Growth and Physiological Response of Viola tricolor L. to NaCl and NaHCO3 Stress.

Soil salinization is an important environmental problem worldwide and has a significant impact on the growth of plants. In recent years, the mechanisms of plant salt tolerance have received extensive attention from researchers. In this paper, an experiment was implemented to assess the potential effect of different NaCl and NaHCO3 (sodium bicarbonate-an alkaline salt) concentrations (25 mmol·L-1, 50 mmol·L-1, 100 mmol·L-1, 150 mmol·L-1 and 200 mmol·L-1) on the growth, antioxidant enzymes, osmoprotectants, photosynthetic pigments and MDA of Viola tricolor L. to reveal the physiological response and explore the maximum concentrations of NaCl and NaHCO3 stress that V. tricolor can tolerate. The results showed that NaCl and NaHCO3 treatments had significant effects on osmoprotectants, antioxidant enzymes, photosynthetic pigments, MDA content and the plant height growth of V. tricolor. On day 14 of the NaCl and NaHCO3 stress, the height growth of V. tricolor was significantly greater than CK when the concentration of NaCl and NaHCO3 was less than 100 mmol·L-1. Soluble protein (SP) was significantly greater than CK when the NaCl concentration was less than 150 mmol·L-1 and the NaHCO3 concentration was less than 200 mmol·L-1; soluble sugar (SS) was significantly greater than CK under all NaCl and NaHCO3 treatments; proline (Pro) was significantly greater than CK when the NaCl concentration was 150 mmol·L-1 and the NaHCO3 concentration were 150 and 200 mmol·L-1, respectively. Peroxidase (POD) was significantly greater than CK when the NaCl concentration was less than 200 mmol·L-1 and the NaHCO3 concentration was less than 150 mmol·L-1; superoxide dismutase (SOD) was significantly greater than CK when the NaCl concentration was 50 mmol·L-1 and the NaHCO3 concentrations were 50, 100 and 150 mmol·L-1, respectively; catalase (CAT) was significantly greater than CK when the NaCl and NaHCO3 concentrations were 25, 50 and 100 mmol·L-1, respectively. Chlorophyll (Chl) was significantly lower than CK when the NaCl and NaHCO3 concentrations were greater than 100 mmol·L-1. Malondialdehyde (MDA) gradually increased with the increase in the NaCl and NaHCO3 concentrations. Membership function analysis showed that the concentrations of NaCl and NaHCO3 that V. tricolor was able to tolerate were 150 mmol·L-1 and 200 mmol·L-1, respectively. Beyond these thresholds, osmoprotectants and antioxidant enzymes were seriously affected, Chl degradation intensified, the photosynthetic system was seriously damaged, and the growth of V. tricolor was severely affected. According to a comprehensive ranking of results, the degree of NaCl stress on V. tricolor was lower than that from NaHCO3 when the treatment concentration was lower than 50 mmol·L-1, but higher than that from NaHCO3 when it exceeded 50 mmol·L-1.

Effect of Plant Growth Regulators on Osmotic Regulatory Substances and Antioxidant Enzyme Activity of Nitraria tangutorum.

Plant growth regulators (PGRs) are natural hormones and synthetic hormone analogues. At low concentrations, PGRs have the ability to influence cell division, cell expansion, and cell structure and function, in addition to mediating environmental stress. In this study, experiments were conducted to determine how exogenous PGRs indole acetic acid (IAA), abscisic acid (ABA), and gibberellic acid (GA) influenced osmotic regulatory substances and activity of antioxidant enzymes in Nitraria tangutorum. Using a completely randomized design, IAA, ABA, and GA3 were applied as foliar spray at concentrations of 50 mg/L, 100 mg/L, 150 mg/L, and 200 mg/L to N. tangutorum shrubs. Some selected shrubs did not receive any treatment and served as the control (Ck). The results showed that the foliar spray of IAA, ABA, and GA3 significantly increased the content of osmotic regulatory substances (soluble sugar, soluble protein, and proline) and antioxidant enzymes (SOD and POD) at most concentrations. In addition, the malondialdehyde (MDA) content significantly reduced after treatment, but after regrowth of coppiced shrubs, lipid peroxidation increased and was still lower than Ck. Our study provides evidence that 100 mg/L 150 mg/L, and 200 mg/L concentrations of IAA, ABA, and GA3 treatments are effective for enhancing osmotic regulatory substances and the activity of antioxidant enzymes in N. tangutorum, which offers an effective strategy not only for increasing tolerance to abiotic and biotic stresses, but also improving the adaptability of N. tangutorum shrubs to the environment.

