Evolutionary divergence of subgenomes in common carp provides insights into speciation and allopolyploid success.

Hybridization and polyploidization have made great contributions to speciation, heterosis, and agricultural production within plants, but there is still limited understanding and utilization in animals. Subgenome structure and expression reorganization and cooperation post hybridization and polyploidization are essential for speciation and allopolyploid success. However, the mechanisms have not yet been comprehensively assessed in animals. Here, we produced a high-fidelity reference genome sequence for common carp, a typical allotetraploid fish species cultured worldwide. This genome enabled in-depth analysis of the evolution of subgenome architecture and expression responses. Most genes were expressed with subgenome biases, with a trend of transition from the expression of subgenome A during the early stages to that of subgenome B during the late stages of embryonic development. While subgenome A evolved more rapidly, subgenome B contributed to a greater level of expression during development and under stressful conditions. Stable dominant patterns for homoeologous gene pairs both during development and under thermal stress suggest a potential fixed heterosis in the allotetraploid genome. Preferentially expressing either copy of a homoeologous gene at higher levels to confer development and response to stress indicates the dominant effect of heterosis. The plasticity of subgenomes and their shifting of dominant expression during early development, and in response to stressful conditions, provide novel insights into the molecular basis of the successful speciation, evolution, and heterosis of the allotetraploid common carp.

[Nuclear magnetic resonance-based metabolomics revealed metabolite profiles participate in intervention effect of refined moxibustion in gastric ulcer model rats].

OBJECTIVE: To investigate the effect of refined moxibustion on expression of gastric mucosal epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF), and changes of metabolite profiles in gastric ulcer (GU) rats, so as to analyze its mechanism underlying improvement of GU. METHODS: Male SD rats were randomized into control, model, acupoint moxibustion groups (n=6 per group). The GU model was induced by cold-restraint stress. The ignited refined moxa was applied to bilateral "Liangmen" (ST21) and "Zusanli" (ST36) for 3 cones/acupoint, once daily for 7 days. Then, we employed 1H NMR-based metabolomics approach to analyze the metabolic profiles of serum and stomach tissue samples. The conventional histopathological changes of the gastric mucosa were observed by H.E. stain and the expressions of EGFR and VEGF in the gastric mucosa were detected by immunohistochemistry. RESULTS: Compared to the control group, the expression levels of EGFR and VEGF were significantly increased in the model group (P<0.01, P<0.05), and further notably up-regulated in the acupoint moxibustion group (P<0.001, P<0.01). Results of H.E. staining showed damage of the folds of gastric mucosa, disordered arrangement of the glands, infiltration of inflammatory cells and unclear structure of gastric mucosa in the model group, which was relatively milder in the acupoint moxibustion group. 1H-NMR technical analysis showed that in comparison with the control group, 11 and 11 metabolites in the stomach extract and plasma were increased, 10 in the gastric tissue and 3 in the plasma were decreased in the GU model group; while in comparison with the model group, 17 differently expressed metabolites in the gastric extract and 10 metabolites in the plasma restored to their levels of control group after the acupoint moxibustion intervention. These metabolites participate in 12 metabolic pathways including glycine, serine and threonine metabolism, glutathione metabolism, glycine metabolism, alanine, aspartic acid and glutamic acid metabolism, purine metabolism, glyoxylic acid and digarboxylic acid metabolism, biosynthesis of aminoacyl-tRNA, amino sugar and nucleotide sugar metabolism, cysteine and methionine metabolism, citrate cycle, pyruvate metabolism, and the mutual conversion of pentose and glucuronate，suggesting their involvement in moxibustion-induced improvement of GU. CONCLUSION: Refined moxibustion at ST21 and ST36 can up-regulate the expression of EGFR and VEGF in the gastric mucosa and lessen gastric mucosal injury, which may be related to its effects in reducing GU-induced metabolic disorders, including sugar, purine, amino acid, and phospholipid metabolism and antioxidant defense system.

