UGT89AC1-mediated quercetin glucosylation is induced upon herbivore damage and enhances Camellia sinensis resistance to insect feeding.

Quercetin is a key flavonol in tea plants (Camellia sinensis (L.) O. Kuntze) with various health benefits, and it often occurs in the form of glucosides. The roles of quercetin and its glucosylated forms in plant defense are generally not well-studied, and remain unknown in the defense of tea. Here, we found higher contents of quercetin glucosides and a decline of the aglucone upon Ectropis grisescens (E. grisescens) infestation of tea. Nine UGTs were strongly induced, among which UGT89AC1 exhibited the highest activity toward quercetin in vitro and in vivo. The mass of E. grisescens larvae that fed on plants with repressed UGT89AC1 or varieties with lower levels of UGT89AC1 was significantly lower than that of larvae fed on controls. Artificial diet supplemented with quercetin glucoside also reduced the larval growth rate, whereas artificial diet supplemented with free quercetin had no significant effect on larval growth. UGT89AC1 was located in both the cytoplasm and nucleus, and its expression was modulated by JA, JA-ILE, and MeJA. These findings demonstrate that quercetin glucosylation serves a defensive role in tea against herbivory. Our results also provide novel insights into the ecological relevance of flavonoid glycosides under biotic stress in plants.

Composition of breast milk from mothers of premature and full-term infants and its influence in Z-Scores for infant physical growth.

BACKGROUND: Breast milk contains various crucial nutrients and biologically active substances and is ideal for newborns. This study aimed to analyze the composition of breast milk from mothers of premature and full-term infants and its influences on the growth of infants. METHODS: Infant-mother dyads examined at our Hospital (March 2016 to May 2017) were included. Milk was collected at 0-1 month, 2-3 months, and 5-6 months and analyzed using a MIRIS human milk analyzer. Z-scores of weight-for-length (WLZ), weight-for-age (WAZ), and length-for-age (LAZ) were calculated. RESULTS: This study included full-term (> 37 weeks of gestation, n = 177) and premature (< 37 weeks, n = 94) infant-mother dyads. The premature infants showed higher ΔWAZ, ΔLAZ, and ΔWLZ from infancy to toddlerhood for the physical growth speed, compared with term infants (P < 0.001). All proteins and true protein components of breast milk decreased with infants' age (P < 0.001). For premature and full-term infants, differences in ΔWAZ and ΔLAZ from birth to infancy and the difference in ΔLAZ, WAZ, and LAZ in toddlerhood were positively associated with non-protein nitrogen (NPN) (all P < 0.05), while the Z-score differences in ΔWLZ from birth to infancy were negatively associated with NPN (all P < 0.05). For premature babies, from birth to infancy stage, ΔWAZ was positively correlated with NPN and carbohydrates while negatively correlated with dry matter (all P < 0.05), and ΔLAZ correlated with NPN (ß = 0.428, P = 0.005). CONCLUSION: Breastfeeding helped premature infants compensatory growth when compared to term infants. Whileduring early infancy stage ΔWLZ gain was negatively associated with increased amounts of NPN in breast milk. This might mean although NPN increase the Z-scores of weight-for-age and length-for-age, with no rise in adipose tissue mass.

Recent Progress Regarding Jasmonates in Tea Plants: Biosynthesis, Signaling, and Function in Stress Responses.

Tea plants have to adapt to frequently challenging environments due to their sessile lifestyle and perennial evergreen nature. Jasmonates regulate not only tea plants' responses to biotic stresses, including herbivore attack and pathogen infection, but also tolerance to abiotic stresses, such as extreme weather conditions and osmotic stress. In this review, we summarize recent progress about jasmonaic acid (JA) biosynthesis and signaling pathways, as well as the underlying mechanisms mediated by jasmontes in tea plants in responses to biotic stresses and abiotic stresses. This review provides a reference for future research on the JA signaling pathway in terms of its regulation against various stresses of tea plants. Due to the lack of a genetic transformation system, the JA pathway of tea plants is still in the preliminary stages. It is necessary to perform further efforts to identify new components involved in the JA regulatory pathway through the combination of genetic and biochemical methods.