Gene Regulation during Carapacial Ridge Development of Mauremys reevesii: The Development of Carapacial Ridge, Ribs and Scutes.

The unique topological structure of a turtle shell, including the special ribs-scapula relationship, is an evolutionarily novelty of amniotes. The carapacial ridge is a key embryonic tissue for inducing turtle carapace morphologenesis. However, the gene expression profiles and molecular regulatory mechanisms that occur during carapacial ridge development, including the regulation mechanism of rib axis arrest, the development mechanism of the carapacial ridge, and the differentiation between soft-shell turtles and hard-shell turtles, are not fully understood. In this study, we obtained genome-wide gene expression profiles during the carapacial ridge development of Mauremys reevesii using RNA-sequencing by using carapacial ridge tissues from stage 14, 15 and 16 turtle embryos. In addition, a differentially expressed genes (DEGs) analysis and a gene set enrichment analysis (GSEA) of three comparison groups were performed. Furthermore, a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was used to analyze the pathway enrichment of the differentially expressed genes of the three comparative groups. The result displayed that the Wnt signaling pathway was substantially enriched in the CrTK14 vs. the CrTK15 comparison group, while the Hedgehog signaling pathway was significantly enriched in the CrTK15 vs. the CrTK16 group. Moreover, the regulatory network of the Wnt signaling pathway showed that Wnt signaling pathways might interact with Fgfs, Bmps, and Shh to form a regulatory network to regulate the carapacial ridge development. Next, WGCNA was used to cluster and analyze the expression genes during the carapacial ridge development of M. reevesii and P. sinensis. Further, a KEGG functional enrichment analysis of the carapacial ridge correlation gene modules was performed. Interesting, these results indicated that the Wnt signaling pathway and the MAPK signaling pathway were significantly enriched in the gene modules that were highly correlated with the stage 14 and stage 15 carapacial ridge samples of the two species. The Hedgehog signaling pathway was significantly enriched in the modules that were strongly correlated with the stage 16 carapacial ridge samples of M. reevesii, however, the PI3K-Akt signaling and the TGF-ß signaling pathways were significantly enriched in the modules that were strongly correlated with the stage 16 carapacial ridge samples of P. sinensis. Furthermore, we found that those modules that were strongly correlated with the stage 14 carapacial ridge samples of M. reevesii and P. sinensis contained Wnts and Lef1. While the navajo white 3 module which was strongly correlated with the stage 16 carapacial ridge samples of M. reevesii contained Shh and Ptchs. The dark green module strongly correlated with the stage 16 carapacial ridge samples of P. sinensis which contained Col1a1, Col1a2, and Itga8. Consequently, this study systematically revealed the signaling pathways and genes that regulate the carapacial ridge development of M. reevesii and P. sinensis, which provides new insights for revealing the molecular mechanism that is underlying the turtle's body structure.

RNAi silencing of the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene inhibits vitellogenesis in Chinese mitten crab Eriocheir sinensis.

The sesquiterpenoid methyl farnesoate (MF), a de-epoxide form of insect juvenile hormone III (JH III), plays an essential role in regulating many crucial physiological processes in crustaceans including vitellogenesis and reproduction. 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is an important rate-limiting enzyme in the mevalonate pathway, which is critical for the synthesis of JH III and MF. In the present study, a full-length cDNA encoding HMGR (EsHMGR) in Eriocheir sinensis was isolated and characterised. Sequence analysis of EsHMGR revealed that it belongs to Class I HMGR family proteins with HMG-CoA-binding and NADPH-binding domains, both important for HMGR activity. In addition to its ubiquitous tissue expression, expression of EsHMGR was highly specific to the ovary, the main site of Vg synthesis. During ovarian development, EsHMGR expression in ovary displayed a stage-specific pattern, and was correlated with expression of vitellogenin (EsVg) in hepatopancreas, which suggests that EsHMGR possibly involved in vitellogenesis. To further investigate the functional role of EsHMGR in vitellogenin biosynthesis in E. sinensis, RNA interference-mediated gene silencing was carried out both in vitro and in vivo. Quantitative PCR results showed that injection of EsHMGR double-stranded RNA (dsRNA) led to a significant decrease in EsVg expression levels in ovary and hepatopancreas both in vitro and in vivo. Taken together, the results suggest that EsHMGR is involved in vitellogenin biosynthesis in female E. sinensis, which may provide a new resource for HMGR enzymes participating in reproduction in crustaceans.