Clinical Study of Stellate Ganglion Block Combined with General Anesthesia on Hemodynamics, Cognitive Function, and Gastrointestinal Function in Elderly Patients Undergoing Partial Hepatectomy.

Partial hepatectomy under general anesthesia is prone to hemodynamic alterations, and stress reactions are the main contributing factors to postoperative cognitive function in elderly partial hepatectomy patients. Postoperative cognitive dysfunction increases the incidence of postoperative complications and long-term morbidity and mortality in elderly patients. With the increasing trend of aging population and the gradual increase of elderly people undergoing surgical treatment, it is especially important to study the corresponding prevention and treatment measures. In this study, a total of 90 patients with primary liver cancer who received hepatectomy in our hospital from July 2020 to July 2021 were included as the research subject. The changes in hemorheology, stress-related indexes, cognitive function, postoperative pain, and gastrointestinal function were compared between the two groups The results showed that SGB combined with general anesthesia can effectively reduce hemodynamic fluctuations in elderly partial hepatectomy patients, alleviate surgical stress, promote postoperative recovery of cognitive function and gastrointestinal function with high safety, and is worthy of clinical promotion.

Genome-wide identification and characterization of olfactory receptor genes in common carp (Cyprinus carpio).

The environment contains a large extent of chemical information, which could be detected as olfactory sense. Olfactory in vertebrates plays important roles on many aspects during life time, including localizing prey or food, avoiding predators, mating behavior and social communication. Considering the essential role of olfactory receptors in the specific recognition of diverse stimuli, understanding the evolutionary dynamics of olfactory receptors in teleost means a lot, especially in the allotetraploid common carp, who has undergone the fourth whole-genome duplication event. Here, we identified the whole set of olfactory receptor genes in representative teleosts and found a significant contraction in common carp when compared with other teleosts. Odorant receptor genes (OR) occupy the most among four groups of olfactory receptors, including 33 functional genes and 16 pseudogenes. Furthermore, 6 trace amine-associated receptor (TAAR) genes (including 1 pseudogene), 7 odorant-related-A receptor genes, and 10 olfactory C family receptor genes (including 3 pseudogenes) were identified in common carp. Phylogenetic and motif analysis were performed to illustrate the phylogenetic relationship and structural conservation of teleost olfactory receptors. Selection pressure analysis suggested that olfactory receptor groups in common carp were all under relaxed purifying-selection. Additionally, gene expression divergences for olfactory receptor genes were investigated during embryonic development stages of common carp. We aim to determine the abundance of common carp olfactory receptor genes, explore the evolutionary fate and expression dynamics, and provide some genomic clues for the evolution of polyploid olfactory after whole-genome duplication and for future studies of teleost olfactory.

Chromosome-level genome of Poropuntius huangchuchieni provides a diploid progenitor-like reference genome for the allotetraploid Cyprinus carpio.

The diploid Poropuntius huangchuchieni in the cyprinid family, which is widely distributed in the Mekong and Red River basins, is one of the most closely related diploid progenitor-like species of allotetraploid common carp, which was generated by merging of two diploid genomes during evolution. Therefore, the P. huangchuchieni genome is essential for polyploid evolution studies in Cyprinidae. Here, we report a high-quality chromosome-level genome assembly of P. huangchuchieni by integrating Oxford Nanopore and Hi-C technologies. The assembled genome size was 1,021.38 Mb, 895.66 Mb of which was anchored onto 25 chromosomes with a N50 of 32.93 Mb. The genome contained 486.28 Mb repetitive elements and 24,099 protein-coding genes. Approximately 95.9% of the complete BUSCOs were detected, suggesting a high completeness of the genome. Evolutionary analysis revealed that P. huangchuchieni diverged from Cyprinus carpio at approximately 12 Mya. Genome comparison between P. huangchuchieni and the B subgenome of C. carpio provided insights into chromosomal rearrangements during the allotetraploid speciation. With the complete gene set, 17,474 orthologous genes were identified between P. huangchuchieni and C. carpio, providing a broad view of the gene component in the allotetraploid genome, which is critical for future genetic analyses. The high-quality genomic data set created for P. huangchuchieni provides a diploid progenitor-like reference for the evolution and adaptation of allotetraploid carps.

