Integrated Analysis of Seed microRNA and mRNA Transcriptome Reveals Important Functional Genes and microRNA-Targets in the Process of Walnut (Juglans regia) Seed Oil Accumulation.

Walnut (Juglans regia) is known as a promising woody oil crop with abundant polyunsaturated fatty acids in its kernel. However, the regulation mechanism of walnut oil accumulation and fatty acid metabolism is still poorly understood, which restricted the breeding and genetic improvement of high-quality oil-bearing walnuts. To reveal the molecular mechanism of walnut oil accumulation, considering the potential regulation of microRNA (miRNA) in seed development, in this study, the oil content of walnut kernel on the 80th, 100th and 120th day after flowering (DAF) was tested and the corresponding proportions are 11.51%, 40.40% and 53.20%. Between DAF of 80th~120th, the content of stearic acid and oleic acid tended to increase, but the proportion of other fatty acids tended to decrease. Meanwhile, comparative transcriptome and sRNA-seq analysis on three stages (80th, 100th and 120th DAF), found 204 conserved miRNAs and 554 novel miRNAs in walnut kernels, among which 104 key genes related to walnut oil accumulation were screened. The phospholipid:diacylglycerol acyltransferase metabolic pathway may contribute more to oil accumulation in walnut. 16 miRNA-mRNA regulatory modules related to walnut oil accumulation and fatty acid synthesis were constructed. 8 known miRNAs and 9 novel miRNAs regulate 28 genes involved in fatty acid (FA) metabolism and lipid synthesis. Among them, jre-miRn105, jre-miRn434, jre-miR477d and jre-miR156a.2 are key miRNAs that regulate walnut FA synthesis. Jre-miRn411 and jre-miR399a.1 are closely related to oil accumulation. These data provide new insights and lay the foundation for subsequent studies on walnut FA synthesis and oil accumulation.

Vitamin A supplementation boosts control of antibiotic-resistant Salmonella infection in malnourished mice.

Disseminated disease from non-typhoidal Salmonella enterica strains results in >20% mortality globally. Barriers to effective treatment include emerging multidrug resistance, antibiotic treatment failure, and risk factors such as malnutrition and related micronutrient deficiencies. Individuals in sub-Saharan Africa are disproportionately affected by non-typhoidal S. enterica bloodstream infections. To inform a clinical trial in people, we investigated vitamin A as a treatment in the context of antibiotic treatment failure in a mouse model of vitamin A deficiency. Vitamin A-deficient (VAD) mice exhibited higher systemic bacterial levels with a multidrug-resistant clinical isolate in comparison to mice on a control diet. Sex-specific differences in vitamin A deficiency and disseminated infection with S. enterica serotype Typhimurium (S. Typhimurium) were observed. VAD male mice had decreased weight gain compared to control male mice. Further, infected VAD male mice had significant weight loss and decreased survival during the course of infection. These differences were not apparent in female mice. In a model of disseminated S. Typhimurium infection and antibiotic treatment failure, we assessed the potential of two consecutive doses of vitamin A in alleviating infection in male and female mice on a VAD or control diet. We found that subtherapeutic antibiotic treatment synergized with vitamin A treatment in infected VAD male mice, significantly decreasing systemic bacterial levels, mitigating weight loss and improving survival. These results suggest that assessing vitamin A as a therapy during bacteremia in malnourished patients may lead to improved health outcomes in a subset of patients, especially in the context of antibiotic treatment failure.

Green synthesis of garlic oil nanoemulsion using ultrasonication technique and its mechanism of antifungal action against Penicillium italicum.