Global Analysis of Transcriptome and Translatome Revealed That Coordinated WNT and FGF Regulate the Carapacial Ridge Development of Chinese Soft-Shell Turtle.

The turtle carapace is composed of severely deformed fused dorsal vertebrae, ribs, and bone plates. In particular, the lateral growth in the superficial layer of turtle ribs in the dorsal trunk causes an encapsulation of the scapula and pelvis. The recent study suggested that the carapacial ridge (CR) is a new model of epithelial-mesenchymal transition which is essential for the arrangement of the ribs. Therefore, it is necessary to explore the regulatory mechanism of carapacial ridge development to analyze the formation of the turtle shell. However, the current understanding of the regulatory network underlying turtle carapacial ridge development is poor due to the lack of both systematic gene screening at different carapacial ridge development stages and gene function verification studies. In this study, we obtained genome-wide gene transcription and gene translation profiles using RNA sequencing and ribosome nascent-chain complex mRNA sequencing from carapacial ridge tissues of Chinese soft-shell turtle at different development stages. A correlation analysis of the transcriptome and translatome revealed that there were 129, 670, and 135 codifferentially expressed genes, including homodirection and opposite-direction differentially expressed genes, among three comparison groups, respectively. The pathway enrichment analysis of codifferentially expressed genes from the Kyoto Encyclopedia of Genes and Genomes showed dynamic changes in signaling pathways involved in carapacial ridge development. Especially, the results revealed that the Wnt signaling pathway and MAPK signaling pathway may play important roles in turtle carapacial ridge development. In addition, Wnt and Fgf were expressed during the carapacial ridge development. Furthermore, we discovered that Wnt5a regulated carapacial ridge development through the Wnt5a/JNK pathway. Therefore, our studies uncover that the morphogenesis of the turtle carapace might function through the co-operation between conserved WNT and FGF signaling pathways. Consequently, our findings revealed the dynamic signaling pathways acting on the carapacial ridge development of Chinese soft-shell turtle and provided new insights into uncover the molecular mechanism underlying turtle shell morphogenesis.

Molecular evidence for farnesoic acid O-methyltransferase (FAMeT) involved in the biosynthesis of vitellogenin in the Chinese mitten crab Eriocheir sinensis.

Sesquiterpenoid methyl farnesoate (MF), a crustacean equivalent of insect juvenile hormone (JH III), has essential functions in regulating physiological processes in crustaceans, including reproduction and vitellogenesis. Farnesoic acid O-methyltransferase (FAMeT) is a key rate-limiting enzyme catalyzing the conversion of farnesoic acid (FA) to JH/MF in insects and crustaceans. In this study, a full-length cDNA of EsFAMeT from Eriocheir sinensis was isolated and characterized. The deduced EsFAMeT amino acid sequence indicated there were two conserved Methyltransf-FA domains characteristic of FAMeT family proteins. With use of sequence alignment analysis procedures, there was an indication that FAMeT proteins are highly conserved among crustaceans and FAMeT is more closely related to crustacean FAMeT than to insect FAMeT. Results from quantitative real-time PCR analysis revealed there was ubiquitous EsFAMeT in all tissues examined, with greater abundances of mRNA transcripts in the ovary. The transcription of EsFAMeT indicated there were stage-specific patterns in the hepatopancreas and ovary during ovarian development, with the greatest abundance during ovarian development Stages II and III, respectively. To investigate functions of EsFAMeT in vitellogenin biosynthesis in E. sinensis, RNA interference-mediated gene knockdown was used in vitro and in vivo. Injection of EsFAMeT dsRNA resulted in a marked decrease in EsVg (encoding vitellogenin) transcripts in the ovary and hepatopancreas both in vitro and in vivo. Results from the present study indicated EsFAMeT is involved in vitellogenin biosynthesis in the ovary and hepatopancreas of E. sinensis, providing a new resource to study modulatory effects of the FAMeT family of enzymes in crustacean reproduction.

Identification, characterization and mRNA transcript abundance profiles of the carboxylesterase (CXE5) gene in Eriocheir sinensis suggest that it may play a role in methyl farnesoate degradation.