Transarterial chemoembolization combined with microwave ablation versus microwave ablation only for Barcelona clinic liver cancer Stage B hepatocellular carcinoma: A propensity score matching study.

PURPOSE: We aimed to compare the outcomes of microwave ablation (MWA) alone with those of transarterial chemoembolization combined with MWA (TACE-MWA) for Barcelona clinic liver cancer (BCLC) Stage B hepatocellular carcinoma (HCC) and to identify the prognostic factors associated with the two treatments. MATERIALS AND METHODS: This retrospective study was conducted in 150 BCLC Stage B HCC patients from April 2006 to November 2017. Of these, 88 patients were treated with MWA alone while 62 with TACE-MWA. Propensity score matching (PSM) was conducted to adjust for imbalances in clinical parameters. Procedure-related complications, local tumor progression (LTP), recurrence-free survival (RFS), and overall survival (OS) were analyzed. RESULTS: Before PSM, the maximal tumor diameters were 6.0 ± 1.0 cm and 6.7 ± 1.3 cm in the TACE-MWA and MWA groups, respectively, with a significant difference (P = 0.002); a significant difference was also detected in α-fetoprotein level (P = 0.013). After PSM, no difference was found in the two parameters (P = 0.067, 0.470). Before and after PSM, no difference was detected in the procedure-related complications (P = 0.803 vs. 1.000, P = 1.000 vs. 1.000), RFS (P = 0.786 vs. 0.689), and OS (P = 0.684 vs. 0.929). Tumor size and α-fetoprotein level were independent influencing factors for OS before and after PSM (P = 0.009, 0.023), while tumor size (D > 7) was an independent risk factor for poor OS (P = 0.011). Tumor number was an independent risk factor for RFS before and after PSM (P = 0.007 vs. P = 0.008). A significant difference was detected in LTP between the two groups with single tumor before and after PSM (P = 0.059 vs. P = 0.006). CONCLUSIONS: The MWA alone group had RFS and OS comparable to that of the TACE-MWA group. TACE-MWA was effective in controlling LTP in patients with a single tumor.

Ultrasound-guided percutaneous microwave ablation for 755 benign breast lesions: a prospective multicenter study.

OBJECTIVES: To evaluate the clinical efficacy of microwave ablation (MWA) of benign breast lesions (BBLs) and compare the learning curves of international radiologists (IRs) and surgeons. METHODS: In total, 440 patients with 755 clinicopathologically confirmed BBLs from 5 centers were prospectively enrolled from February 2014 to July 2018. Technical success, complications, volume reduction ratio (VRR), palpability, and cosmetic satisfaction after ablation were analyzed. In addition, the ablation time (AT) and energy (AE) with the number of procedures were analyzed for learning curve evaluation. RESULTS: The mean maximum diameter was 1.7 ± 0.6 cm. The complete ablation rate reached 100%, including 45.8% lesions adjacent to the skin, pectoralis, or areola. After a median follow-up of 13.7 months, the 12-month VRR of all lesions was 97.9%, and that for 1.0- to 2.0-cm and ≥ 2.0-cm lesions was 98.6% and 96.9%, respectively. A total of 55.9% of BBLs became nonpalpable (palpable in 85.7% of cases before MWA) by both the clinician and patient. The cosmetic and minimally invasive satisfaction rates were good or excellent in 98.4% and 94.5% of patients, respectively. The median AT/cm3 and AE/cm3 decreased as experience increased. The AE/cm3 of the IR with 5 years of experience was lower than that of the IR with 1 year of experience and the surgeons, while the AT/cm3 of surgeons was comparable with that of the IR with 5 years of experience at relatively mature phase. CONCLUSIONS: Ultrasound-guided percutaneous MWA is a valuable technique for the treatment of BBLs. TRIAL REGISTRATION: ClinicalTrials.gov (NCT02860104) KEY POINTS: • Ultrasound-guided percutaneous microwave ablation has the potential to become a valuable technique for the treatment of benign breast lesions. • A skilled interventional radiologist shows a rapid improvement in mastering the technique.

