Identification of the Xyloglucan Endotransglycosylase/Hydrolase (XTH) Gene Family Members Expressed in Boehmeria nivea in Response to Cadmium Stress.

Xyloglucan endotransglycosylase/hydrolase (XTH) genes play an important role in plant resistance to abiotic stress. However, systematic studies of the response of Boehmeria nivea (ramie) XTH genes (BnXTHs) to cadmium (Cd) stress are lacking. We sought to identify the BnXTH-family genes in ramie through bioinformatics analyses and to investigate their responses to Cd stress. We identified 19 members of the BnXTH gene family from the ramie genome, referred to as BnXTH1-19, among which BnXTH18 and BnXTH19 were located on no chromosomes and the remaining genes were unevenly distributed across 11 chromosomes. The 19 members were divided into four groups, Groups I/II/IIIA/IIIB, according to their phylogenetic relationships, and these groups were supported by analyses of intron-exon structure and conserved motif composition. A highly conserved catalytic site (HDEIDFEFLG) was observed in all BnXTH proteins. Additionally, three gene pairs (BnXTH6-BnXTH16, BnXTH8-BnXTH9, and BnXTH17-BnXTH18) were obtained with a fragment and tandem-repeat event analysis of the ramie genome. An analysis of cisregulatory elements revealed that BnXTH expression might be regulated by multiple hormones and abiotic and biotic stress responses. In particular, 17 cisregulatory elements related to abiotic and biotic stress responses and 11 cisregulatory elements related to hormone responses were identified. We also found that most BnXTH genes responded to Cd stress, and BnXTH1, BnXTH3, BnXTH6, and BnXTH15 were most likely to contribute to the Cd tolerance of ramie, as evidenced by the substantial increases in expression under Cd treatment. Heterologous expression of BnXTH1, BnXTH6, and BnXTH15 significantly enhanced the Cd tolerance of transgenic yeast cells. These results suggest that the BnXTH gene family is involved in Cd stress responses, laying a theoretical foundation for functional studies of BnXTH genes and the innovative breeding of Cd-tolerant ramie.

Hydrophobic Modification on the Surface of SiO2 Nanoparticle: Wettability Control.

Good control of the morphology, particle size, and wettability of silica nanoparticles is of increasing importance to their use in a variety of fields. Here, we propose a strategy to tune the surface wettability of nanosilica by changing the dosage of a chemical modifier. A series of measurements, including scanning electron microscopy (SEM), laser scatting technique, Fourier transform infrared (FTIR) spectroscopy, thermogravimetry, and surface hydroxyl number and water contact angle measurement, were conducted to verify the surface chemistry and wettability of these nanoparticles. Through controlled chemical modification, the contact angle of the treated nanoparticles increases from 34.7 to 155° with increasing amount of dichlorodimethylsilane (DCDMS) within a molar ratio (MR) between DCDMS and nanoparticles of 5.17. The number of hydroxyl groups covered on the particle surface decreases gradually from 1.79 to 0.47, and the surface grafting rate could reach 73.7%. As the addition of dichlorodimethylsilane equals MR 5.17, the contact angle reaches the maximum value of 155°, which displays excellent superhydrophobicity. After surpassing the point of MR 5.17, the contact angle does not increase but starts to decrease, ultimately remaining stable at 135°. It can be concluded that the surface wettability of nano-SiO2 particles can be precisely modulated by varying the amounts of the modifier. Furthermore, the modulating mechanism of the process occurring on the surface of SiO2 particles has been investigated at the molecular level.

Divergent Expression Patterns and Function of Two cxcr4 Paralogs in Hermaphroditic Epinephelus coioides.

