Effects of the tumor necrosis factor on hemocyte proliferation and bacterial infection in Chinese mitten crab (Eriocheir sinensis).

Tumor necrosis factor (TNF) is an important cytokine that can regulate a variety of cellular responses by binding tumor necrosis factor receptor (TNFR). We studied whether the TNF of Eriocheir sinensis can regulate hemocyte proliferation. The results showed that the EsTNF and EsTNFR were constitutively expressed in all tested tissues, including the heart, hepatopancreas, muscles, gills, stomachs, intestines, and hemocytes. We found that low levels of EsTNF and EsTNFR transcripts were present in hemocytes. The gene expression levels were significantly increased in the hemocytes after being stimulated by Staphylococcus aureus or Vibrio parahaemolyticus. We also found some genes related to cell proliferation were expressed at a higher level in pulsing rTNF-stimulated hemocytes compared with the control group. We also knocked down the EsTNFR gene with RNAi technology. The results showed that the expression level of these genes related to cell proliferation was significantly down-regulated compared with the control group when the TNF does not bind TNFR. We used Edu technology to repeat the above experiments and the results were similar. Compared with the control group, the hemocytes stimulated by rTNF showed more significant proliferation, and the proliferation rate was significantly down-regulated after knocking down the EsTNFR gene. Therefore, we indicate that TNF binding TNFR can affect the proliferation of E. sinensis hemocytes, which might be manifested by affecting the expression of some proliferation-related genes.

Exposure to artificial wind increases energy intake and reproductive performance of female Swiss mice (Mus musculus) in hot temperatures.

High temperatures and heatwaves are rapidly emerging as an important threat to many aspects of physiology and behavior in females during lactation. The body's capacity to dissipate heat is reduced by high ambient temperatures, increasing the risk of hyperthermia. Exposure to wind, a pervasive environmental factor for most terrestrial animals, is known to increase heat loss, but its effects on the reproductive performance of small mammals remains unclear. In the present study, the effects of wind on the energy budgets, resting metabolic rate and milk energy output (MEO) were measured in lactating Swiss mice at 21 and 32.5°C. Females kept at 32.5°C had a significantly lower resting metabolic rate, food intake and MEO, and lighter offspring, than those kept at 21°C. However, exposure to wind increased the asymptotic food intake of females kept at 32.5°C by 22.5% (P<0.01), their MEO by 20.7% (P<0.05) and their litter mass by 17.6% (P<0.05). The body temperature of females kept at 32.5°C was significantly higher during lactation than that of females kept at 21°C, but this difference was reduced by exposure to wind. These findings suggest that exposure to wind considerably improves reproductive performance, increasing the fitness of small mammals while undergoing hot temperatures during heatwaves.

A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets.

We present a CRISPR-based multi-gene knockout screening system and toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. We apply this system for systematically identifying not only pairwise but also three-way synergistic therapeutic target combinations and successfully validate double- and triple-combination regimens for suppression of cancer cell growth and protection against Parkinson's disease-associated toxicity. This system overcomes the practical challenges of experimenting on a large number of high-order genetic and drug combinations and can be applied to uncover the rare synergistic interactions between druggable targets.