An integrated supervision framework to safeguard the urban river water quality supported by ICT and models.

Models and information and communication technology (ICT) can assist in the effective supervision of urban receiving water bodies and drainage systems. Single model-based decision tools, e.g., water quality models and the pollution source identification (PSI) method, have been widely reported in this field. However, a systematic pathway for environmental decision support system (EDSS) construction by integrating advanced single techniques has rarely been reported, impeding engineering applications. This paper presents an integrated supervision framework (UrbanWQEWIS) involving monitoring-early warning-source identification-emergency disposal to safeguard the urban water quality, where the data, model, equipment and knowledge are smoothly and logically linked. The generic architecture, all-in-one equipment and three key model components are introduced. A pilot EDSS is developed and deployed in the Maozhou River, China, with the assistance of environmental Internet of Things (IoT) technology. These key model components are successfully validated via in situ monitoring data and dye tracing experiments. In particular, fluorescence fingerprint-based qualitative PSI and Bayesian-based quantitative PSI methods are effectively coupled, which can largely reduce system costs and enhance flexibility. The presented supervision framework delivers a state-of-the-art management tool in the digital water era. The proposed technical pathway of EDSS development provides a valuable reference for other regions.

(+)-Catechin, epicatechin and epigallocatechin gallate are important inducible defensive compounds against Ectropis grisescens in tea plants.

The tea plant, Camellia sinensis (L.) O. Kuntze, is an economically important, perennial woody plant rich in catechins. Although catechins have been reported to play an important role in plant defences against microbes, their roles in the defence of tea plants against herbivores remain unknown. In this study, we allowed the larvae of Ectropis grisescens, a leaf-feeding pest, to feed on the plants, and alternatively, we wounded the plants and then treated them with E. grisescens oral secretions (WOS). Both approaches triggered jasmonic acid-, ethylene- and auxin-mediated signalling pathways; as a result, plants accumulated three catechin compounds: (+)-catechin, epicatechin and epigallocatechin. Not only was the mass of E. grisescens larvae fed on plants previously infested with E. grisescens or treated with WOS significantly lower than that of larvae fed on controls, but also artificial diet supplemented with epicatechin, (+)-catechin or epigallocatechin gallate reduced larval growth rates. In addition, the exogenous application of jasmonic acid, ethylene or auxin induced the biosynthesis of the three catechins, which, in turn, enhanced the resistance of tea plants to E. grisescens, leading to the coordination of the three signalling pathways. Our results suggest that the three catechins play an important role in the defences of tea plants against E. grisescens.

Indole primes defence signalling and increases herbivore resistance in tea plants.

Upon herbivore attack, plants emit herbivore-induced plant volatiles (HIPVs). HIPVs can prime defences and resistance of intact plants. However, how HIPVs are decoded and translated into functional defence responses is not well understood, especially in long-lived woody plants. Here, we investigated the impact of the aromatic HIPV indole on defence-related early signalling, phytohormone accumulation, secondary metabolite biosynthesis and herbivore resistance in tea plants. We find that tea plants infested with tea geometrid caterpillars release indole at concentrations >450 ng*hr-1 . Exposure to corresponding doses of synthetic indole primes the expression of early defence genes involved in calcium (Ca2+ ) signalling, MPK signalling and jasmonate biosynthesis. Indole exposure also primes the production of jasmonates and defence-related secondary metabolites. These changes are associated with higher herbivore resistance of indole-exposed tea plants. Chemical inhibition of Ca2+ and jasmonate signalling provides evidence that both are required for indole-mediated defence priming and herbivore resistance. Our systematic assessment of the impact of indole on defence signalling and deployment shows that indole acts by boosting Ca2+ signalling, resulting in enhanced jasmonate-dependent defence and resistance in a woody plant. Our work extends the molecular basis of HIPV-induced defence priming from annual plants to an economically important tree species.