Chemokine receptor Cxcr4 evolved two paralogs in the teleost lineage. However, cxcr4a and cxcr4b have been characterized only in a few species. In this study, we identified two cxcr4 paralogs from the orange-spotted grouper, Epinephelus coioides. The phylogenetic relationship and gene structure and synteny suggest that the duplicated cxcr4a/b should result from the teleost-specific genome duplication (Ts3R). The teleost cxcr4 gene clusters in two paralogous chromosomes exhibit a complementary gene loss/retention pattern. Ec_cxcr4a and Ec_cxcr4b show differential and biased expression patterns in grouper adult tissue, gonads, and embryos at different stages. During embryogenesis, Ec_cxcr4a/b are abundantly transcribed from the neurula stage and mainly expressed in the neural plate and sensory organs, indicating their roles in neurogenesis. Ec_Cxcr4a and Ec_Cxcr4b possess different chemotactic migratory abilities from the human SDF-1α, Ec_Cxcl12a, and Ec_Cxcl12b. Moreover, we uncovered the N-terminus and TM5 domain as the key elements for specific ligand⁻receptor recognition of Ec_Cxcr4a-Ec_Cxcl12b and Ec_Cxcr4b-Ec_Cxcl12a. Based on the biased and divergent expression patterns of Eccxcr4a/b, and specific ligand⁻receptor recognition of Ec_Cxcl12a/b⁻Ec_Cxcr4b/a, the current study provides a paradigm of sub-functionalization of two teleost paralogs after Ts3R.

Divergent Expression Patterns and Function Implications of Four nanos Genes in a Hermaphroditic Fish, Epinephelus coioides.

Multiple nanos genes have been characterized in several fishes, but the functional implications of their various expression patterns remain unclear. In this study, we identified and characterized four nanos genes from a hermaphroditic fish orange-spotted grouper, Epinephelus coioides. Ecnanos1a and Ecnanos1b show divergent expression patterns, and the dynamic expression change of Ecnanos1a in pituitaries during sex change is associated with testis differentiation and spermatogenesis. Ecnanos2 and Ecnanos3 might be germline stem cells (GSCs) and primordial germ cells (PGCs)-specific markers, respectively. Significantly, Ecnanos3 3'-untranslated region (UTR) is necessary for PGC specific expression, where a non-canonical "GCACGTTT" sequence is required for miR-430-mediated repression of Ecnanos3 RNA. Furthermore, grouper Dead end (Dnd) can relieve miR-430 repression in PGCs by associating with a 23 bp U-rich region (URR) in Ecnanos3 3'-UTR. The current study revealed the functional association of multiple nanos genes with PGC formation and germ cell development in orange-spotted grouper, and opened up new possibilities for developing biotechnologies through utilizing the associations between Ecnanos3 and PGCs or between Ecnanos2 and GSCs in the hermaphroditic fish.

Genetic mechanism of body size variation in groupers: Insights from phylotranscriptomics.

Animal body size variation is of particular interest in evolutionary biology, but the genetic basis remains largely unknown. Previous studies have shown the presence of two parallel evolutionary genetic clusters within the fish genus Epinephelus with evident divergence in body size, providing an excellent opportunity to investigate the genetic basis of body size variation in vertebrates. Herein, we performed phylotranscriptomic analysis and reconstructed the phylogeny of 13 epinephelids originating from the South China Sea. Two genetic clades with an estimated divergence time of approximately 15.4 million years ago were correlated with large and small body size, respectively. A total of 180 rapidly evolving genes and two positively selected genes were identified between the two groups. Functional enrichment analyses of these candidate genes revealed distinct enrichment categories between the two groups. These pathways and genes may play important roles in body size variation in groupers through complex regulatory networks. Based on our results, we speculate that the ancestors of the two divergent groups of groupers may have adapted to different environments through habitat selection, leading to genetic variations in metabolic patterns, organ development, and lifespan, resulting in body size divergence between the two locally adapted populations. These findings provide important insights into the genetic mechanisms underlying body size variation in groupers and species differentiation.

Oxford Nanopore Technologies (ONT) Full-length Transcriptomics Shows Electroacupuncture ameliorates Lipid Metabolic Disorder through Suppressing Hepatic Pdia3/Perk/Qrich1 Expression in Rats.

