Marked variations in gut microbial diversity, functions, and disease risk between wild and captive alpine musk deer.

Maintaining a healthy status is crucial for the successful captive breeding of endangered alpine musk deer (Moschus chrysogaster, AMD), and captive breeding programs are beneficial to the ex-situ conservation and wild population recovery of this species. Meanwhile, the gut microbiota is essential for host health, survival, and environmental adaptation. However, changes in feeding environment and food can affect the composition and function of gut microbiota in musk deer, ultimately impacting their health and adaptation. Therefore, regulating the health status of wild and captive AMD through a non-invasive method that targets gut microbiota is a promising approach. Here, 16S rRNA gene sequencing was employed to reveal the composition and functional variations between wild (N = 23) and captive (N = 25) AMD populations. The results indicated that the gut microbiota of wild AMD exhibited significantly higher alpha diversity (P < 0.001) and greater abundance of the phylum Firmicutes, as well as several dominant genera, including UCG-005, Christensenellaceae R7 group, Monoglobus, Ruminococcus, and Roseburia (P < 0.05), compared to captive AMD. These findings suggest that the wild AMD may possess more effective nutrient absorption and utilization, a more stable intestinal microecology, and better adaption to the complex natural environment. The captive individuals displayed higher metabolic functions with an increased abundance of the phylum Bacteroidetes and certain dominant genera, including Bacteroides, Rikenellaceae RC9 gut group, NK4A214 group, and Alistipes (P < 0.05), which contributed to the metabolic activities of various nutrients. Furthermore, captive AMD showed a higher level of 11 potential opportunistic pathogens and a greater enrichment of disease-related functions compared to wild AMD, indicating that wild musk deer have a lower risk of intestinal diseases and more stable intestinal structure in comparison to captive populations. These findings can serve as a valuable theoretical foundation for promoting the healthy breeding of musk deer and as a guide for evaluating the health of wild-released and reintroduced musk deer in the future. KEY POINTS: • Wild and captive AMD exhibit contrasting gut microbial diversity and certain functions. • With higher diversity, certain bacteria aid wild AMD's adaptation to complex habitats. • Higher potential pathogens and functions increase disease risk in captive AMD.

Impacts of future climate on the distribution of 4 bumblebee species in the Qinghai-Tibet Plateau.

Climate warming threatens dozens of bumblebee species across the world, including alpine areas. In plateaus with vast and continuous high-altitude areas, bumblebees' response to climate warming may be relatively optimistic. To study the species' responses to future climate in the Qinghai-Tibet Plateau, we quantified the suitable areas for 4 local bumblebee indicator species under current and future climate scenarios (Shared Socio-economic Pathway 126, 245, 370, and 585 in 2,100) using MaxEnt models. Suitable areas of indicator species were stacked to obtain the species richness layer. According to the acreage and connectivity of suitable areas and the acreage of the high richness area, a warmed climate will be more suitable for bumblebees' distribution compared to the current climate. The SSP 126 and SSP 245 scenario will be the 2 most suitable. Meanwhile, with climate warming, suitable areas and the high richness areas will move to high altitudes and their altitude range will decrease. The greater suitability in warmed climates may be caused by the topography of plateaus, which provides an opportunity for bumblebees to migrate to cooler areas. However, mitigation of warming is still necessary because an excessively warm climate will decrease bumblebees' habitat suitability. In plateaus, species in lower altitudes will migrate to higher altitudes, conservation in high altitudes should pay attention to not only original species, but also immigrated species. In lower altitudes, many species will migrate to higher altitudes, then local bumblebee diversity will decrease. Local conservation should be focused on these lower altitude areas.

Safety and efficacy of peroral endoscopic myotomy with standard myotomy versus short myotomy for treatment-naïve patients with type II achalasia: a prospective randomized trial.