Herbivore-induced volatiles influence moth preference by increasing the ß-Ocimene emission of neighbouring tea plants.

Herbivore-induced plant volatiles prime neighbouring plants to respond more strongly to subsequent attacks. However, the key volatiles that trigger this state and their priming mechanisms remain largely unknown. The tea geometrid Ectropis obliqua is one of the most devastating leaf-feeding pests of tea plants. Here, plant-plant communication experiments demonstrated that volatiles emitted from tea plants infested by E. obliqua larvae triggered neighbouring plants to release volatiles that repel E. obliqua adult, especially mated females. Volatile analyses revealed that the quantity of eight volatiles increased dramatically when plants were exposed to volatiles emitted by infested tea plants, including (Z)-3-hexenol, linalool, α-farnesene, ß-Ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). The results of behavioural bioassays demonstrated that ß-Ocimene strongly repelled mated E. obliqua females. Individual volatile compound exposure experiments revealed that (Z)-3-hexenol, linalool, α-farnesene and DMNT triggered the emission of ß-Ocimene from tea plants. Chemical inhibition experiments demonstrated that the emission of ß-Ocimene induced by (Z)-3-hexenol, linalool, α-farnesene and DMNT were dependent on Ca2+ and JA signalling. These findings help us to understand how E. obliqua moths respond to volatiles emitted from tea plants and provide new insight into volatile-mediated plant-plant interactions. They have potential significance for the development of novel insect and pest control strategies in crops.

MOF influences meiotic expansion of H2AX phosphorylation and spermatogenesis in mice.

Three waves of H2AX phosphorylation (γH2AX) have been observed in male meiotic prophase I: the first is ATM-dependent and occurs at leptonema, while the second and third are ATR-dependent, occuring at zygonema and pachynema, respectively. The third wave of H2AX phosphorylation marks and silences unsynapsed chromosomes. Little is known about H2AX phosphorylation expands to chromatin-wide regions in spermatocytes. Here, we report that histone acetyltransferase (HAT) MOF is involved in all three waves of H2AX phosphorylation expansion. Germ cell-specific deletion of Mof in spermatocytes by Stra8-Cre (Mof cKO) caused global loss of H4K16ac. In leptotene and zygotene spermatocytes of cKO mice, the γH2AX signals were observed only along the chromosomal axes, and chromatin-wide H2AX phosphorylation was lost. In almost 40% of early-mid pachytene spermatocytes from Mof cKO mice, γH2AX and MDC1 were detected along the unsynapsed axes of the sex chromosomes, but failed to expand, which consequently caused meiotic sex chromosome inactivation (MSCI) failure. Furthermore, though RAD51 was proficiently recruited to double-strand break (DSB) sites, defects in DSB repair and crossover formation were observed in Mof cKO spermatocytes, indicating that MOF facilitates meiotic DSB repair after RAD51 recruitment. We propose that MOF regulates male meiosis and is involved in the expansion of all three waves of H2AX phosphorylation from the leptotene to pachytene stages, initiated by ATM and ATR, respectively.

The Laccase Gene Family Mediate Multi-Perspective Trade-Offs during Tea Plant (Camellia sinensis) Development and Defense Processes.

Laccase (LAC) plays important roles in different plant development and defense processes. In this study, we identified laccase genes (CsLACs) in Camellia sinensis cv 'Longjing43' cultivars, which were classified into six subclades. The expression patterns of CsLACs displayed significant spatiotemporal variations across different tissues and developmental stages. Most members in subclades II, IV and subclade I exhibited contrasting expression patterns during leaf development, consistent with a trade-off model for preferential expression in the early and late developmental stages. The extensive transcriptional changes of CsLACs under different phytohormone and herbivore treatment were observed and compared, with the expression of most genes in subclades I, II and III being downregulated but genes in subclades IV, V and VI being upregulated, suggesting a growth and defense trade-off model between these subclades. Taken together, our research reveal that CsLACs mediate multi-perspective trade-offs during tea plant development and defense processes and are involved in herbivore resistance in tea plants. More in-depth research of CsLACs upstream regulation and downstream targets mediating herbivore defense should be conducted in the future.

