To explore the mechanism of acupoint application in the treatment of primary dysmenorrhea by 16S rDNA sequencing and metabolomics.

Graphene-based warm uterus acupoint paste (GWUAP) is an emerging non-drug alternative therapy for the treatment of primary dysmenorrhea (PD), but the underlying mechanism is still unclear. SD female rats were randomly divided into control group, model group and treatment group to explore the mechanism of GWUAP in the treatment of PD. Combined with 16S rDNA and fecal metabolomics, the diversity of microbiota and metabolites in each group was comprehensively evaluated. In this study, GWUAP reduced the torsion score of PD model rats, improved the pathological morphology of uterine tissue, reduced the pathological damage score of uterine tissue, and reversed the expression levels of inflammatory factors, pain factors and sex hormones. The 16 S rDNA sequencing of fecal samples showed that the abundance of Lactobacillus in the intestinal flora of the model group decreased and the abundance of Romboutsia increased, while the abundance of Lactobacillus in the intestinal flora of the treatment group increased and the abundance of Romboutsia decreased, which improved the imbalance of flora diversity in PD rats. In addition, 32 metabolites related to therapeutic effects were identified by metabolomics of fecal samples. Moreover, there is a close correlation between fecal microbiota and metabolites. Therefore, the mechanism of GWUAP in the treatment of PD remains to be further studied.

Acupoint application therapy alleviates pain by regulating immune function through inhibiting TLR4/MyD88/NF-κB p65 signaling in a primary dysmenorrhea rat model. / 穴位贴敷通过TLR4/MyD88/NF-κBé€šè·¯è°ƒèŠ‚åŽŸå‘æ€§ç—›ç»å¤§é¼ çš„å ç–«åŠŸèƒ½.

OBJECTIVES: To investigate the effects of graphene-based warm uterus acupoint paste on uterine Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-kappa B p65 (NF-κB p65) signaling pathway and Th1/Th2 immune balance in primary dysmenorrhea ( PD ) model rats, so as to reveal its immunological mechanisms of relieving dysmenorrhea. METHODS: Thirty SD female rats were randomly divided into 3 groups：normal group, model group and acupoint paste group, with 10 rats in each group. PD rat model was established by subcutaneous injection of estradiol benzoate for 10 consecutive days. At the same time of modeling, graphene-based warm uterus acupoint paste was applied to the acupoints of "Guanyuan" (CV4), bilateral "Zigong" (EX-CA1) and "Sanyinjiao" (SP6) of rats in the acupoint paste group. The application was continuously applied once daily for 10 d, 5 h each time. On the 11th day, oxytocin was injected intraperitoneally to observe the writhing latency, writhing times within 30 min and writhing score of rats in each group. The spleen and thymus indexes were calculated. The pathological changes of spleen and thymus tissue were observed after HE staining. The contents of serum immunoglobulin (Ig) A, IgG, tumor necrosis factor-α (TNF-α), interleukin (IL)-2, interferon-γ (IFN-γ), IL-4 and IL-10 were detected by ELISA . The protein and mRNA expression levels of TLR4, MyD88 and NF-κB p65 in rat uterine tissue were detected by Western blot and real-time quantitative PCR, respectively. RESULTS: Compared with the normal group, the writhing times and writhing scores within 30 min of rats in the model group were significantly increased(P<0.001), and the rats showed writhing reaction (P<0.01). The spleen index and thymus index were significantly decreased(P<0.01, P<0.05). The spleen and thymus had obvious pathological changes. The contents of IgA, IgG, TNF-α, IL-2 and IFN-γ in serum were significantly increased, while the contents of serum IL-4 and IL-10 were significantly decreased(P<0.001, P<0.01). The expression levels of TLR4, MyD88, NF-κB p65 protein and corresponding mRNA in uterine tissue were significantly increased(P<0.001). Following intervention, compared with the model group, the writhing latency time of rats in the acupoint paste group was prolonged, and the writhing times and writhing scores within 30 min were significantly decreased (P<0.001). The spleen index and thymus index were significantly increased(P<0.01, P<0.05). The pathological changes of spleen and thymus were improved. The contents of serum IgA, IgG, TNF-α, IL-2 and IFN-γ were significantly decreased, while the contents of IL-4 and IL-10 were significantly increased(P<0.001, P<0.05, P<0.01). The expression of TLR4, MyD88, NF-κB p65 protein and the corresponding mRNA levels in uterine tissue were decreased(P<0.001, P<0.01). CONCLUSIONS: Graphene-based warm uterus acupoint paste can regulate the immune balance of Th1/ Th2 by regulating TLR4/ MyD88/ NF-κB p65 signaling pathway, repair the pathological damage of immune tissue, improve immune function, and effectively relieve the pain symptoms of PD rats.