BACKGROUND: The incidence of dyslipidemia increases after menopause. Electroacupuncture (EA) has been recommended for menopause-related disease. However, the positive effect on lipid metabolism disorders is still unclear. OBJECTIVES: To investigate the underlying mechanism of EA treatment on lipid metabolism disorders through ONT full-length transcriptome sequencing Methods: Adult female SD rats were randomly divided into Ctrl, sham operation+high-fat feed(Sham+HFD), Ovariectomized+high-fat feed (OVX+HFD), Ovariectomized+high-fat feed + Atorvastatin (OVX+HFD+ATO) and OVX+HFD+EA groups. Periovarian adipose tissue around the bilateral ovaries of rats in the Sham+HFD group was resected. Rats in the OVX+HFD, OVX+HFD+ATO and OVX+HFD+EA groups were subjected to bilateral oophorectomy to prepare the ovariectomized rat model. Treatment was applied to rats in the OVX+HFD+EA group. ST36, PC6, SP6, BL18 and ST40 were the selected acupoints. Daily food intake and body weights of rats were recorded. The samples were collected 30 days after treatment. The serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein (HDL-C) were detected to assess the improvement of lipid metabolism disorders. HE and oil red O staining were used to stain the liver tissues. Total RNA was extracted from liver tissues, and its transcriptional changes were determined by high-throughput sequencing. Additionally, RTÁqPCR and immunofluorescence staining were used to verify the crucial signal pathway screened by the ONT fullÁlength transcriptome sequencing. RESULTS: EA treatment resulted in a lowered weight of perirenal fat and liver and a significant improvement in the color of the liver. In addition, EA could improve the lipid profile and hepatic steatosis in OVX+HFD rats. According to fullÁlength transcriptome sequencing, 2292 genes showed differential expression in the OVX+HFD group; of these, 1121 were upregulated and 1171 down-regulated. 609 DEGs were found in the OVX+HFD+EA group compared to the OVX+HFD group; 235 up-regulated and 374 down-regulated. We also found that 77 genes are significantly upregulated after EA intervention through Venn map analysis (including Agtr1a, Pdia3, etc.), which may be the targeted genes for EA treatment of lipid metabolism disorders. Finally, we verified the expression of Pdia3, Perk and Qrich1 levels in liver tissues. HFD feeding could increase the expression of Pdia3 and its downstream signal pathways molecular Perk and Qrich1. But these effects were reversed by EA treatment, the results demonstrated that the expression of pdia3, Perk, as well as Qrich1 of OVX+HFD rats had a decreasing trend after EA treatment. CONCLUSIONS: EA could ameliorate lipid metabolic disorder in OVX+HFD rats. The Pdia3/Perk/Qrich1 signal pathway may play crucial roles in the improvement of lipid metabolism disorder of OVX+HFD rats after EA treatment.

Orange-spotted grouper (Epinephelus coioides) toll-like receptor 22: molecular characterization, expression pattern and pertinent signaling pathways.

The toll-like receptors (TLRs) are an important gene family in host innate immunologic surveillance. The TLR22 gene is an essential member of the TLRs that is only found in aquatic animals and has been detected in some bony fish. Here, a TLR22 homolog, EcTLR22, was characterized in the orange-spotted grouper (Epinephelus coioides) via homology cloning. The 3321 bp full-length cDNA sequence of EcTLR22 was obtained, which included an open reading frame of 2880 bp encoding a putative peptide of 960 amino acids containing three highly typical domains with the characteristics of TLR family members. The deduced amino acid sequence of EcTLR22 showed a relatively high similarity to flounder TLR22. Phylogenetic analysis showed that the orange-spotted grouper TLR22 sequence was clustered with those of Perciforme, such as flounder and croaker. Real-time quantitative PCR analysis revealed broad expression of EcTLR22, with relatively high expression detected in the head kidney, trunk kidney, spleen, peripheral blood leukocytes (PBLs) and heart of orange-spotted grouper. After injection with Vibrio alginolyticus, there was significant up-regulation of the expression of EcTLR22 in the spleen. In evaluating unstimulated/stimulated head kidney leukocytes and spleen leukocytes, a significant increase in EcTLR22 mRNA expression was detected, which implied a sensitive immune response. Furthermore, four important molecules for signal transduction, MyD88, TRIF, TNF-α and IRF3, were chosen to analyze the role of the EcTLR22 signaling pathway in anti-pathogen responses. Upon LPS or Poly I:C challenge, expression of the four genes was induced, with an increasing tendency detected in head kidney leukocytes, suggesting that the four genes might work with EcTLR22 in host defense against pathogenic microbes.