Penicillium italicum (P. italicum) can cause significant economic loss of fruits and vegetables. Although garlic oil (GO) is an effective antimicrobial agent, the unstability and hydrophobicity limit its use as an environmentally friendly alternative to the conventional antibiotics against P. italicum. In this study, we focused on the fabrication and characterization of a functional GO nanoemulsion (NE) using ultrasonic technique and revealed the antifungal mechanism of the GO NE on P. italicum based on morphological, structural and molecular analyses. The optimal hydrophilic lipophilic balance (HLB) value determined for GO was 14 through the combination of Tween 80 and Span 80. Then the Box-Benhnken Design (BBD) was applied to produce the GO NE and the effects of different fabrication parameters on the particle size were evaluated. The optimal GO NE was selected with the GO concentration of 5.5%, the Smix concentration of 10%, the ultrasonic time of 5 min and the power of 50%. This GO NE had the smallest particle size of 52.27 nm, the best antifungal effect and the most stability. Furthermore, the antifungal mechanism of the GO NE on P. italicum was evaluated by extracellular conductivity, micro-Raman spectra, fluorescence imaging and scanning electron microscopy (SEM) imaging. The results presented that the GO NE retained the antifungal active ingredients. The fungal cell structure and morphology were malformed after treated with the GO NE and the lipids, nucleic acids and protein of P. italicum were destructed. Finally, the optimal GO NE was applied in vivo and P. italicum in citrus was successfully inhibited. It indicated that the optimal GO NE had the better antifungal activity against P. italicum than the pure GO. Besides, the minimum inhibitory concentration (MIC) of GO after preparing into the NE was changed from 3.7% to 0.01265% with about 300 times improvement of bioavailability. Therefore, the synthetic GO NE which promoted the bioavailability of GO was recommended as a promising alternative to inhibit P. italicum in vegetables and fruits.

[Effect of auricular-point pressure combined with personalized music therapy on perioperative stress response in primipara].

OBJECTIVE: To explore the effect of auricula-point pressure combined with personalized music therapy on perioperative stress response in primipara. METHODS: A total of 1000 primiparas who were elective for cesarean section were randomly divided into an observation group and a control group, 500 cases in each one. 1 h before surgery and 3 h, 6 h, 12 h and 24 h after surgery, individualized music relaxation intervention was given for 1 h in the two groups. On the basis of the treatment, auricular-point pressure was added at Pizhixia (AT4), Shenmen (TF4), Xin (CO15), Zigong (uterus), Luanchao (ovary), Neishengzhiqi (TF2) in the observation group. Each point was pressed for 1 min each time, repeated once every 15 min, the force is from light to heavy, and it is better to have a feeling of distension, sourness and radiation in the auricle. The scores of anxiety self-rating scale, visual analog scale (VAS), heart rate, systolic blood pressure, thyroid stimulating hormone, cortisol and blood glucose levels were observed in the two groups. RESULTS: A total of 973 primiparas completed the clinical study in the end, including 488 patients in the observation group and 485 patients in the control group. Compared to enrollment, the scores of anxiety self-rating scale before entering the operating room in the two groups were significantly improved (P<0.05), and the observation group was superior to the control group (P<0.05). Entering the operating room and tracheal extubation, the heart rate and systolic blood pressure of the primiparas in the observation group were more stable than those in the control group, and the differences were statistically significant (P<0.05). Entering the operating room, 1 h into the surgery and 3 h after surgery, the thyroid stimulating hormone, cortisol and blood glucose levels of the primiparas in the observation group were significantly better than those in the control group (P<0.05). The VAS scores in the observation group at 6 h, 12 h and 24 h after surgery were lower than those in the control group (P<0.05). CONCLUSION: Auricular-point pressure combined with personalized music therapy can reduce the perioperative stress response in primipara and promote the improvement of psychological, physiological and emotional.

[Effect of auricular acupuncture on postpartum rehabilitation of primipara with cesarean].

OBJECTIVE: To explore the postoperative effect of auricular acupuncture on primipara with cesarean in order to promote postpartum rehabilitation. METHODS: A total of 120 primiparas with cesarean were randomly divided into an observation group and a control group, 60 cases in each group. The routine treatment and care after surgery were given in the two groups. On the basis treatment, auricular acupuncture was applied at penqiang (TF5), shenmen (TF4), jiaogan (AH6a), pizhixia (AT4), neishengzhiqi(TF2), pi(CO13), wei(CO4) in the observation group, even-needling technique at all points. The needles were retained for 30 min, 3 times a day, the acupuncture was alternately every other day for 5 days continuously. The postpartum pain, anus exhaust time, incidence of postpartum hemorrhage, urinary retention and constipation, and postpartum average hospitalization day were recorded and compared between the two groups. RESULTS: Twenty-seven cases of postpartum uterine contraction pain (45.0%), 25 cases of wound pain (41.7%), 19 cases of breast distending pain (31.7%) and 11 cases of muscle soreness (18.3%) in the observation group, which was significantly lower than 38 cases (63.3%), 36 cases (60.0%), 30 cases (50.0%) and 21 cases (35.0%) respectively in the control group (P<0.05). The anus exhaust time in the observation group was earlier than that in the control group (P<0.05), and the average hospitalization time was shorter than that in the control group (P<0.05). The incidence of postpartum hemorrhage, urinary retention and constipation in the observation group was lower than that in the control group (P<0.05). CONCLUSION: Auricular acupuncture can effectively relieve postpartum pain in primipara with cesarean, promote postpartum exhaust and shorten hospitalization days, and reduce the incidence of postpartum complications such as postpartum hemorrhage, urinary retention and constipation, it is conducive to postpartum rehabilitation.