Copper-Catalyzed Aryne Insertion into the Carbon-Iodine Bond of Heteroaryl Iodides.

Aryne insertions into the carbon-iodine bond of heteroaryl iodides has been achieved for the first time. This novel reaction provides an efficient pathway for the synthesis of valuable building blocks 2-iodoheterobiaryls from heteroaryl iodides and o-silylaryl triflates in excellent regioselectivity. The copper(I) catalyst, which bears a N-heterocyclic carbene (NHC) ligand, is essential to accomplish the reaction. Control reactions and DFT calculations indicate that the coordination of copper, as a Lewis acid, with nitrogen atoms of heteroaryl iodides mediates the insertion of arynes into heteroaryl carbon-iodine bonds.

Genetic diversity and temporal changes of an endemic cyprinid fish species, Ancherythroculter nigrocauda, from the upper reaches of Yangtze River.

Small populations with low genetic diversity are prone to extinction. Knowledge on the genetic diversity and structure of small populations and their genetic response to anthropogenic effects are of critical importance for conservation management. In this study, samples of Ancherythroculter nigrocauda, an endemic cyprinid fish from the upper reaches of Yangtze River, were collected from five sites to analyze their genetic diversity and population structure using mitochondrial cytochrome b gene and 14 microsatellite loci. Haplotype diversity, nucleotide diversity, and expected heterozygosity indicated that the A. nigrocauda populations had low genetic diversity, and decreased heavily from 2001 to 2016. Significant genetic differentiation was found among different populations in the cyt b gene and SSR markers based on the genetic differentiation index (F ST), whereas no differentiation was found in 2001. Haplotype genealogy showed that eight out of 15 haplotypes were private to one population. The SSR STRUCTURE analysis showed that there were four genetic clusters in the A. nigrocauda samples, with each population forming a single cluster, except for the Chishui River (CSR) and Mudong River (MDR) populations, which formed a common cluster. Therefore, loss of genetic diversity and increased genetic differentiation were found in the A. nigrocauda populations, which could be attributed to dam construction, overfishing, and water pollution in the upper Yangtze River. It is therefore recommended that the government should ban fishing, control water pollution, increase river connectivity, and establish artificial breeding and stocking.

Synthesis, Anti-inflammatory Activities and Mechanisms of 3,5- dihydroxycinnamic Acid Derivatives.

BACKGROUND: 3,4-dihydroxycinnamic acid and its derivatives exhibit numerous biologic activities. Such activities have not previously been reported for 3,5-dihydroxycinnamic acid derivatives. In this study, ten derivatives of 3,5- dihydroxycinnamic acid were synthesized and their anti-inflammatory activities were tested in 12-O-tetradecanoylphorbol 13-acetate-induced mouse ear edema. Molecular biological studies have shed lights on their anti-inflammatory mechanism. METHODS: Anti-inflammatory activities of ten new synthesized derivatives of 3,5-dihydroxycinnamic acid were tested in 12-O-tetradecanoylphorbol 13-acetate-induced mouse ear edema, and their anti-inflammatory mechanism was studied by ELISA, real-time RT-PCR, MPO assay and AA-induced mouse ear edema. RESULTS: Compound 7 showed a pronounced anti-inflammatory effect and the inhibition rate was 65.6% at a dose of 1.6mg/ear. This compound acted by reducing mRNA and protein synthesis of tumor necrosis factor-α, interleukins 1ß and 6, and also by decreasing the levels of activated neutrophil infiltrates. Furthermore, compound 7 significantly suppressed arachidonic acid-induced edema as well. Cell-based assays showed that compound 7 inhibited the production of cyclooxygenase- 2-catalyzed prostaglandin E2 from lipopolysaccharide-treated RAW 264.7 cells, and also inhibited 5-lipoxygenase production from A23187-treated RBL-1 cells, and consequently reduced leukotriene B4 production. CONCLUSION: This investigation revealed that some of the derivatives of 3,5-dihydroxycinnamic acid exhibit a more pronounced anti-inflammatory effect than 3,4-dihydroxycinnamic acid. Therefore, 3,5-dihydroxycinnamic acid derivatives, especially compound 7, represent potential value for antiinflammatory drug development.