Biallelic Mutations in CFAP43 and CFAP44 Cause Male Infertility with Multiple Morphological Abnormalities of the Sperm Flagella.

Sperm motility is vital to human reproduction. Malformations of sperm flagella can cause male infertility. Men with multiple morphological abnormalities of the flagella (MMAF) have abnormal spermatozoa with absent, short, coiled, bent, and/or irregular-caliber flagella, which impair sperm motility. The known human MMAF-associated genes, such as DNAH1, only account for fewer than 45% of affected individuals. Pathogenic mechanisms in the genetically unexplained MMAF remain to be elucidated. Here, we conducted genetic analyses by using whole-exome sequencing and genome-wide comparative genomic hybridization microarrays in a multi-center cohort of 30 Han Chinese men affected by MMAF. Among them, 12 subjects could not be genetically explained by any known MMAF-associated genes. Intriguingly, we identified compound-heterozygous mutations in CFAP43 in three subjects and a homozygous frameshift mutation in CFAP44 in one subject. All of these recessive mutations were parentally inherited from heterozygous carriers but were absent in 984 individuals from three Han Chinese control populations. CFAP43 and CFAP44, encoding two cilia- and flagella-associated proteins (CFAPs), are specifically or preferentially expressed in the testis. Using CRISPR/Cas9 technology, we generated two knockout models each deficient in mouse ortholog Cfap43 or Cfap44. Notably, both Cfap43- and Cfap44-deficient male mice presented with MMAF phenotypes, whereas the corresponding female mice were fertile. Our experimental observations on human subjects and animal models strongly suggest that biallelic mutations in either CFAP43 or CFAP44 can cause sperm flagellar abnormalities and impair sperm motility. Further investigations on other CFAP-encoding genes in more genetically unexplained MMAF-affected individuals could uncover novel mechanisms underlying sperm flagellar formation.

The Jasmonic Acid Pathway Positively Regulates the Polyphenol Oxidase-Based Defense against Tea Geometrid Caterpillars in the Tea Plant (Camellia sinensis).

Polyphenol oxidases (PPOs) as inducible defense proteins, contribute to tea (Camellia sinensis) resistance against tea geometrid larvae (Ectropis grisescens), and this resistance has been associated with the jasmonic acid (JA) signaling by testing geometrid performance in our previous work. However, the regulation of PPO-based defense by JA and other hormone signaling underlying these defense responses is poorly understood. Here, we investigated the role of phytohormones in regulating the PPO response to tea geometrids. We profiled levels of defense hormones, PPO activity and CsPPO genes in leaves infested with tea geometrids. Then, hormone levels were manipulated by exogenous application of methyl jasmonate (MeJA), gibberellin acid (GA3), abscisic acid (ABA), JA biosynthesis inhibitors (sodium diethyldithiocarbamate trihydrate, DIECA and salicylhydroxamic acid, SHAM) and GA inhibitor (uniconazole, UNI). Upon geometrid attack, JA levels significantly increased, whereas GA levels notably decreased and ABA level was slightly decreased. And the PPO activity significantly increased in line with the transcript levels of CsPPO2 and CsPPO4 but not CsPPO1. There were an obvious antagonistic cross-talk between JA and GA signals and an association among JA signals, PPO response and herbivore resistance in tea plants. Pretreatment with MeJA increased PPO activity by activating the transcripts of CsPPO2 and CsPPO4, whereas application of JA inhibitor DIECA suppressed PPO activity. GA3 strongly enhanced PPO activity, but ABA did not alter PPO activity. These findings strongly suggest that JA is a central player in PPO-mediated tea resistance against tea geometrids in a manner that prioritizes defense over growth.