Differential expression analysis of nuclear oligomerization domain proteins NOD1 and NOD2 in orange-spotted grouper (Epinephelus coioides).

Nucleotide-binding oligomerization domain-containing proteins-1 and -2 (NOD1 and NOD2) are members of the NOD-like receptors (NLRs) family. They are both cytoplasmic receptors, and sense microbial infections/danger molecules to induce host innate immune response. In this study, the full-length ORF sequences of NOD1 and NOD2 were cloned, and the putative amino acid sequences were identified in orange-spotted grouper (Epinephelus coioides). The complete open reading frame (ORF) of grouper NOD1 contained 2823 bp encoding a 940 amino acid protein. Grouper NOD2 cDNA contained a 2967 bp ORF, encoding a protein of 988 amino acid residues. Both grouper NOD1 and NOD2 had similar domains to human and fish counterparts. Phylogenetic tree analysis showed that grouper NOD1 clustered with grass carp, zebrafish and channel catfish, while NOD2 was most closely related to fugu. Expression patterns of grouper NOD1 and NOD2 were next studied. NOD1 had the highest level of expression in skin while NOD2 in trunk kidney. Post Vibrio alginolyticus (strain EcGS020401), lipopolysaccharide (LPS) or PolyI:C challenges, gene expression of grouper NOD1 and NOD2 was stimulated to different extents. NOD1 showed a significant enhancement after LPS stimulation, but NOD2 increased more significantly after PolyI:C invasion, indicating that NOD1 and NOD2 may exert different effects on the eradication of bacteria and virus. The adaptor protein RIP-like-interacting CLARP kinase (RICK) and downstream molecule interleukin-8 (IL-8) were also induced at different levels after stimulation, which indicated that NOD1 and NOD2 signal transduction was involved in grouper innate immune protection against bacterial and viral infections.

Exploring the effect of metal ions and counteranions on the structure and magnetic properties of five dodecanuclear Co(II) and Ni(II) clusters.

We present the synthesis, characterization of the structures, and magnetic properties of five isostructural dodecanuclear coordination clusters of Ni(II) and Co(II): [Co(12)(bm)(12)(NO(3))(O(2)CMe)(6)(EtOH)(6)](NO(3))(5) (1), [Ni(12)(bm)(12)(NO(3))(O(2)CMe)(6)(H(2)O)(3)(EtOH)(3)](NO(3))(5)·2H(2)O (2), mixed-metal composition (Ni/Co 1:1) [Co(6)Ni(6)(bm)(12)(NO(3))(O(2)CMe)(6)(NO(3))(5) (3), and [M(12)(bm)(12)(NO(3))(O(2)CMe)(6)(EtOH)(6)](ClO(4))(5) (M=Co (4), Ni (5)), in which Hbm=(1H-benzimidazol-2-yl)methanol. They consist of analogous structural cores that are constructed by three cubanes (M(4)O(4)) that surround the templating nitrate and bridging auxiliary acetate and the directing ligands bm. They have different magnetic behaviors. Whereas there is the absence of the out-of-phase ac susceptibility (χ'') for the Ni(II)-based compounds 2 and 5, the Co(II)-containing compounds 1, 3, and 4 have prominent χ'' signals that exhibit frequency dependence, which indicates slow magnetic relaxation behavior above 1.8 K. In particular, the larger perchlorate counterions in 4 further change the overall correlation interaction between clusters, thus leading to an enhanced blocking temperature for the less-symmetrical 4 (pseudo-C(3)) relative to 1 and 3 (true C(3)). Interestingly, electrospray ionization mass spectrometry (ESI-MS) indicates that the three dodecanuclear clusters of 1-3 retain their compositions in solution. The mixed-metal cluster cores of 3 are formed based on the nature of the interchangeability between metal centers in solution.

[Expression and significance of HOXA10 gene in the eutopic and ectopic endometrium of endometriosis].