The sesquiterpenoid methyl farnesoate (MF) is a de-epoxidized form of insect juvenile hormone (JH) III in crustaceans, and its precise titer plays important roles in regulating many critical physiological processes, including reproduction and ovarian maturation. Understanding the synthetic and degradation pathways of MF is equally important for determining how to maintain MF titers at appropriate levels and thus for potential applications in crab aquaculture. Although the synthetic pathway of MF has been well established, little is known about MF degradation. Previous research proposed that specific carboxylesterases (CXEs) that degrade MF in crustaceans are conserved from those of JH III. In this study, we identified a novel Es-CXE5 gene from Eriocheir sinensis. The Es-CXE5 protein contains some conserved motifs, including catalytic triad and oxyanion hole, which are characteristics of the biologically active CXE family. The phylogenetic analysis showed that Es-CXE5 belongs to the hormone/semiochemical processing group of the CXE family. Moreover, Tissue and stage-specific expression results suggested that Es-CXE5 expression in hepatopancreas was highest and associated with the hemolymph MF titer. Furthermore, Es-CXE5 mRNA transcripts were detected in both in vitro and in vivo experiments and ESA experiment in the hepatopancreas and ovary. The results of this study showed that Es-CXE5 mRNA abundance in the hepatopancreas was notably induced by MF addition but had no effect on the ovary. Taken together, our results suggest that Es-CXE5 may degrade MF in the hepatopancreas and may thus be involved in ovarian development in E. sinensis.

Effects of methyl farnesoate on Krüppel homolog 1 (Kr-h1) during vitellogenesis in the Chinese mitten crab (Eriocheir sinensis).

Methyl farnesoate (MF), a de-epoxidized form of juvenile hormone (JH) Ⅲ in insects, may regulate developmental processes such as reproduction and ovarian maturation in crustaceans. Krüppel homolog 1 (Kr-h1) is a target response gene for the methoprene-tolerant (Met) protein that is a component of the JH signaling pathway in insects. In the present study, Es-Kr-h1 was cloned from E. sinensis and characterized to ascertain whether JH/MF signaling in insects is conserved in crustaceans. The findings with molecular structure analysis indicated Es-Kr-h1 contains seven zinc finger motifs (Zn2-Zn8) commonly conserved in other crustaceans, but the Zn1 motif was not detected to be present. The PCR results indicated that relative abundance of Es-Kr-h1 mRNA transcript in the hepatopancreas was greatest in the Stage Ⅱ, followed by the Stage Ⅳ ovarian developmental categories. The relative abundance of Es-Kr-h1 mRNA transcript in vitro was greater after MF addition to the hepatopancreas, however, not the ovarian tissues. The results from in vivo and eyestalk ablation experiments indicated the relative abundance of Es-Kr-h1 mRNA transcript was greater after MF treatment and bilateral eyestalk removal in the hepatopancreas, however, not ovarian tissues. Notably, there were effects of MF on relative abundance of Es-Kr-h1 mRNA transcript pattern. The Es-Kr-h1 protein, therefore, may be involved in MF-mediated vitellogenesis resulting from the response to Es-Met in E. sinensis, and the JH/MF signaling pathway is potentially conserved in crustaceans.

Potential role of Methoprene-tolerant (Met) in methyl farnesoate-mediated vitellogenesis in the Chinese mitten crab (Eriocheir sinensis).

Methoprene-tolerant (Met) belongs to the basic helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) family of nuclear transcriptional regulators and is a leading candidate receptor for juvenile hormone (JH III) in insects. Methyl farnesoate (MF) is a de-epoxide form of JH III that regulates many developmental processes in crustaceans, including reproduction, molting, and morphogenesis, much like JH III in insects. In this study, the full-length cDNA for Met was cloned from the Chinese mitten crab (Eriocheir sinensis) (EsMet). The amino acid sequence of EsMet contains three conserved domains (bHLH, PAS-A, and PASB) characteristic of the bHLH-PAS family, having six conserved amino acid residues specifically responsible for JH or MF binding. Tissue distribution analysis revealed that EsMet mRNA is highly expressed in the hepatopancreas. In addition, EsMet and EsVg expression in the hepatopancreas were found to be significantly increased in early endogenous vitellogenic oocytes (stage II) during ovarian development, and the hemolymph MF titer was significantly increased in late exogenous vitellogenic oocytes (stage III), indicating that EsMet is involved in vitellogenesis regulation. In vitro, MF addition markedly upregulated EsMet and EsVg expression in hepatopancreatic tissue, but only EsVg was induced in ovarian tissue. In vivo, EsMet and EsVg expression in the hepatopancreas were both significantly and synchronously increased after MF injection, but not in the ovaries. In addition, EsMet and EsVg expression were upregulated in the hepatopancreas after eyestalk ablation, while only EsVg expression was induced in the ovaries. Thus, our results indicate that Met may act as a receptor for MF in MF-mediated vitellogenesis in crustaceans.