JA-Ile-macrolactone 5b Induces Tea Plant (Camellia sinensis) Resistance to Both Herbivore Ectropis obliqua and Pathogen Colletotrichum camelliae.

Jasmonates (JAs), the group of lipid-derived hormones, were found to control the defense responses in a myriad of plants. Meaningfully, the macrolactones of 12-hydroxy jasmonate isoleucine (12OH-JA-Ile) were reported to induce the defensive response of wild tobacco. However, little to nothing has been known about the elicitation effect of JA-Ile-macrolactones on woody plants to harmful organisms, let alone its underlying mechanisms. Here, we first optimized the synthetic routine using mild toxic reagent isobutyl chloroformate instead of ethyl chloroformate for conjugation, and we used acetonitrile (MeCN) instead of ethyl alcohol for the better dissolution of p-toluenesulfonic acid (p-TsOH) to gain JA-Ile-macrolactones. JA-Ile-macrolactone 5b-treated tea plants significantly inhibited the larvae weight gain of Ectropis obliqua larvae and the lesions caused by Colletotrichum camelliae. Furthermore, the expression level of CsOPR3 was significantly upregulated in 5b-treated leaves. Meanwhile, 5b reduced the accumulation of eriodictyol 7-O-glucuronide (EDG) in tea plants, which was confirmed to promote the growth rate of E. obliqua larvae by artificial diet assay. In conclusion, our study proved that the exogenous application of 5b could induce the tea plant resistance both to herbivore E. obliqua and pathogen C. camelliae, and EDG was identified as one of the secondary metabolites that could influence the growth rate of E. obliqua, but it did not directly influence the infection of C. camelliae in vitro. Further research should be carried out to clarify the mechanism through which 5b induces tea plant resistance to C. camelliae.

Genome-Wide Identification of the Tify Gene Family and Their Expression Profiles in Response to Biotic and Abiotic Stresses in Tea Plants (Camellia sinensis).

The TIFY family is a plant-specific gene family that is involved in regulating a variety of plant processes, including developmental and defense responses. The chromosome-level genome of the tea plant (Camellia sinensis) has recently been released, but a comprehensive view of the TIFY family in C. sinensis (the CsTIFY genes) is lacking. The current study performed an extensive genome-wide identification of CsTIFY genes. The phylogenetics, chromosome location, exon/intron structure, and conserved domains of these genes were analyzed to characterize the members of the CsTIFY family. The expression profiles of the CsTIFY genes in four organs were analyzed, and they showed different spatial expression patterns. All CsJAZ genes were observed to be induced by jasmonate acid (JA) and exhibited different responses to abiotic and biotic stresses. Six of seven CsJAZ genes (CsJAZ1, CsJAZ2, CsJAZ3, CsJAZ4, CsJAZ7, and CsJAZ8) were upregulated by mechanical wounding and infestation with the tea geometrid (Ectropis obliqua), while infection with tea anthracnose (Colletotrichum camelliae) primarily upregulated the expression levels of CsJAZ1 and CsJAZ10. In addition, CsJAZs were observed to interact with CsMYC2 and AtMYC2. Therefore, the results of this study may contribute to the functional characterization of the CsTIFY genes, especially the members of the JAZ subfamily, as regulators of the JA-mediated defense response in tea plant.

Fusion Method Combining Ground-Level Observations with Chemical Transport Model Predictions Using an Ensemble Deep Learning Framework: Application in China to Estimate Spatiotemporally-Resolved PM2.5 Exposure Fields in 2014-2017.

