Nitrogen availability plays a pivotal role in shaping the composition of root-associated microbial consortia in plants. Nevertheless, elucidating the mechanisms by which nitrogen availability regulates microbial populations and their metabolic activities across different root-associated ecological niches requires further investigation. This research employed an integrative approach combining microbiological approaches with non-targeted metabolomic analyses to examine nitrogen-mediated variations in microbial communities and metabolic processes within ginseng root systems. High-throughput sequencing alongside UPLC-MS/MS analytical platforms was utilized to conduct this multidimensional investigation. Our findings reveal that ginseng treated with N1 exhibited significantly increased yield by 29.90% compared to N0 and by 38.05% compared to N2 (p < 0.05). Additionally, nitrogen application markedly reduced the diversity of microbial communities within various segments of the root system, including rhizosphere soil (RS), rhizoplane soil (TS), fibrous roots (F), and phloem (P). Concurrently, there was a shift in bacterial communities from oligotrophic to eutrophic groups, with specific enrichment of groups such as Rhodanobacter and Burkholderia-Caballeronia-Paraburkholderia, which play crucial roles in the nitrogen cycling process. Metabolomic profiling revealed substantial modifications in soil metabolite profiles under nitrogen treatment, with marked alterations detected across 11 critical biochemical pathways encompassing plant-derived secondary metabolite biosynthesis and environmental microbial metabolic processes. Correlation analysis further indicated that the yield of ginseng and total ginsenoside content in F consistently varied in conjunction with soil nitrate nitrogen (NO₃⁻-N) content in the RS. Additionally, m-cresol was found to play a pivotal role in inhibiting the pathogenic fungus Alternaria, actively responding to pH fluctuations and promoting the synthesis of total ginsenosides in ginseng. These insights elucidate the complex interplay between nitrogen levels and both microbial and metabolomic dynamics, providing a foundational understanding for the strategic manipulation of microbial communities to enhance the sustainability of ginseng agriculture.

Inactivated vaccines based on cell culture are very useful in the prevention and control of many diseases. The most popular strategy for the production of inactivated vaccines is based on monkey-derived Vero cells, which results in high productivity of the virus but has a certain carcinogenic risk due to non-human DNA contamination. Since human diploid cells, such as MRC-5 cells, can produce a safer vaccine, efforts to develop a strategy for inactivated vaccine production using these cells have been investigated using MRC-5 cells. However, most viruses do not replicate efficiently in MRC-5 cells. In this study, we found that rabies virus (RABV) infection activated a robust interferon (IFN)-β response in MRC-5 cells but almost none in Vero cells, suggesting that the IFN response could be a key limiting factor for virus production. Treatment of the MRC-5 cells with IFN inhibitors increased RABV titers by 10-fold. Additionally, the RABV titer yield was improved five-fold when using IFN receptor 1 (IFNAR1) antibodies. As such, we established a stable IFNAR1-deficient MRC-5 cell line (MRC-5^IFNAR1−), which increased RABV production by 6.5-fold compared to normal MRC-5 cells. Furthermore, in a pilot-scale production in 1500 square centimeter spinner flasks, utilization of the MRC-5^IFNAR1− cell line or the addition of IFN inhibitors to MRC cells increased RABV production by 10-fold or four-fold, respectively. Thus, we successfully established a human diploid cell-based pilot scale virus production platform via inhibition of IFN response for rabies vaccines, which could also be used for other inactivated virus vaccine production.

Genital herpes (GH) has become one of the most common sexually transmitted diseases worldwide, and it is spreading rapidly in developing countries. Approximately 90% of GH cases are caused by HSV-2. Therapeutic HSV-2 vaccines are intended for people already infected with HSV-2 with the goal of reducing clinical recurrences and recurrent virus shedding. In our previous work, we evaluated recombinant adenovirus-based vaccines, including rAd-gD2ΔUL25, rAd-ΔUL25, and rAd-gD2, for their potency as prophylactic vaccines. In this study, we evaluated these three vaccines as therapeutic vaccines against acute and recurrent diseases in intravaginal challenged guinea pigs. Compared with the control groups, the recombinant vaccine rAd-gD2ΔUL25 induced a higher titer of the binding antibody, and rAd-gD2 + rAd-ΔUL25 induced a higher titer of the neutralizing antibody. Both rAd-gD2ΔUL25 and rAd-gD2 + rAd-ΔUL25 vaccines significantly enhanced the survival rate by 50% compared to rAd-gD2 and reduced viral replication in the genital tract and recurrent genital skin disease. Our findings provide a new perspective for HSV-2 therapeutic vaccine research and provide a new technique to curtail the increasing spread of HSV-2.

HIV-1 vaccines have been challenging to develop, partly due to the high level of genetic variation in its genome. Thus, a vaccine that can induce cross-reactive neutralization activities will be needed. Studies on the co-evolution of antibodies and viruses indicate that mimicking the natural infection is likely to induce broadly neutralizing antibodies (bnAbs). We generated the consensus Env sequence for each time point in subject CH505, who developed broad neutralization activities, and selected five critical time points before broad neutralization was detected. These consensus sequences were designed to express stable Env trimers. Priming with the transmitted/founder Env timer and sequential boosting with these consensus Env trimers from different time points induced broader and more potent neutralizing activities than the BG505 Env trimer in guinea pigs. Analysis of the neutralization profiles showed that sequential immunization of Env trimers favored nAbs with gp120/gp41 interface specificity while the BG505 Env trimer favored nAbs with V2 specificity. The unique features such as consensus sequences, stable Env trimers and the sequential immunization to mimic natural infection likely has allowed the induction of improved neutralization responses.

Soil, as the foundation for the survival of understory ginseng, directly impacts its growth and development. However, studies focusing on the role of soil in determining the quality of understory ginseng are limited. This study examines the relationship between quality and yield of 5-, 9-, and 17-year-old understory ginseng and their soil microbiota. The results indicate that with the increase in growth years, the overall biomass of understory ginseng generally shows an upward trend, while its quality slightly decreases at 9 years. Compared to the other two growth years, the soil from 9-year-old ginseng shows lower enzyme activity and pH and a higher abundance of pathogens. The 17 years soil has higher OM and AHN content, along with increased abundance of denitrifying and nitrogen-reducing bacteria. Correlation networks reveal that AK significantly influences ginsenoside content, while AP and AHN are more closely related to soil microorganisms. Compared with other types of ginsenosides, ginsenosides Rh2 and Rd are affected by a greater variety of soil microorganisms and chemical factors. As growth years increase, the changes in ginseng quality, soil nutrients, and soil microbiota do not follow a single linear trend; instead, there appears to be a bottleneck phase at certain intermediate stages.