OBJECTIVES: To investigate the expression and clinical significance of HOXA10 gene in the eutopic and ectopic endometrium of endometriosis. Mehtods Between Jan.2009 to Aug.2010, 30 patients with endometriosis undergoing laparoscopic surgery in Maternal and Children's Hospital of Foshan. Eutopic and ectopic endometrium were obtained. In the mean time, 30 patients with benign ovary cyst or tubal infertility undergoing laparoscopic surgery were selected as controls. Their uterine endometrium were obtained real-time fluorescent quantitation, western blot and immunohistochemistry technique were used to detect mRNA and protein expression of HOXA10 gene in the eutopic endometrium group, ectopic endometrium group and control group. RESULTS: The mRNA and protein expression of HOXA10 gene were 0.61 ± 0.07 and 0.47 ± 0.05 in the eutopic endometrium of endometriosis, 0.64 ± 0.06 and 0.50 ± 0.05 in ectopic endometrium of endometriosis, which were significantly lower than 1.22 ± 0.14 and 1.42 ± 0.14 in control group (P < 0.01). However, the mRNA and protein expression of HOXA 10 between eutopic and ectopic endometrium of endometriosis did not reach statistical difference (P > 0.05). The expression of HOXA10 in eutopic and ectopic endometrium of endometriosis were decreased by immunohistochemistry staining. CONCLUSION: The lower expression of HOXA10 gene in the eutopic and ectopic endometrium of endometriosis might be associated with pathogenesis and infertility of endometriosis.

Impact of different stereoisomers of inositol on insulin sensitivity of gestational diabetes mellitus patients.

BACKGROUND: Inositol is a hexa-carbon polyol, a naturally soluble vitamin, often found in various foods. AIM: To discuss the impact of different stereoisomers of inositol on insulin sensitivity of gestational diabetes mellitus (GDM) patients. METHODS: Eighty GDM pregnant women were divided into four groups according to their treatment received: A group (placebo folic acid 400 µg/d), B group [myo-inositol (MI) 1500 mg, twice a day], C group [D-chiro-inositol (DCI) 250 mg, twice a day], and D group (inositol MI and inositol DCI 1500 mg/250 mg, twice a day). Each patient routinely used dietary guidance adjustments and did some safe and effective aerobic exercise in addition to receiving placebo or inositol from GDM diagnosis to delivery. Triglyceride, total cholesterol, fasting plasma glucose, oral glucose tolerance test postprandial glucose (2 h postprandial blood glucose), fasting insulin, fasting plasma glucose, and glycosylated hemoglobin levels and Homeostasis Model Assessment-insulin resistance (HOMA-IR) and Homeostasis Model Assessment-insulin sensitivity index (HOMA-ISI) scores were determined before treatment and 8 wk after treatment onset. Adverse maternal and infant outcomes, including hypoglycemia, excessive amniotic fluid, premature infants, macrosomia, fetal distress etc., were also recorded. RESULTS: There was no statistical difference in the baseline data of each group. The levels of 2 h blood glucose, glycosylated hemoglobin, fasting insulin, total cholesterol, and triglyceride in the B, C, and D groups were significantly lower than those in the control group (A group) after treatment (P < 0.05). Moreover, compared with the B group, the level of the above indexes in the C and D groups decreased more significantly, and the differences were statistically significant (P < 0.05). The HOMA-IR of B, C, and D groups decreased significantly, and the HOMA-ISI increased significantly compared with the A group, and the differences were statistically significant (P < 0.05), among which the decrease of HOMA-IR and the increase of HOMA-ISI were more significant in the C and D group compared with the B group (P < 0.05). The occurrence rate of adverse maternal and infant outcomes in the C and D group was significantly lower than that in the control group (A group), and the differences were statistically significant (P < 0.05). CONCLUSION: Treatment with different inositol stereoisomers (inositol MI and inositol DCI) can improve insulin sensitivity and reduce insulin resistance in diabetic patients, and inositol DCI has a better curative effect than inositol MI.

[Total Prolift(TM) system surgery for the repair of recurrent severe pelvic organ prolapse].