Systematic selection and validation of suitable reference genes for quantitative real-time PCR normalization studies of gene expression in Nitraria tangutorum.

Suitable reference genes can be used to calibrate the error in quantitative real-time PCR (qPCR) experiments, making the results more credible. However, there are no reference genes suitable for multiple species and under different experimental conditions. Nitraria tangutorum Bobr. is a typical plant native to desert areas. It is drought-resistant, saline-alkali resistant, extreme temperatures-resistant, and has strong adaptability. To date, the importance of this germplasm has not been sufficiently understood; therefore, it is still unclear which genes can be used as reference genes to calibrate qPCR data of N. tangutorum. In this study we analyzed the expression levels of 10 candidate reference genes (ACT, GAPDH, TUA, TUB, CYP, UBC, His, PP2A, HSP, and EF1-α) in N. tangutorum seedlings under a series of experimental conditions, including in different organs (root, stem, and leaf) and under abiotic stresses (salt, drought, heat, and cold) and hormone stimuli (abscisic acid) by qPCR. Three software programs (geNorm, NormFinder, and BestKeeper) were used to evaluate the expression stability of the ten genes. Comprehensive analysis showed that EF1-α and His had the best expression stability, whereas HSP was the least suitable as a reference gene. The expression profile of NtCER7, a gene related to the regulation of cuticular wax biosynthesis in N. tangutorum, verified the accuracy of the experimental results. Based on this study, we recommend EF1-α and His as suitable reference genes for N. tangutorum. This paper provides the first data on stable reference genes in N. tangutorum, which will be beneficial to studying the gene expression of N. tangutorum and other Nitraria species in the future.

Perturbations of gut microbiota in gestational diabetes mellitus patients induce hyperglycemia in germ-free mice.

Shifts in the maternal gut microbiota have been implicated in the development of gestational diabetes mellitus (GDM). Understanding the interaction between gut microbiota and host glucose metabolism will provide a new target of prediction and treatment. In this nested case-control study, we aimed to investigate the causal effects of gut microbiota from GDM patients on the glucose metabolism of germ-free (GF) mice. Stool and peripheral blood samples, as well as clinical information, were collected from 45 GDM patients and 45 healthy controls (matched by age and prepregnancy body mass index (BMI)) in the first and second trimester. Gut microbiota profiles were explored by next-generation sequencing of the 16S rRNA gene, and inflammatory factors in peripheral blood were analyzed by enzyme-linked immunosorbent assay. Fecal samples from GDM and non-GDM donors were transferred to GF mice. The gut microbiota of women with GDM showed reduced richness, specifically decreased Bacteroides and Akkermansia, as well as increased Faecalibacterium. The relative abundance of Akkermansia was negatively associated with blood glucose levels, and the relative abundance of Faecalibacterium was positively related to inflammatory factor concentrations. The transfer of fecal microbiota from GDM and non-GDM donors to GF mice resulted in different gut microbiota colonization patterns, and hyperglycemia was induced in mice that received GDM donor microbiota. These results suggested that the shifting pattern of gut microbiota in GDM patients contributed to disease pathogenesis.

Identification and functional analysis of SOX transcription factors in the genome of the Chinese soft-shell turtle (Pelodiscus sinensis).

SOX transcription factors play an irreplaceable role in biological developmental processes. Sox genes have been identified in a wide variety of species; however, their identification and functional analysis in the genome of the Chinese soft-shell turtle (Pelodiscus sinensis) have not been performed. In the present study, the Chinese soft-shell turtle genome was found to contain 17 Sox genes, which were categorized into seven groups according to their phylogenetic relationships. Gene structure and protein motif analysis of the Sox genes showed that within the same phylogenetic group, their exon-intron number and motif structure of the Sox family were relatively conserved, but diverged in the comparison between different groups. Sexual dimorphism expression analysis for the Sox genes displayed that Sox8 and Sox9 were upregulated in the testis, while Sox3, Sox7, Sox11, and Sox13 were upregulated in the ovary. A correlation network analysis of SOX transcription factors with their target genes analysis showed that Sox3 correlated negatively with Sox9 and gata4. Sox11 and Sox7 correlated negatively with gata4. Sox8 and Sox9 correlated positively with gata4. Therefore, the genome-wide identification and functional analysis of the Sox gene family will be useful to further reveal the functions of Sox genes in the Chinese soft-shell turtle.