Atmospheric chemical transport models (CTMs) have been widely used to simulate spatiotemporally resolved PM2.5 concentrations. However, CTM results are usually prone to bias and errors. In this study, we improved the accuracy of PM2.5 predictions by developing an ensemble deep learning framework to fuse model simulations with ground-level observations. The framework encompasses four machine-learning models, i.e., general linear model, fully connected neural network, random forest, and gradient boosting machine, and combines them by stacking approach. This framework is applied to PM2.5 concentrations simulated by the Community Multiscale Air Quality (CMAQ) model for China from 2014 to 2017, which has complete spatial coverage over the entirety of China at a 12-km resolution, with no sampling biases. The fused PM2.5 concentration fields were evaluated by comparing with an independent network of observations. The R2 values increased from 0.39 to 0.64, and the RMSE values decreased from 33.7 µg/m3 to 24.8 µg/m3. According to the fused data, the percentage of Chinese population residing under the level II National Ambient Air Quality Standards of 35 µg/m3 for PM2.5 has increased from 46.5% in 2014 to 61.7% in 2017. The method is readily adapted to utilize near-real-time observations for operational analyses and forecasting of pollutant concentrations and can be extended to provide source apportionment forecasts as well.

Enhanced transcriptome responses in herbivore-infested tea plants by the green leaf volatile (Z)-3-hexenol.

Green leaf volatiles (GLVs) play a vital role in enhancing herbivore-associated defense responses, but the mechanism by which they precisely regulate such responses is not well understood. (Z)-3-Hexenol (z3HOL), an important component of GLVs, effectively activates the defense of tea plants (Camellia sinensis) against a tea geometrid (TG) Ectropis obliqua Prout. To elucidate the molecular mechanisms of defense activation by z3HOL, RNA-Sequencing was employed to investigate the effect of z3HOL on transcriptome responses to TG in tea plants. A total of 318 upregulated genes were identified, and expression of 10 unigenes was validated by quantitative real-time PCR. Among these 318 upregulated genes, 56 were defense-related, including 6 key enzyme genes in jasmonic acid, and ethylene biosynthesis, 24 signal transduction genes, and 12 insect-responsive transcription factors. Most of the defense-related genes are induced by JA, TG, or wounding treatments, suggesting that JA signaling plays a vital role in z3HOL-induced tea defense against TG.

The effect characteristics of temperature on stroke mortality in Inner Mongolia and globally.

The current study investigated the correlation between stroke mortality and temperature. Monthly and seasonal variations in stroke mortality were plotted and daily stroke-related deaths were calculated. The lag times were calculated using the time series analysis. The correlation between stroke incidence and the diurnal temperature range (DTR) was analyzed using case-crossover analysis. Global stroke mortality was described in five latitudes. In the eastern region of Inner Mongolia, the stroke mortality was 174.18/105, about twice of that of the midwestern regions (87.07/105), and temperature was negatively correlated with stroke mortality. Mortality peaked in the winter and troughed in the summer (χ2 = 13.634, P < 0.001). The days in which stroke-related deaths were greater than ten occurred between late October and early April. The effect of temperature on stroke incidence occurred during a lag time of 1 (P = 0.024) or 2 months (P = 0.039). A DTR over 13 °C was positively correlated (r = 0.95, P = 0.004) with stroke with a lag time of 1 day. The effect of temperature on stroke was shown to be the same for various populations. As the latitude increases, stroke mortality also increases with latitudes > 40°; the highest mortality was 188.05/105 at the highest latitude. Only in relatively cold regions as the temperature decreases does stroke mortality increase for various populations. Differences in the time lag as well as in the DTR lag and DTR critical point vary for both the temperature and region.

Two New Polyphenol Oxidase Genes of Tea Plant (Camellia sinensis) Respond Differentially to the Regurgitant of Tea Geometrid, Ectropis obliqua.