OBJECTIVE: To evaluate the clinical outcomes of total Prolift(TM) system surgery for the repair of recurrent severe pelvic organ prolapse. METHODS: The clinical records of 13 cases of recurrent severe pelvic organ prolapse were retrospectively analyzed. The characteristic was vaginal vault prolapse in combination with anterior and posterior wall prolapse. Anatomic effect (according to POP-Q score), functional effect (according to prolapse quality of life) and sexual life (according to sexual life score) of each patient were assessed. All subjects were post-menopausal with a mean age (60 ± 9) years and a mean BMI of (25.6 ± 2.9) kg/m(2). RESULTS: All operations were accomplished successfully without any impairment of bladder, urethra, rectum, great vessels or nerves or blood transfusion. The mean operation time was (61 ± 18) minutes and the mean blood loss (155 ± 84) ml. All patients were followed up for a medium of 19 (15 - 30) months. Except for one post-operative urinary retention for a period of 7 days, 12 patients were able to micturate spontaneously 1 - 2 d post-operatively with a residual urine volume of less than 100 ml. During follow-up, mesh erosion (n = 1), de novo overactive bladder (n = 1), one algopareunia (n = 1) and dyspareunia (n = 1) occurred. Postoperative Quality of Life Scores improved significantly (P < 0.01). However, sexual life scores was impaired (P < 0.05). The anatomic cure rate was 92.3% and the patient subjective satisfactory rate 84.6%. CONCLUSION: The total Prolift(TM) system surgery represents a safe, simple and useful treatment for recurrent severe pelvic organ prolapse with satisfactory objective clinical outcomes. However, the level of patient subjective satisfaction is lower than objective cure rate. Meanwhile, harmful effects on sexual life remain a main concern.

Whole-genome sequencing of leopard coral grouper ( Plectropomus leopardus) and exploration of regulation mechanism of skin color and adaptive evolution.

Leopard coral groupers belong to the Plectropomus genus of the Epinephelidae family and are important fish for coral reef ecosystems and the marine aquaculture industry. To promote future research of this species, a high-quality chromosome-level genome was assembled using PacBio sequencing and Hi-C technology. A 787.06 Mb genome was assembled, with 99.7% (784.57 Mb) of bases anchored to 24 chromosomes. The leopard coral grouper genome size was smaller than that of other groupers, which may be related to its ancient status among grouper species. A total of 22 317 protein-coding genes were predicted. This high-quality genome of the leopard coral grouper is the first genomic resource for Plectropomus and should provide a pivotal genetic foundation for further research. Phylogenetic analysis of the leopard coral grouper and 12 other fish species showed that this fish is closely related to the brown-marbled grouper. Expanded genes in the leopard coral grouper genome were mainly associated with immune response and movement ability, which may be related to the adaptive evolution of this species to its habitat. In addition, we also identified differentially expressed genes (DEGs) associated with carotenoid metabolism between red and brown-colored leopard coral groupers. These genes may play roles in skin color decision by regulating carotenoid content in these groupers.

An epidemiology study of fecal incontinence in adult Chinese women living in urban areas.

BACKGROUND: Fecal incontinence (FI) has been shown to be a common symptom in Western countries; however, there is few researches focusing on its epidemic condition in Chinese women. We conducted this national population-based epidemiology study to estimate the prevalence and risk factors of FI among adult Chinese women living in urban regions. METHODS: This is a subgroup analysis of a national population-based epidemiology study of FI. Total 28,196 adult women from urban regions of six provinces and municipalities participated in this research from 2014 to 2015. They finished the questionnaire under the direction of trained interviewers. FI was defined as accidental leakage of flatus and/or liquid or solid stool at least once in the past. The FI prevalence trend and risk factors were identified by the Cochran-Armitage test, Chi-square test, and multivariable logistic regression. RESULTS: The prevalence of FI in adult females in urban China was 0.43% (95% confidence interval: 0.35%-0.51%). Among women with FI, 42.96%, 82.96%, and 42.22% reported having leakage of solid, liquid stool, and gas, respectively. The overall FI prevalence and the incidence rate of solid stool/liquid stool/gas leakage increased with age. The mean Wexner score was 4.0% and 12.0% FI patients reported Wexner score ≥9. Body mass index ≥24 kg/m, pelvic organ prolapses, chronic constipation, chronic cough, alcohol consumption, physical diseases including chronic bronchitis and cancer, gynecological diseases like gynecological inflammation are risk factors for FI. Vaginal delivery was the risk factor for FI in females with labor history. CONCLUSIONS: FI was not a common symptom in adult Chinese women living in urban areas and there were some potential modifiable risk factors. TRIAL REGISTRATION: Chinses Clinical Trial Registry: ChiCTR-OCS-14004675; http://www.chictr.org.cn/showproj.aspx?proj=4898.