[Hydrological functions of litters under five typical plantations in southern and northern mountains of Lanzhou City, Northwest China]. / å °å·žå¸‚å—åŒ—ä¸¤å±±5ç§å ¸åž‹äººå·¥æž—å‡‹è½ç‰©çš„æ°´æ–‡åŠŸèƒ½.

Combined with field observation and indoor water immersion test, water holding characteristics of litters from five typical plantations (Platycladus orientalis, Robinia pseudoacacia, Populus alba var. pyramidalis, P. orientalis+R. pseudoacacia, P. alba var. pyramidalis+R. pseudoacacia) in southern and northern mountains of Lanzhou City were examined. Results showed that litter mass under the plantations ranged from 13.50 to 47.01 t·hm-2, with an order of P. alba var. pyramidalis+R. pseudoacacia＞P. orientalis+R. pseudoacacia＞P. orientalis＞R. pseudoacacia＞P. alba var. pyramidalis. The percentage of un-decomposed litters was greater than that of semi-decomposed litters in all plantations except for P. orientalis plantations. The maximum water-holding rate of litters ranged from 190.8% to 262.7%, with the greatest value in the P. alba var. pyramidalis+R. pseudoa-cacia and the lowest in P. orientalis plantations. The maximum water-holding capacity of litters was 35.29-123.59 t·hm-2, with an order of P. alba var. pyramidalis+R. pseudoacacia＞P. orientalis+R. pseudoacacia＞R. pseudoacacia＞P. orientalis＞P. alba var. pyramidalis. Litter water absorption rate declined linearly within the first hour, and then decreased slowly. Semi-decomposed litters had a higher water-absorption rate than un-decomposed litters. The maximum water retaining amount and effective retaining amount of the litters were P. alba var. pyramidalis+R. pseudoacacia＞P. orientalis+R. pseudoacacia＞P. orientalis＞R. pseudoacacia＞P. alba var. pyramidalis. P. alba var. pyramidalis+R. pseudoacacia had the highest effective retaining rate. P. alba var. pyramidalis+R. pseudoacacia plantation had highest capacity for soil and water conservation in southern and northern mountains of Lanzhou City.

Expression of the kisspeptin/gonadotropin-releasing hormone (GnRH) system in the brain of female Chinese sucker (Myxocyprinus asiaticus) at the onset of puberty.

The kisspeptin-kisspeptin receptor (kissr)-gonadotropin-releasing hormone (GnRH) system plays a key role in regulating the onset of puberty in mammals. However, the role of this system in fish is still unclear. We examined the relative gene expression patterns for kiss1, kiss2, kissr2, sGnRH, and pjGnRH in all parts of the brains of Chinese sucker (Myxocyprinus asiaticus) females at the prepubertal and pubertal stages by using real-time PCR. We also analyzed the expression of kiss1 and GnRH1 via immunofluorescence. Two variants of kisspeptin; a variant of kissr (kissr2); and two variants of GnRH, pjGnRH (GnRH1), and sGnRH (GnRH3), were expressed in all parts of the brain. The mRNA expression of kiss1 was higher in the telencephalon, mesencephalon, and diencephalon at the pubertal stage than at the prepubertal stage, and the expression of kiss2 was higher in only the telencephalon. The expression of kissr2 was higher in all parts of the brain, except the medulla, at the pubertal stage than at the prepubertal stage. pjGnRH was highly expressed in all parts of the brain at the pubertal stage, whereas sGnRH expression showed no distinct changes, except in the epencephalon. Strong kiss1 and weak GnRH-1 immunoreactivity was observed in the pineal gland, lateral tuberal nucleus (NLT), and ventral part of the NLT in the diencephalon of the Chinese sucker females at the pubertal stage. Our results suggest that the kiss1-kissr2-pjGnRH system was expressed highly at the onset of pubertal female Chinese sucker.

Characterization and phylogenetic analysis of the complete mitochondrial genome of Rhinogobius sp. (Perciformes, Gobiidae).

