducer and activator of transcription 5,and interferon-alpha.Two UC-related gene modules were identified through hdWGCNA.After screening and validation,LAPTM5 was confirmed as a core gene in UC pathogenesis.Compared to control rats,LAPTM5 expression was significantly higher in the colonic tissues of UC model rats.Conclusions There are significant differences in cell distribution between UC and normal colonic tissues,with B cells closely related to UC pathogenesis,serving as potential targets for prevention and treatment.LAPTM5 is identified as a potential core gene in UC pathogenesis.

Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.

Objective To investigate the inhibitory effect and mechanism of lentinan on colitis-associated colorectal cancer (CAC) induced by azomethane (AOM)/dextran sulfate sodium salt (DSS) through the IL-6/ STAT3 pathway. Methods C57BL/6 mice were randomly divided into a control group, a model group, a low-dose group (0.865 mg/kg lentinan), a medium-dose group (1.73 mg/kg lentinan), and a high-dose group (3.46 mg/kg lentinan). Except the control group, CAC was induced by AOM/DSS in the other groups, and corresponding drugs were injected intraperitoneally during the modeling process. Body mass, disease activity index (DAI) score, colon length, and tumor number were compared among all groups. Hematoxylin–eosin staining was used to observe the pathological morphology of colon. ELISA was utilized to detect the IL-6, IL-1β, and IL-18 contents in serum. Western blot analysis was conducted to detect the expression levels of IL-6, p-STAT3, and c-Myc in colon tissues. Results The tumor number, DAI score, serum IL-6, IL-1β, and IL-18 contents and the expression levels of IL-6, p-STAT3, and c-Myc in the colon tissue of the model group were higher than those of the control group, while the body mass and colon length were lower than those of the control group (P<0.05). The pathological morphology of colon tissues showed adenocarcinoma formation. After different doses of lentinan intervention, the tumor number, DAI score, serum IL-6, IL-1β, and IL-18 contents and the expression levels of IL-6, p-STAT3, and c-Myc in colon tissues were all lower than those in the model group, while body mass and colon length were higher than those in the model group (P<0.05). The pathological morphology of colon tissues showed adenomas of different grades but no adenocarcinoma was found. Conclusion Lentinan inhibits CAC formation, and its anticancer effect is related to the inhibition of the IL-6/STAT3 pathway.

Bacterial resistance has always been a challenge in the field of antibacterials research. As one of the most problematic pathogens causing nosocomial infections, Acinetobacter baumannii (A. baumannii) was a severe threaten to public health. In recent years, as the rate of multidrug-resistant, extensively drug-resistant, and even pan drug-resistant A. baumannii has been increasing, there is an urgent need to develop new strategies to combat this pathogen. Iron is an essential element for the survival and the infection process of A. baumannii. Bacteria have developed a series of iron uptake strategies to survive in conditions of the host iron starvation, including the secretion of siderophore for iron chelation, the uptake of ferroheme, and the transport of ferrous by Feo system. Developing antibacterial strategies to inhibit the iron uptake is an effective way to deal with A. baumannii infection. This article will review the mechanisms of iron uptake by A. baumannii and the antimicrobial strategies developed based on them.

BACKGROUND:As a common complication of diabetes mellitus,male reproductive disorders have received increasing attention in recent years.Ganoderma spore have hypoglycemic,antioxidant and anti-inflammatory effects,but the regulatory mechanism for diabetic testicular tissue has not been fully elucidated. OBJECTIVE:To investigate the effect of ganoderma spore on the PTEN-induced kinase 1/E3 ubiquitin ligase pathway and cell apoptosis in testicular tissue of diabetic rats. METHODS:Forty male Sprague-Dawley rats were randomly divided into normal group,high fat and high sugar group,diabetic group and ganoderma spore group,with 10 rats in each group.The latter three groups were given high fat/high sugar diet until the end of the experiment.After 1 month of high fat/high sugar diet,the diabetic and ganoderma spore groups were given intraperitoneal injection of streptozotocin(30 mg/kg per day)to establish type 2 diabetic rat models.After successful modeling,the ganoderma spore group was intragastrically given ganoderma spore(300 mg/kg per day),and the other groups were given the same amount of normal saline for continuous 12 weeks.The sperm number and morphology were detected.The histopathological changes of the testicle were observed.Serum testosterone and oxidative stress levels in testicular tissue were measured.The levels of PTEN-induced kinase 1,E3 ubiquitin ligase,and anti-nucleoporin p62 were analyzed by immunohistochemistry and the expression of PTEN-induced kinase 1,E3 ubiquitin ligase,anti-nucleoporin p62,programmed cell death-1,microtubule-associated protein light chain 3 Ⅱ/Ⅰ,caspase 3,cleaved-caspase 3 were detected by western blot assay. RESULTS AND CONCLUSION:Compared with the normal group and the high fat and high sugar group,the diabetic group had decreased sperm number(P＜0.01),increased sperm malformation rate(P＜0.01),and decreased serum testosterone level(P＜0.01).Compared with the diabetic group,ganoderma spore intervention could increase the sperm number(P＜0.05),decrease the malformation rate(P＜0.01),and increase the serum testosterone level(P＜0.01).Compared with the normal group and the high fat and high sugar group,the malondialdehyde level in testis tissue was increased in the diabetic group(P＜0.01),while the levels of glutathione deoxidase and superoxide dismutase decreased(P＜0.01).Compared with the diabetic group,the malondialdehyde level in testis tissue was decreased in the ganoderma spore group(P＜0.01),and the levels of glutathione deoxidase and superoxide dismutase increased(P＜0.01).Immunohistochemical staining showed that compared with the normal group and the high fat and high sugar group,the positive expressions of PTEN-induced kinase 1 and E3 ubiquitin ligase in testicular tissue were decreased in the diabetic group,while the positive expressions of anti-nucleoporin p62 were increased.Compared with the diabetic group,the positive expressions of PTEN-induced kinase 1 and E3 ubiquitin ligase in testicular tissue e were increased in the ganoderma spore group,while the positive expression of anti-nucleoporin p62 was decreased.Western blot assay results indicated that compared to the normal group and the high fat and high sugar group,the expression of PTEN-induced kinase 1 and E3 ubiquitin ligase,programmed cell death-1 and the ratio of microtubule-associated protein light chain 3 Ⅱ/Ⅰ protein were decreased in the diabetic group(P＜0.05 or P＜0.01),while the expressions of anti-nucleoporin p62,caspase3 and cleaved-caspase3 were increased(P＜0.01).Compared with the diabetic group,ganoderma spore intervention could elevate the expression of PTEN-induced kinase 1 and E3 ubiquitin ligase,programmed cell death-1 and the ratio of microtubule-associated protein light chain 3 Ⅱ/Ⅰ protein(P＜0.05 or P＜0.01)as well as reduce the expressions of anti-nucleoporin p62,caspase3 and cleaved-caspase3(P＜0.05 or P＜0.01).Overall,ganoderma spores may activate the PTEN-induced kinase 1/E3 ubiquitin ligase pathway to enhance autophagy in testicular tissue and reduce apoptosis in tissue cells,so as to protect testicular tissue.