Assessment of myocardial blood perfusion improved by CD151 in a pig myocardial infarction model.

AIM: To appraise the efficacy of CD151-induced myocardial therapeutic angiogenesis in a pig myocardial infarction model. METHODS: CD151 and anti-CD151 were constructed into the recombinant adeno-associated virus (rAAV) vector. All 26 pigs were subjected to coronary artery ligation or no surgery. Eight weeks after coronary artery ligation, the expression of CD151 was measured by Western blot and immunostaining. Capillary density was evaluated using immunostaining for von Willebrand factor (vWF). 13N-labeled NH3 positron emission computed tomography ([13N]NH3PET) was measured to assess regional myocardial perfusion and the defect area. RESULTS: CD151 gene delivery could increase the expression of CD151 at protein level. Over-expression of CD151 increased the density of total capillaries in the ischemic myocardium, significantly improved the blood perfusion and reduced the defect area percentage. CONCLUSION: This study demonstrated that the rAAV-mediated CD151 gene delivery promoted efficient neovascularization and increased the blood perfusion after myocardial infarction in pigs.

Molecular characterization of marbled eel (Anguilla marmorata) gonadotropin subunits and their mRNA expression profiles during artificially induced gonadal development.

Three cDNA sequences encoding the gonadotropin subunits, common glycoprotein alpha subunit (GTHalpha), FSHbeta and LHbeta subunits were isolated from marbled eel. The cDNA of GTHalpha encodes 116 amino acids with a signal peptide of 24 amino acids and a mature peptide of 92 amino acids. The FSHbeta subunit consists of 127 amino acids with a 22 amino acid signal peptide and a 105 amino acid mature peptide, while the LHbeta subunit consists of 140 amino acids with a 24 amino acid signal peptide and a 116 amino acid mature peptide. Comparison of the deduced amino acid sequences of marbled eel GTHalpha, FSHbeta, and LHbeta with that of other fishes shows a high degree of conservation in the number of cysteine residues and potential N-linked glycosylation sites. The mRNA of GTHalpha, FSHbeta and LHbeta were not only detected in pituitary, but also in ovary and testes by RT-PCR. Quantitative realtime PCR analysis revealed that the GTHalpha and LHbeta transcriptional levels in pituitaries of female and male eels gradually increased during the artificially inducing gonadal development, and peaked at late vitellogenic stage and spermiation stage, respectively. FSHbeta mRNA in the pituitaries of female eels maintained a high level at previtellogenic stage, early vitellogenic stage as well as mid-vitellogenic stage but declined sharply at late vitellogenic stage and migratory nucleus stage. In male eels, the mRNA levels of FSHbeta in the pituitaries were higher at early spermatogenesis stage than at both late spermatogenesis stage and spermiation stage. These results suggested that FSH would be in control of initiation and maintenance of gonadal growth and gametogenesis, whereas LH would be involved in the final gonadal maturation and spermiation/ovulation in the tropic eel Anguilla marmorata.

Interleukin-1beta gene in orange-spotted grouper, Epinephelus coioides: molecular cloning, expression, biological activities and signal transduction.

IL-1beta, a key mediator of inflammation, orchestrates a variety of immune responses by initiating gene expression. Herein, we have cloned and sequenced the IL-1beta in orange-spotted grouper (Epinephelus coioides), produced soluble mature recombinant IL-1beta in Escherichia coli, and characterized its biological properties and downstream signal transduction. The grouper IL-1beta cDNA was 1364bp in length, containing an open reading frame of 765bp. The predicted protein of 254 amino acids revealed the presence of the IL-1 family signature motif and the absence of a conventional ICE cut site. Phylogenetically, the grouper IL-1beta clustered closely with those of teleost belonging to Perciformes and apart from those of mammals. The grouper IL-1beta was constitutively expressed in almost all tissues examined, and was augmented in PBL after the addition of LPS or Poly I:C in vitro. The prokaryotically produced rIL-1beta significantly stimulated the proliferation of grouper head kidney cells, and activated gene expression of IL-1beta and COX-2. Moreover, the rIL-1beta-induced IL-1beta and COX-2 expression were reduced by p38 MAPK inhibitor (SB203580) and JNK inhibitor (SP600125), respectively. Taken together, the present study indicated that grouper IL-1beta may have an important role in grouper immune system and activate similar downstream cascades as its mammalian counterparts.