The genus Rhinogobius was widely distributed in East Asia. In the present study, the complete mitochondrial genome of Rhinogobius sp., possible a new species of freshwater goby from Anhui province of China, was sequenced for the first time. Sequence analysis showed that it is 16,511 bp in length with A + T content of 52.3%, consisting of 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs, and a control region (CR). Phylogenetic analyses placed Rhinogobius sp. in a well-supported monophyletic cluster with other Rhinogobius fish and the phylogenetic position of Rhinogobius sp. was closer to Rhinogobius cliffordpopei.

Screening and characterisation of sex differentiation-related long non-coding RNAs in Chinese soft-shell turtle (Pelodiscus sinensis).

Long non-coding RNAs (lncRNAs) perform distinct functions in various biological processes in mammals, including sex differentiation. However, the roles of lncRNAs in other vertebrates, especially in the Chinese soft-shell turtle (Pelodiscus sinensis), remain to be clarified. In this study, we performed genome-wide analysis of the lncRNA expression profiles in gonad tissues and screened numerous sex-specific lncRNAs in the Chinese soft-shell turtle. Of the 363,310,650 clean reads obtained, 5,994 sequences were typed as lncRNAs, of which 4,463 were novel. A selection of sex-specific lncRNAs (♀ 932, ♂ 449) from female ovaries and male testis were shown to act on target genes in cis and in trans, and most were involved in gonad differentiation based on Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Furthermore, interactions among the differentially expressed lncRNA-mRNAs and protein coding genes were identified by construction of correlation networks. Overall, our systematic analysis of lncRNA expression profiles in gonad tissues revealed numerous sex-specific lncRNAs in P. sinensis. Thereby, these findings provide new insights into the function of lncRNAs in sex differentiation and highlight a group of candidate lncRNAs for future studies.

A randomized clinical trial of exercise during pregnancy to prevent gestational diabetes mellitus and improve pregnancy outcome in overweight and obese pregnant women.

BACKGROUND: Obesity and being overweight are becoming epidemic, and indeed, the proportion of such women of reproductive age has increased in recent times. Being overweight or obese prior to pregnancy is a risk factor for gestational diabetes mellitus, and increases the risk of adverse pregnancy outcome for both mothers and their offspring. Furthermore, the combination of gestational diabetes mellitus with obesity/overweight status may increase the risk of adverse pregnancy outcome attributable to either factor alone. Regular exercise has the potential to reduce the risk of developing gestational diabetes mellitus and can be used during pregnancy; however, its efficacy remain controversial. At present, most exercise training interventions are implemented on Caucasian women and in the second trimester, and there is a paucity of studies focusing on overweight/obese pregnant women. OBJECTIVE: We sought to test the efficacy of regular exercise in early pregnancy to prevent gestational diabetes mellitus in Chinese overweight/obese pregnant women. STUDY DESIGN: This was a prospective randomized clinical trial in which nonsmoking women age >18 years with a singleton pregnancy who met the criteria for overweight/obese status (body mass index 24≤28 kg/m2) and had an uncomplicated pregnancy at <12+6 weeks of gestation were randomly allocated to either exercise or a control group. Patients did not have contraindications to physical activity. Patients allocated to the exercise group were assigned to exercise 3 times per week (at least 30 min/session with a rating of perceived exertion between 12-14) via a cycling program begun within 3 days of randomization until 37 weeks of gestation. Those in the control group continued their usual daily activities. Both groups received standard prenatal care, albeit without special dietary recommendations. The primary outcome was incidence of gestational diabetes mellitus. RESULTS: From December 2014 through July 2016, 300 singleton women at 10 weeks' gestational age and with a mean prepregnancy body mass index of 26.78 ± 2.75 kg/m2 were recruited. They were randomized into an exercise group (n = 150) or a control group (n = 150). In all, 39 (26.0%) and 38 (25.3%) participants were obese in each group, respectively. Women randomized to the exercise group had a significantly lower incidence of gestational diabetes mellitus (22.0% vs 40.6%; P < .001). These women also had significantly less gestational weight gain by 25 gestational weeks (4.08 ± 3.02 vs 5.92 ± 2.58 kg; P < .001) and at the end of pregnancy (8.38 ± 3.65 vs 10.47 ± 3.33 kg; P < .001), and reduced insulin resistance levels (2.92 ± 1.27 vs 3.38 ± 2.00; P = .033) at 25 gestational weeks. Other secondary outcomes, including gestational weight gain between 25-36 gestational weeks (4.55 ± 2.06 vs 4.59 ± 2.31 kg; P = .9), insulin resistance levels at 36 gestational weeks (3.56 ± 1.89 vs 4.07 ± 2.33; P = .1), hypertensive disorders of pregnancy (17.0% vs 19.3%; odds ratio, 0.854; 95% confidence interval, 0.434-2.683; P = .6), cesarean delivery (except for scar uterus) (29.5% vs 32.5%; odds ratio, 0.869; 95% confidence interval, 0.494-1.529; P = .6), mean gestational age at birth (39.02 ± 1.29 vs 38.89 ± 1.37 weeks' gestation; P = .5); preterm birth (2.7% vs 4.4%, odds ratio, 0.600; 95% confidence interval, 0.140-2.573; P = .5), macrosomia (defined as birthweight >4000 g) (6.3% vs 9.6%; odds ratio, 0.624; 95% confidence interval, 0.233-1.673; P = .3), and large-for-gestational-age infants (14.3% vs 22.8%; odds ratio, 0.564; 95% confidence interval, 0.284-1.121; P = .1) were also lower in the exercise group compared to the control group, but without significant difference. However, infants born to women following the exercise intervention had a significantly lower birthweight compared with those born to women allocated to the control group (3345.27 ± 397.07 vs 3457.46 ± 446.00 g; P = .049). CONCLUSION: Cycling exercise initiated early in pregnancy and performed at least 30 minutes, 3 times per week, is associated with a significant reduction in the frequency of gestational diabetes mellitus in overweight/obese pregnant women. And this effect is very relevant to that exercise at the beginning of pregnancy decreases the gestational weight gain before the mid-second trimester. Furthermore, there was no evidence that the exercise prescribed in this study increased the risk of preterm birth or reduced the mean gestational age at birth.