The allotetraploid origin and asymmetrical genome evolution of the common carp Cyprinus carpio.

Common carp (Cyprinus carpio) is an allotetraploid species derived from recent whole genome duplication and provides a model to study polyploid genome evolution in vertebrates. Here, we generate three chromosome-level reference genomes of C. carpio and compare to related diploid Cyprinid genomes. We identify a Barbinae lineage as potential diploid progenitor of C. carpio and then divide the allotetraploid genome into two subgenomes marked by a distinct genome similarity to the diploid progenitor. We estimate that the two diploid progenitors diverged around 23 Mya and merged around 12.4 Mya based on the divergence rates of homoeologous genes and transposable elements in two subgenomes. No extensive gene losses are observed in either subgenome. Instead, we find gene expression bias across surveyed tissues such that subgenome B is more dominant in homoeologous expression. CG methylation in promoter regions may play an important role in altering gene expression in allotetraploid C. carpio.

Effect of curcumin on vascular endothelial growth factor in hypoxic HepG2 cells via the insulin-like growth factor 1 receptor signaling pathway.

To investigate the anti-angiogenic effect and underlying molecular mechanisms of curcumin on HepG2 cells under hypoxic conditions, insulin-like growth factor 1 receptor (IGF-1R) knockout HepG2 cells were constructed using a clustered regularly interspaced short palindromic repeats/Cas9 genome-editing system. Hypoxic conditions were generated using cobalt chloride (CoCl2). An MTT assay was performed to measure the effects of curcumin on cell viability in hypoxia-induced IGF-1R knockout HepG2 cells, while western blot analysis was used to detect the expression of IGF-1R, phosphorylated (p)-protein kinase B (Akt), p-extracellular signal-regulated kinases (Erk)1/2, hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF). The results revealed that CoCl2 at low concentrations (50 and 100 µM) had no significant inhibitory effects on IGF-1R knockout HepG2 cells. However, with increasing concentrations of CoCl2 and treatment time, cell viability decreased and was significantly reduced at 150, 200 and 400 µM compared with the control group (P<0.05). The expression of HIF-1α and VEGF were significantly increased when the cells were treated with 150 or 200 µM CoCl2 compared with the control (P<0.05). With the increase of CoCl2 concentration or the treatment time, the expression of HIF-1α and VEGF were upregulated gradually. Additionally, curcumin significantly inhibited the expression of p-Akt, p-Erk1/2, HIF-1α and VEGF in hypoxia-induced IGF-1R knockout HepG2 cells. In conclusion, the findings of the present study suggest that curcumin may serve a pivotal role in tumor suppression via the inhibition of IGF-1R-mediated angiogenesis under hypoxic conditions.

Nickel(ii)-catalyzed tandem C(sp2)-H bond activation and annulation of arenes with gem-dibromoalkenes.

A nickel(ii)/silver(i)-catalyzed tandem C(sp2)-H activation and intramolecular annulation of arenes with dibromoalkenes has been successfully achieved, which offers an efficient approach to the 3-methyleneisoindolin-1-one scaffold. Attractive features of this system include its low cost, ease of operation, and its ability to access a wide range of isoindolinones.

Ultrasound guided percutaneous microwave ablation of benign breast lesions.