BACKGROUND AND AIMS: Peroral endoscopic myotomy (POEM) has emerged as an effective endoscopic treatment modality for achalasia. However, there is no consensus regarding the length of muscle bundle dissection during POEM. The most commonly used approach is standard myotomy (about 10 cm). We herein compared the outcomes between standard myotomy versus short myotomy for the management of treatment-naïve patients with type II achalasia. METHODS: This was a prospective, single-center, randomized trial in China. Previously untreated adults with a clinical diagnosis of type II achalasia, confirmed by manometric testing, were enrolled between February 2018 and February 2019. Patients were randomly assigned to POEM with standard myotomy or short myotomy. Clinical data on demographic characteristics, operative parameters, pre- and postoperative Eckardt scores, esophageal manometry results, 24-hour pH test, and adverse events were recorded and compared between the 2 groups. RESULTS: Of 100 randomized patients, 94 underwent treatment (48 in the standard myotomy group and 46 in the short myotomy group), and 91 (97%) completed the study. POEM was successfully accomplished in most patients (97.8%). The primary outcome of treatment success occurred in 45 of 48 patients (93.8%) in the standard myotomy group versus 44 of 46 (95.7%) in the short myotomy group, with no statistically significant difference between the 2 groups (P = .520). There were no significant between-group differences in postoperative esophageal manometry, Eckardt score, diameter of the esophageal lumen, quality of life, procedure-related adverse events, or reflux esophagitis (P > .05). Postoperative abnormal esophageal acid exposure occurred more often in the standard myotomy group than in the short myotomy group (21/48 patients [43.8%] vs 11/46 patients [23.9%], P = .042). Meanwhile, the short myotomy group showed a significant reduction in total procedure time compared with the standard myotomy group (31.2 ± 15.3 minutes vs 45.6 ± 16.2 minutes, respectively, P < .05). CONCLUSIONS: Among treatment-naïve patients with type II achalasia, standard and short POEM were comparable in terms of providing treatment efficacy and improving quality of life at 1 year, whereas short POEM is technically simpler to perform and requires less procedure time. Moreover, the short POEM approach resulted in fewer cases of postoperative abnormal esophageal acid exposure. (Clinical trial registration number: ChiCTR1800014989.).

A novel large deletion in the APC gene associated with Gardner syndrome in a Chinese family.

INTRODUCTION: Gardner syndrome is a hereditary disease characterized by familial adenomatous polyposis (FAP), accompanied by soft tissue tumors. MATERIAL AND METHODS: a Chinese FAP family was enrolled and followed-up for three years. RESULTS: a novel large germline fragment deletion (EX10_16DEL) of the adenomatous polyposis coli (APC) gene was identified by multiplex ligation-dependent probe amplification (MLPA). An unexpected abdominal tumor grew two years after a subtotal colectomy of the proband. The immunohistochemistry study of the abdominal tumor showed SMA(focal+), calponin(+), ß-catenin(nucleus+) and CD34(focal+), CD117(-), which was consistent with a desmoid tumor. CONCLUSION: when a FAP related desmoid tumor appears, the possibility of Gardner syndrome should be considered. This is the first largest deletion of the APC gene in the Chinese population associated with Gardner syndrome.

[Actions of endogenous hydrogen sulfide on colonic hypermotility in a rat model of chronic stress].