[Arteriogenesis induced by intramyocardial recombinant adeno-associated virus vector encoding human CD151 cDNA gene transfer in swines with coronary artery occlusion].

OBJECTIVE: To investigate the efficacy of CD151 gene delivery in promoting blood perfusion in swines after myocardial infarction. METHODS: Swines received coronary artery ligation and intramyocardial injection with rAAV-CD151, rAAV-anti-CD151 or rAAV-GFP. Eight weeks after vector injection, Western blot, immunostaining and 13N-labeled NH3 PET were performed to detect gene expression and biological effects of various treatments. RESULTS: High level of CD151 protein expression was detected in the rAAV-CD151 group. The capillary density in the rAAV-CD151 group [(83.8 +/- 6.7) n/mm2] was significantly higher than that in the control group [(33.2 +/- 4.5) n/mm2] and rAAV-GFP group [(41.6 +/- 5.6) n/mm2] (all P<0.05); the arteriole density in the rAAV-CD151 group [(16.4 +/- 2.5) n/mm2] was also higher than that in the control group [(6.6 +/- 2.3) n/mm2] and the rAAV-GFP group [(8.4 +/- 1.6) n/mm2] (all P<0.05). However, the lowest capillary density and arteriole density were evidenced in rAAV-anti-CD151 group. Myocardial blood perfusion was significantly increased in rAAV-CD151 group and significantly reduced in rAAV-anti-CD151 group (all P<0.05 vs. control). CONCLUSION: Intramyocardial injection of rAAV-CD151 could enhance the myocardial express of CD151 protein, increase capillary and arteriole densities and improve blood perfusion in swine with myocardial infarction.

Effect of High-Fat Diet on Immature Female Mice and Messenger and Noncoding RNA Expression Profiling in Ovary and White Adipose Tissue.

Obesity is a chronic multifactorial disease prevalent in many areas of the world and is a major cause of morbidity and mortality. In women, obesity increases the risks of both metabolic and reproductive diseases, such as diabetes and infertility. The mechanisms underlying these effects, especially in young women, are largely unknown. To explore these mechanisms, we established a high-fat diet (HFD) model of obesity in immature female mice. Microarray analysis of gene expression in ovaries and white adipose tissue identified a large number of differentially expressed genes (>1.3-fold change) in both tissues. In ovaries of the HFD group, there were 208 differentially expressed messenger RNAs (mRNAs), including 98 upregulated and 110 downregulated, and 295 differentially expressed lncRNAs (long non coding RNAs), including 63 upregulated and 232 downregulated. In white adipose tissue, there were 625 differentially expressed mRNAs, including 220 upregulated and 605 downregulated in the HFD group, and 1595 differentially expressed lncRNAs, including 1320 and 275 downregulated in the HFD group. Our results reveal significant differences between the transcriptomes of the HFD and control groups in both ovaries and white adipose tissue that provide clues to the molecular mechanisms of diet-induced female reproductive dysfunction and metabolic disorders, as well as biomarkers of risk for these disorders.

One bis-indole alkaloid-voacamine from Voacanga africana Stapf: biological activity evaluation of PTP1B in vitro utilizing enzymology method based on SPRi expriment.

One known bis-indole alkaloid-voacamine was isolated from Voacanga africana Stapf and Surface Plasmon Resonance imaging (SPRi) exprement showed that this alkaloid could be combine with Protein Tyrosine Phosphatase1B (PTP1B). Then the PTP1B activity inhibition experiment display that the compound showed an outstanding promoting activity to PTP1B.