The benign breast lesions (BBLs) share a high incidence for women and therapy methods with minimal invasion and better cosmetic outcome are thirsted for. In this study, 122 patients with 198 biopsy-proved BBLs were enrolled. Ultrasound (US)-guided microwave ablation (MWA) was performed with local anesthesia from November, 2013 to April, 2016. The mean longest tumor size assessed was 1.6±0.7 cm (ranging 0.7-4.9 cm). MWA was successfully performed in all cases including 85 lesions adjacent to the skin, pectoralis and areola. The mean ablation time was 3.2mins (ranging 0.5-18.3 mins). 99.5% of BBLs showed complete ablation when assessed by magnetic resonance imaging and 100% of them by US. At the median 14-month follow-up, the BBLs were not palpable in 45.9 % of the cases (palpable in 90.2 % of the cases before MWA) and the mean volume reduction ratio was 78.4±33.5% for total lesions and 89.3±20.8%, 84.7±27.6% and 55.9±32.9% for ≤1.0 cm, 1.1-2.0cm and >2.0 cm lesions in 12-month follow-up, respectively. Cosmesis were reported as good or excellent in 100 % by physician and patients. No side effect was found. The MWA of the BBLs proved feasible and effective, while showing meaningful reduction in volume, palpability and cosmetic satisfying outcomes.

Genome-wide identification, phylogeny, and expression of fibroblast growth genes in common carp.

Fibroblast growth factors (FGFs) are a large family of polypeptide growth factors, which are found in organisms ranging from nematodes to humans. In vertebrates, a number of FGFs have been shown to play important roles in developing embryos and adult organisms. Among the vertebrate species, FGFs are highly conserved in both gene structure and amino-acid sequence. However, studies on teleost FGFs are mainly limited to model species, hence we investigated FGFs in the common carp genome. We identified 35 FGFs in the common carp genome. Phylogenetic analysis revealed that most of the FGFs are highly conserved, though recent gene duplication and gene losses do exist. By examining the copy number of FGFs in several vertebrate genomes, we found that eight FGFs in common carp have undergone gene duplications, including FGF6a, FGF6b, FGF7, FGF8b, FGF10a, FGF11b, FGF13a, and FGF18b. The expression patterns of all FGFs were examined in various tissues, including the blood, brain, gill, heart, intestine, muscle, skin, spleen and kidney, showing that most of the FGFs were ubiquitously expressed, indicating their critical role in common carp. To some extent, examination of gene families with detailed phylogenetic or orthology analysis verified the authenticity and accuracy of assembly and annotation of the recently published common carp whole genome sequences. Gene families are also considered as a unique source for evolutionary studies. Moreover, the whole set of common carp FGF gene family provides an important genomic resource for future biochemical, physiological, and phylogenetic studies on FGFs in teleosts.

[Studies on chemical constituents of Valeriana plants and their biological activities].

The recent progresses on chemical components and pharmacological activities of the genus Valerianawere summarized.Besides-essential oil, the chemical composition of Valerianais mainly focused on monoterpenoids,sesquiterpenoids,lignans, flavonoids, alkaloids, etc. Iridoids are the main chemical components ofmonoterpenoids. There are two types ofiridoidson the basis of the cyclopentane open or not. The Valerianahas been drawmuch attention for their significant sedation,spasmolysis,antidepression,antitumor, against adenosine A1 receptors and cytotoxicityactivity,and had certain function for cardiovascular disease treatment. Given to the fact of the lack of systematic review and summary of studies on the Valeriana, we summarized and analyze the study literatures on the pharmacological activity of Valerianain recent years, and providedsome basisfor further study.

Milkvetch root improves immune function in patients with acute exacerbation of COPD.