The present study was designed to investigate the potential role of endogenous hydrogen sulfide (H2S) in chronic stress-induced colonic hypermotility. Male Wistar rats were submitted daily to 1 h of water avoidance stress (WAS) or sham WAS (SWAS) for 10 consecutive days. The total number of fecal pellets was counted at the end of each 1 h of WAS or SWAS session. Organ bath recordings were used to test the colonic motility. H2S production of colon was determined, and immunohistochemistry and Western blot were performed on rat colonic samples to detect the distribution and expression of H2S-producing enzymes. The results showed that i) repeated WAS increased the number of fecal pellets per hour and the area under the curve (AUC) of the spontaneous contractions of colonic strips (P < 0.05), ii) repeated WAS decreased the endogenous production of H2S and the expression of H2S-producing enzymes in the colon devoid of mucosa and submucosa (P < 0.001), iii) cystathionine-γ-lyase (CSE) was strongly expressed in the cytosols of the circular and longitudinal smooth muscle cells and the nucleus of the myenteric plexus neurons, iv) cystathionine-ß-synthase (CBS) was primarily localized in the cytosols of myenteric plexus neurons and weakly localized in the epithelial cells and v) inhibitors of H2S-producing enzymes increased the contractile activity of colonic strips in the SWAS rats (P < 0.001). In conclusion, the results suggest that the colonic hypermotility induced by repeated WAS may be associated with the decreased production of endogenous H2S.

[Effects of methane on proximal colon motility of rats and ion channel mechanisms].

OBJECTIVE: To explore the effects of methane on proximal colon motility and elucidate its ion channel mechanisms. METHODS: The circular muscle and longitudinal muscle strips of proximal colon were isolated from rats. An isometric force transducer and a biology signal collection system were employed to observe the effects of methane on spontaneous contractile activity of muscle strips. The effects of methane on longitudinal muscle strips were observed after pre-treatment of these strips with tetrodotoxin (TTX) or N-nitro-L-arginine methylester (L-NAME). Single cell of colonic smooth muscle was isolated by collagenase and then whole-cell patch clamp technique was used to record voltage dependent potassium current (IKV) and large conductance Ca(2+)-activated K(+) current (IBKca) in the absence or presence of methane. RESULTS: Methane significantly attenuated the contractile amplitude of longitudinal muscle strips (from (1.12 ± 0.27) to (0.99 ± 0.31) g, n = 19, P = 0.013) whereas there were no changes in some longitudinal muscle strips (n = 6) in the presence of methane. The inhibitory effect of methane persisted after the pre-treatment of longitudinal muscle strips with TTX or L-NAME. Methane had no effects on the contractile cycle time of longitudinal muscle strips and contractile activity of circular muscle strips. And 3% methane solution significantly increased the density of IKV (from (13.3 ± 1.0) pA/pF to (18.5 ± 1.4) pA/pF, at +60 mV, n = 11, P = 0.001) versus the control group whereas methane had no effect on IBKca (all P > 0.05). CONCLUSION: Methane can inhibit contractile activity of proximal colonic longitudinal muscle by activating voltage dependent potassium channel and increasing IKV.

Analysis of complete mitochondrial genome sequence of Pontania dolichura (Hymenoptera: Tenthredinidae) (Thomson, 1871).

Pontania dolichura is a leaf-eating pest that mainly damages willow trees and is widely distributed in northern regions. In this study, we sequenced the entire mitochondrial genome of P. dolichura (GenBank accession number: MZ726800). The circular gene was 16,104 bp in length and comprised 38 column elements, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and a non-coding control region. Most of the PCGs of P. dolichura have typical ATN (Met) start codons and typical TAN stop codons. The A + T contents of the genome, PCGs, transfer RNAs (tRNAs), and ribosomal RNAs (rRNAs) were 80.32%, 78.66%, 81.94%, and 82.59%, respectively. Phylogenetic analysis supported the close genetic relationship between P. dolichura and Mesoneura rufonota indicating that the two species share more recent common ancestor gene. These data will be useful for further molecular identification and population genetics studies.

Transcriptomic Analyses Suggest the Adaptation of Bumblebees to High Altitudes.