Effect of gonadotropin-releasing hormone vaccination on T lymphocyte changes in male rats.

The aim of this study was to detect the effect of immunization against gonadotropin-releasing hormone (GnRH) on cell-meditated immunity. Three-week-old male Sprague-Dawley rats (n=32) were randomly and equally assigned to two groups: 1) GnRH-tandem-ovalbumin immunized group; and 2) the control group (injected with an equivalent Al(OH)3 adjuvant). Blood samples were collected at two-week intervals to assess the level of GnRH-specific antibodies and testosterone. Moreover, blood and thymus samples were also collected to analyze the T lymphocyte subpopulations one and two months after the last booster immunization. T lymphocyte immunity against GnRH was activated during the first month post-immunization as exhibited by increased numbers of CD3+ (P<0.05) and CD4+ (P<0.05)T lymphocytes following testosterone suppression (P<0.01), which was then restored and maintained at appropriate levels in the second month. In contrast, the differentiation of T lymphocytes in the thymus was reduced during the first month after immunization as exhibited by the significant decreased number of CD3+ (P<0.05) cells, followed by the restoration and heightened numbers at later time points for both the number of CD3+ (P<0.05) and CD4+ (P<0.01)T lymphocytes. These results suggest that immunization against GnRH interferes with the number of lymphocytes during the early time points following immunization. The number of T lymphocytes initially decreased in the peripheral blood following immunization, but was replenished by newly exported cells from the thymus which eventually restored the T lymphocytes to normal levels.

[Early selection of drought-resistant superior families of Reaumuria soongorica based on the analysis of osmoregulation substances and chlorophyll].

By using 77 families of 3 year-old Reaumuria soongorica seedlings that grew well without injection of any pests and diseases as experimental material, the contents of soluble protein (SP), soluble sugar (SS), proline (Pro) and chlorophyll (Chl) were measured, and their drought resistance was evaluated with principal component analysis and subordinate function method. The results showed that there were great differences in leaf osmolytes and chlorophyll among the 77 families. The soluble protein content varied from 2.14 to 8.60 mg · g⁻¹ FM, the soluble sugar content was from 6.82 to 21.86 mg · g⁻¹ FM, the proline content was from 118.73 to 1494.30 µg · g⁻¹ FM, the chlorophyll a content was from 321.88 to 897.37 µg · g⁻¹ FM, the chlorophyll b content was from 53.65 to 249.04 µg · g⁻¹ FM, chlorophyll (a+b) was from 387.39 to 1146.40 µg · g⁻¹ FM, and the chlorophyll a/b was from 3.46 to 6.42. All drought-resistant indices had significant difference among R. soongorica families, among which the proline content varied most, followed by the soluble sugar content. Evaluated by using the synthesized multi-index, it was found that 12 families showed good drought resistance, with Zhazigou 1-2 and Zhazigou 1-1 performing the best.