Polyphenol oxidases (PPOs) have been reported to play an important role in protecting plants from attacks by herbivores. Though PPO genes in other plants have been extensively studied, research on PPO genes in the tea plant (Camellia sinensis) is lacking. In particular, which members of the PPO gene family elicit the defense response of the tea plant are as yet unknown. Here, two new PPO genes, CsPPO1 and CsPPO2, both of which had high identity with PPOs from other plants, were obtained from tea leaves. The full length of CsPPO1 contained an open reading frame (ORF) of 1740 bp that encoded a protein of 579 amino acids, while CsPPO2 contained an ORF of 1788 bp that encoded a protein of 595 amino acids. The deduced CsPPO1 and CsPPO2 proteins had calculated molecular masses of 64.6 and 65.9 kDa; the isoelectric points were 6.94 and 6.48, respectively. The expression products of recombinant CsPPO1 and CsPPO2 in Escherichia coli were about 91 and 92 kDa, respectively, but the recombinant proteins existed in the form of an inclusion body. Whereas CsPPO1 is highly expressed in stems, CsPPO2 is highly expressed in roots. Further results showed that the expression of CsPPO1 and CsPPO2 was wound- and Ectropis obliqua-induced, and that regurgitant, unlike treatment with wounding plus deionized water, significantly upregulated the transcriptional expression of CsPPO2 but not of CsPPO1. The difference between regurgitant and wounding indicates that CsPPO2 may play a more meaningful defensive role against E. obliqua than CsPPO1. Meanwhile, we found the active component(s) of the regurgitant elicited by the expression of CsPPO may contain small molecules (under 3-kDa molecular weight). These conclusions advance the understanding of the biological function of two new PPO genes and show that one of these, CsPPO2, may be a promising gene for engineering tea plants that are resistant to E. obliqua.

Low-dose lymphocyte immunotherapy rebalances the peripheral blood Th1/Th2/Treg paradigm in patients with unexplained recurrent miscarriage.

BACKGROUND: The published results regarding lymphocytes immunotherapy for unexplained recurrent miscarriage (uRM) patients are conflicting due to different screening criteria and therapeutic protocols. The objective of the present study is to evaluate the effectiveness of immunotherapy using low-dose lymphocytes in patients with uRM and Th1/Th2/Treg paradigm disorders. METHODS: Sixty-four uRM patients who received low-dose lymphocytes immunotherapy served as the immunotherapy group, while the other 35 women who did not receive the treatment served as the control group. The proportions of peripheral blood Th1 cells, Th2 cells and Treg cells; and the concentration of TGF-ß1 in serum were detected by flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively, before and after the immunotherapy. RESULTS: The proportion of Th1 cells was significantly decreased while the proportions of Th2 cells and Treg cells were significantly increased in immunotherapy group patients after treatment. In addition, the concentration of TGF-ß1 in serum was significantly higher after immunotherapy than before. Forty-three uRM patients achieved pregnancy after receiving immunotherapy and 5 patients underwent miscarriages in the immunotherapy group (11.6%, 5/43), while 8 of the 23 pregnant patients experienced a miscarriage in the control group (34.8%, 8/23; p < 0.05). CONCLUSIONS: Low-dose lymphocyte immunotherapy is beneficial for restoring balance in the Th1/Th2/Treg paradigm and improving pregnancy outcome in uRM patients. TRIAL REGISTRATION: NCT03081325 . ClinicalTrials.gov . Retrospectively registered July 2015.

n-Pentacosane Acts as both Contact and Volatile Pheromone in the tea Weevil, Myllocerinus aurolineatus.

Insect cuticular hydrocarbons (CHCs) play important roles in chemical communication, as well as having ecological and physiological roles. The use of CHCs for mate recognition has been shown in many insect genera, but little is known about their use in the tea weevil Myllocerinus aurolineatus. Here, we provide evidence that CHCs on the surface of sexually mature M. aurolineatus females act as contact sex pheromones, facilitating mate recognition and eliciting copulatory behavior in male weevils. Using gas chromatography-mass spectrometry, we identify n-pentacosane and n-heptacosane as two potential contact pheromone components. Results from arena bioassays showed that n-pentacosane is a component of a contact pheromone of M. aurolineatus. Further results from the Y-tube olfactometer bioassays showed that n-pentacosane also acts as a volatile attractant. Our results greatly improve our understanding of the chemical ecology of M. aurolineatus.