Determining the adaptive mechanisms by which bumblebees adapt to high altitudes can help us to better understand their distribution, providing a basis for the future protection and utilization of bumblebee resources. For this study, the adaptive mechanisms of two dominant bumblebee species in the northeastern Qinghai-Tibet Plateau-Bombus kashmirensis and B. waltoni-were studied through transcriptomics methods. For each species, enrichment analysis of the differentially expressed genes and gene set enrichment analysis were carried out between samples collected at different altitudes (4000 m, 4500 m, and 5000 m). The results indicate that these bumblebees tend to up-regulate energy metabolism-related genes when facing extremely high-altitude environments. Of the enriched pathways up-regulated in higher altitudes, the pentose and glucuronate interconversions pathway presented the most severe up-regulation in multiple comparisons of different altitudes for B. kashmirensis, as well as the AMPK signaling pathway, which was found to be up-regulated in both species. Notably, limited by the extreme hypoxic conditions in this study, oxidative phosphorylation was found to be down-regulated with increasing altitude, which is uncommon in studies on bumblebee adaptation to high altitudes.

[Effects of cholecystokinin octapeptide on the contractile activity of guinea-pig colonic smooth muscles, L-type calcium currents and membrane potentials of myocytes].

OBJECTIVE: To investigate the effects and mechanism of cholecystokinin octapeptide (CCK-8S) on the contractile activity of smooth muscles, L-type calcium current and membrane potentials of proximal colon myocytes in guinea pig. METHODS: (1) Strips of proximal colon were obtained from adult guinea pigs. The contraction of these stripes was measured by a RM6240 multi-channel physiological signal system. (2) Suspension of single smooth muscle cells (SMCs) were obtained from proximal colon and isolated by enzymatic digestion. The effect of CCK-8S on intracellular calcium concentration ([Ca(2+)]i) of SMCs was examined by fura-2-loaded microfluorimetric measurement. (3) Resting potential (RP), action potential (AP) and L-type calcium current (I(Ca-L)) were recorded by patch-clamp technique. RESULTS: (1) The contractile amplitude and frequency of muscle stripes enhanced by CCK-8S (10(-7) mol/L) were (149 ± 12)% and (132 ± 13)% respectively of those of control group (all P < 0.05). They were significantly attenuated by pretreating strips with CCK1 receptor antagonist devazepide (10(-7) mol/L), L-type calcium channel blocker nifedipine (10(-5) mol/L), Ca(2+)-ATPase inhibitor TG (thapsigargin) (10(-5) mol/L) and BA (boric acid) (10(-5) mol/L) respectively. (2) [Ca(2+)]i of SMCs intensified by CCK-8S was (738 ± 24)% of that of control group. And it was inhibited by pretreating SMCs with devazepide (all P < 0.05). (3) After the superfusion of CCK-8S, RP depolarized to (52 ± 9)%, the exogenously stimulated peak values of AP rose to (140 ± 4)% and fast repolarization time of AP decreased to (61 ± 13)% (all P < 0.05). They were significantly inhibited when these cells were pretreated with devazepide and/or nifedipine (n = 8, P < 0.05 for each group) whereas CI 988 had little effect. (4) The CCK-8S-evoked I(Ca-L) of SMCs at the voltage of + 10 mV was boosted to (138 ± 7)%. Such an effect was suppressed by a pretreatment with nifedipine, devazepide, TG and BA respectively. In the presence of an inhibitor of inositol 1,4,5-trisphosphate (IP3) receptors, heparin (10(-6) mol/L) and an protein kinase C (PKC) inhibitor, saurosporine (10(-6) mol/L), CCK-8S did not significantly intensify I(Ca-L) (all P > 0.05). CONCLUSION: CCK-8S promotes proximal colon contraction by CCK1 receptors through the activation of IP3-mediated PKC pathway.

Characteristics of complete chloroplast genome of a high-quality forage on Qinghai-Tibet Plateau, Medicago archiducis-nicolai Sirj. (Fabaceae：Trifolieae).