Milkvetch root as a medicine has been used for more over 2000 years in China, can strengthen immune function, protect liver, promote urination, resist aging and stress, reduce blood pressure and extensively resist bacterium. This study explored the effects of milkvetch root on the immune function of patients with a definitive diagnosis of acute exacerbation of chronic obstructive pulmonary disease (COPD). The patients were randomly assigned to either the experimental or control group. All patients received conventional clinical therapy; those in the experimental group were also administered milkvetch root. The serum levels of cytokines including tumor necrosis factor alpha (TNF-α), interleukin-8 (IL-8), IL-1ß, and IL-32 and immunocytes including T helper (Th), cytotoxic T (Tc), natural killer (NK), regulatory T (Treg) and B cells were measured 1 day before treatment and 7 and 14 days post-treatment. After bronchodilator inhalation, pulmonary function was evaluated at these same time points. The serum TNF-α, IL-8, IL-1ß, and IL-32 levels were significantly lower in the experimental group than in the control group 14 days post-treatment. The Th/Tc ratio and NK cell ratio was significantly higher but the Treg cell ratio was significantly lower in the experimental group than in the control group. The forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly higher in the experimental group than in the control group 14 days post-treatment. These results indicate that milkvetch root can improve the immune function of patients with acute exacerbation of COPD.

Hand-assisted laparoscopic versus open radical resection of gastric remnant cancer: a clinical comparison.

OBJECTIVE: To compare the perioperative clinical data between hand-assisted laparoscopic and open radical resection of gastric remnant cancer, and analyze the feasibility and advantages of hand-assisted laparoscopic resection of gastric remnant cancer. METHODS: The clinical data of 18 patients with gastric remnant cancer who underwent hand-assisted laparoscopic (n=9) or open surgery (n=9) were retrospectively analyzed to compare the perioperative outcomes and recovery between December 2007 and October 2014. RESULTS: All 18 patients underwent surgery without post-operative complications. In the hand-assisted laparoscopy group, none required conversion to open surgery, and no intraoperative auxiliary injury occurred. The incision length was (8.78±0.62) cm in the hand-assisted laparoscopy group, and (14.06±0.81) cm in the open surgery group (t=15.565, P=0.000). The duration of the operation averaged (221.11±19.48) min in the hand-assisted laparoscopy group, and (212.89±14.30) min in the open surgery group (t=-1.021, P=0.323). The intraoperative blood loss was (105.56±35.04) ml in the hand-assisted laparoscopy group, and (147.78±41.92) ml in the open surgery group (t=2.319, P=0.034). The number of lymph nodes scavenged was (16.22±2.99) and 16.67±3.28 in the hand-assisted laparoscopy group and in the open surgery group, respectively (t=-.300, P=0.768). The postoperative time to passage of gas by anus was (68.67±10.00) hr in the hand-assisted laparoscopy group, and (79.78±9.16) hr in the open surgery group (t=2.458, P=0.026). Mild postoperative hemorrhage occurred at the anastomotic site in one patient in each surgery group. These complications resolved after treatment. And no serious perioperative complications, such as anastomotic site fistula, massive hemorrhage in the peritoneal cavity or digestive tract or death occurred in either group. CONCLUSIONS: Hand-assisted laparoscopic radical resection of gastric remnant cancer is feasible, is of comparable efficacy to open surgery, and offers several advantages including small incisions, mild intraoperative hemorrhage, rapid postoperative recovery, and few postoperative complications.

Dynamic changes of matrix metalloproteinases in rat liver during the development of diethylnitrosamine-induced hepatocarcinoma.

OBJECTIVE: To study the dynamic changes of matrix metalloproteinases (MMPs) in the liver of rat during experimental hepatocarcinogenesis. METHODS: Immunohistochemistry, gelatin zymography, and reverse transcriptase-PCR were used for detection of latent and active forms and mRNA of MMPs in each phases of carcinogenic stages. RESULTS: MMPs expression was detected in normal and cirrhotic liver, which was most obvious in the cancer cells after the development of hepatic carcinoma. Normal tissue showed only low levels of MMPs expression, which kept increasing in the course of hepatocarcinogenesis, as was also the case with the corresponding mRNA. CONCLUSION: Transcription and expression of MMPs keep increasing throughout the the process of hepatocarcinogenesis.