[Effects of the combined therapy of the auricular-point pressure at the free position and the unprotected perineal delivery technique during the second stage of labor in the primiparas].

OBJECTIVE: To observe the effects of the combined therapy of the auricular-point pressure at the free position and the unprotected perineal delivery technique during the 2nd stage of labor in the primiparas so as to improve the clinical delivery quality. METHODS: A total of 146 primiparas who accepted the natural delivery willingly were collected. According to the visit sequence, they were divided into an observation group (72 cases) and a control group (74 cases). The free position and the unprotected perineal delivery were adopted in combination during the labor in both of the two groups. Additionally, in the observation group, the auricular-point pressure with semen vaccariae was intervened when entering the active phase. The points were pengqiang (TF5), zigong (uterus), neishengzhiqi (TF2), pizhixia (AT4), shenmen (TF4), jiaogan (AH6a), pi (CO13) and wei (CO4). Each point was pressed for 1 to 2 min each time, repeated once every 10 to 20 min until the end of the 2nd stage of labor. The episiotomy rate, perineal laceration degree, perineal pain degree, the duration of the 2nd labor stage, postpartum hemorrhage, postpartum urine retention and neonatal asphyxia rate were recorded and compared in the primiparas between the two groups. RESULTS: The perineal laceration of the degree Ⅲ and Ⅳ did not occur in the two groups. Compared with the control group, the effect on the perineal laceration was better in the observation group (P<0.05). The episiotomy rate was 5.6% (4/72) in the observation group, lower than that in the control group (P<0.05). The total perineal laceration rate was 68.1% (48/72) in the observation group, similar to the control group (P>0.05). In the observation group, the neonatal asphyxia rate was 1.4% (1/72), the postpartum hemorrhage 6.9% (5/72) and the incidence of urine retention 5.6% (4/72), all lower than 10.8% (8/74), 18.9% (14/74) and 17.6% (13/74) in the control group respectively (all P<0.05). In the observation group, on the 1st, 2nd and 3rd days after labor, the perineal pain degree and the duration of the 2nd stage of labor were superior to the control group (all P<0.001). CONCLUSION: The combined therapy of the auricular-point pressure intervention at the free position and the unprotected perineal delivery technique effectively reduces the episiotomy rate and perineal laceration degree, relieves the perineal pain, reduces the neonatal asphyxia rate and improves the delivery quality during the 2nd stage of labor in the primiparas.

Expression analysis of nine small heat shock protein genes from Tamarix hispida in response to different abiotic stresses and abscisic acid treatment.

Heat shock proteins (HSPs) play important roles in protecting plants against environmental stresses. Furthermore, small heat shock proteins (sHSPs) are the most ubiquitous HSP subgroup with molecular weights ranging from 15 to 42 kDa. In this study, nine sHSP genes (designated as ThsHSP1-9) were cloned from Tamarix hispida. Their expression patterns in response to cold, heat shock, NaCl, PEG and abscisic acid (ABA) treatments were investigated in the roots and leaves of T. hispida by real-time RT-PCR analysis. The results showed that most of the nine ThsHSP genes were expressed at higher levels in roots than in leaves under normal growth condition. All of ThsHSP genes were highly induced under conditions of cold (4 °C) and different heat shocks (36, 40, 44, 48 and 52 °C). Under NaCl stress, all nine ThsHSPs genes were up-regulated at least one stress time-point in both roots and leaves. Under PEG and ABA treatments, the nine ThsHSPs showed various expression patterns, indicating a complex regulation pathway among these genes. This study represents an important basis for the elucidation of ThsHSP gene function and provides essential information that can be used for stress tolerance genetic engineering in future studies.