Medicago archiducis-nicolai Sirj. is a well-known high-quality forage as its good palatability and strong tolerance to drought, cold and saline-alkali stress. Here, the complete chloroplast genome sequence of M. archiducis-nicolai was reported. The size of the complete chloroplast genome is 127,072 bp in length. The chloroplast genome has no inverted repeat (IR) regions, which is very common in the family Fabaceae. The M. archiducis-nicolai chloroplast genome encodes 106 genes: 72 protein-coding genes, 30 tRNAs, and 4 rRNAs. The phylogenetic analysis result strongly suggested that M. archiducis-nicolai is a distinct lineage in Medicago, being sister to highly supported clade composed of three species (M. hybrida, M. papillosa and M. sativa).

Complete chloroplast genome of a high-quality forage in north China, Medicago ruthenica (Fabaceae：Trifolieae).

Medicago ruthenica is a well-known high-quality forage due to its good palatability and strong tolerance to drought, cold and saline-alkali stress. Here, the complete chloroplast genome sequence of M. ruthenica was reported. The chloroplast genome is 126,939 bp in length. This chloroplast genome has no inverted repeat (IR) regions, which is very common in the family Fabaceae. The M. ruthenica chloroplast genome encodes 107 genes, including 73 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Phylogenetic analysis result strongly suggested that M. ruthenica is a distinct lineage in Medicago, being sister to highly supported clade composed of three species (M. hybrida, M. papillosa and M. sativa).

The complete chloroplast genome sequence of Stellera chamaejasme f. chrysantha (Thymelaeaceae).

Stellera chamaejasme L. f. chrysantha S. C. Huang is a toxic perennial herb of Thymelaeaceae and has the potential for medicine as Stellera chamaejasme L. Here, we present the complete chloroplast genome sequence of S. chamaejasme f. chrysantha based on Illumina sequencing data. The complete chloroplast genome sequence is 173,364 bp in size and contains four subregions: a pair of inverted repeats (IRs, each for 41,978 bp), a large single-copy region (LSC, 86,558 bp), and a small single-copy region (SSC, 2,850 bp). 141 genes were recognized in the assembled sequence: 93 protein-coding genes (PCGs), 38 tRNAs, 8 rRNAs, and 2 pseudo genes. The phylogenetic analysis result strongly supported that Stellera chamaejasme f. chrysantha was closely related to S. chamaejasme L.

Actions of hydrogen sulfide and ATP-sensitive potassium channels on colonic hypermotility in a rat model of chronic stress.

OBJECTIVE: To investigate the potential role of hydrogen sulphide (H(2)S) and ATP-sensitive potassium (K(ATP)) channels in chronic stress-induced colonic hypermotility. METHODS: Male Wistar rats were submitted daily to 1 h of water avoidance stress (WAS) or sham WAS (SWAS) for 10 consecutive days. Organ bath recordings, H(2)S production, immunohistochemistry and western blotting were performed on rat colonic samples to investigate the role of endogenous H(2)S in repeated WAS-induced hypermotility. Organ bath recordings and western blotting were used to detect the role of K(ATP) channels in repeated WAS. RESULTS: Repeated WAS increased the number of fecal pellets per hour and the area under the curve of the spontaneous contractions of colonic strips, and decreased the endogenous production of H(2)S and the expression of H(2)S-producing enzymes in the colon devoid of mucosa and submucosa. Inhibitors of H(2)S-producing enzymes increased the contractile activity of colonic strips in the SWAS rats. NaHS concentration-dependently inhibited the spontaneous contractions of the strips and the NaHS IC(50) for the WAS rats was significantly lower than that for the SWAS rats. The inhibitory effect of NaHS was significantly reduced by glybenclamide. Repeated WAS treatment resulted in up-regulation of Kir6.1 and SUR2B of K(ATP) channels in the colon devoid of mucosa and submucosa. CONCLUSION: The colonic hypermotility induced by repeated WAS may be associated with the decreased production of endogenous H(2)S. The increased expression of the subunits of K(ATP) channels in colonic smooth muscle cells may be a defensive response to repeated WAS. H(2)S donor may have potential clinical utility in treating chronic stress-induced colonic hypermotility.