Seasonal variation in drifting eggs and larvae in the upper Yangtze, China.

From 5 March to 25 July 2008, ichthyoplankton drifting into the Three Gorges Reservoir from the upper reaches of the Yangtze River were sampled daily to investigate the species composition, abundance, and seasonal variation in early-stage fishes in this area. Twenty-eight species belonging to five orders and 17 families or subfamilies were identified by analyzing fish eggs and larvae, and a total of 14.16 billion individuals were estimated drifting through the sampling section during the investigation. Among the ichthyoplankton sampled, species in Cultrinae, Cobitidae, Gobioninae and Gobiidae, along with the common carp (Cyprinus carpio Linnaeus), comprised 89.6% of the total amount. Six peaks of drift density were identified during the sampling period, and a significant correlation was found between drift density with water discharge. The dominant species were different in each drift peak, indicating different spawning times for the major species. The total amount of the four major Chinese carps that drifted through the sampling section was estimated as 0.88 billion, indicating an increase in the population sizes of these species in the upper reaches of the Yangtze River after construction of the Three Gorges Dam. Actually, these reaches have become the largest spawning area for the four major Chinese carps in the Yangtze River. The large total amount of eggs and larvae drifting through this section demonstrated that the upper reaches of the Yangtze River provided important spawning sites for many fish species, and that conservation of this area should be of great concern.

[Species composition, diversity and density of small fishes in two different habitats in Niushan Lake].

This paper studied the spatial distribution of small fishes in a shallow macrophytic lake, Niushan Lake in spring 2003, and its relations with habitat heterogeneity. Based on the macrophyte cover condition, distance from lake shore and water depth, two representative habitat types in the lake were selected. Habitat A was near the shore with dense submersed macrophyte, while habitat B was far from the shore with sparse submersed macrophyte. Small fishes were sampled quantitatively by block net (180 m2), and their densities within the net area were estimated by multiple mark-recapture or Zippin's removal method. The results showed that there were some differences in species composition, biodiversity measurement, and estimated density of small fishes between the two habitats: 1) the catches in habitat A consisted of 14 small fish species from 5 families, among which, benthopelagic species Rhodeus ocellatus, Paracheilognathus imberbis and Pseudorasbora parva were considered as dominant species, while those in habitat B consisted of 9 small fish species from 3 families, among which, bottom species Rhinogobius giurinus and Micropercops swinhonis were dominant; 2) the Bray-Curtis index between the two small fish communities was 0.222, reflecting their low structure similarity, and no significant difference was observed between their rank/ abundance distributions, both of which belonged to log series distribution; 3) the total density of 9 major species in habitat A was 8.71 ind x m(-2), while that of 5 major species in habitat B was only 3.54 ind x m(-2). The fact that the spatial distribution of the small fishes differed with habitats might be related to their habitat need for escaping predators, feeding, and breeding, and thus, aquatic macrophyte habitat should be of significance in the rational exploitation of small fish resources as well as the conservation of fish resource diversity.

[Process, mechanism, and research method of fish invasion].

Fish invasion has become a serious environmental and economic damage, and obtained a priority concern by the conservation ichthyologists and land managers. A better understanding of the invasive mechanisms is of significance in developing ecological theories and in controlling exotic pests. In this paper, the progress in studying the mechanisms of fish invasion was reviewed, involving the biological characteristics of exotic fish, their adaptive evolution, interactions among exotic and indigenous fish, and relationships between species diversity and community resistance to invasion, etc. The study of invasive process and the progress of research methods were discussed.