Electroacupuncture promotes recovery from disuse muscular atrophy through the miR-133a/SIRT1 pathway / 中华物理医学与康复杂志

Objective:To explore the role of microRNA-133a (miR-133a) and silent mating information regulation 2 homolog 1 (SIRT1) in the effects of electroacupuncture on persons with disuse muscular atrophy.Methods:Thirty C57BL/6 mice were randomly divided into a control group, an experimental control group and an experimental group, each of 10. Disuse muscular atrophy was induced in the mice of the experimental and experimental control groups using tail suspension. The mice in the electroacupuncture group were given 15 minutes of electroacupuncture over the Yanglingquan and Zusanli points every day for 14 days. Wet weight ratio and the cross-sectional area of the gastrocnemius and soleus were tracked, and the structure of the mitochondria in the skeletal muscles was observed using a transmission electron microscope. The protein expressions of SIRT1, peroxisome proliferator-activated receptor γ coactivator-1a (PGC-1a), nicotinamide phosphoribosyl transferase (NAMPT), adenosine 5-monophosphate-activated protein kinase-a (AMPK-a) and phospho-AMPK-a (P-AMPK-a) were detected using western blotting. The expressions of the muscle atrophy F-box (Atrogin-1), muscle ring finger1 (MuRF1), miR-133a, SIRT1, paired box gene 7 (Pax7), myogenic determination (MyoD) and myogenin (MyoG) genes were detected through polymerase chain reactions. The concentration of Niacylamide adenine dinucleotide (NAD+ ) and the ratio of NAD+ to reduced nicotinamide adenine dinucleotide were also measured.Results:Compared with the experimental control group, the average wet weight increased by 21% and the cross-sectional area of the soleus increased by 30% in the experimental group. The average wet weight of the gastrocnemius increased by 5% and the area by 17%. The average expressions of Atrogin-1, MuRF1, SIRT1, PGC-1a and NAMPT, the concentration of NAD+ , as well as the average value of P-AMPK-a/AMPK-a and NAD+ /NADH were significantly lower in the experimental group than in the experimental control group, while the average expression of miR-133a in the experimental group was significantly (163%) higher. The average expressions of Pax7 and MyoD were significantly up-regulated in the experimental control group compared with the other two groups, while MyoG was highly expressed in the experimental group compared with the other 2 groups.Conclusions:The SIRT1 pathway is one of the reflexive protective mechanisms that mediate in the natural recovery of skeletal muscles. Electroacupuncture enhances myoblast differentiation, improves energy metabolism in the mitochondria, and thus promotes recovery from disuse muscular atrophy.

Effects of Incremental Load Training on Phosphorylation of AMPK in Skeletal Muscle Satellite Cells of Aged Mice / 医用生物力学

Objective To investigate the effect of incremental load training on AMP-activated protein kinase (AMPK) phosphorylation in skeletal muscle satellite cells of aged mice. Methods Experimental mice were divided into 3 groups: young control group (YC group, n=12), old control group (OC group, n=12) and old training group (OT group, n=12). The mice in OT group received incremental load training, and CD45-/CD31-/Sca1-/VCAM (CD106) + cells were isolated by flow cytometry sorting. Desmin, Myod myogenic staining and myogenic differentiation culture were used for identification of muscle satellite cells, and the p-AMPK level of muscle satellite cells was detected by immunohistochemistry combined with Western blotting method. Results The expression levels of AMPK and p-AMPK in skeletal muscle satellite cells in YC group were significantly higher than those in OC group (P0.05), while p-AMPK expression level in OT group was significant higher than that in OC group (P<0.05). Conclusions Incremental load training can promote AMPK phosphorylation of skeletal muscle satellite cells in aged mice, and improve energy metabolism of skeletal muscle in aged mice.

The mechanism through which low-intensity pulsed ultrasound promotes the differentiation of C2C12 myoblasts / 中华物理医学与康复杂志

Objective:To observe the effect of low-intensity pulsed ultrasound (LIPUS) at different intensities on the expression of adiponectin and its receptors in C2C12 myoblasts, and to explore the potential mechanism by which LIPUS promotes the differentiation of C2C12 myoblasts.Methods:C2C12 myoblasts cultured in vitro were randomly divided into a control group and U 0.1, U 0.3 and U 0.5 groups. The control group received sham LIPUS exposure, while the U 0.1, U 0.3 and U 0.5 groups were exposed to LIPUS at intensities of 0.1W/cm 2, 0.3W/cm 2 or 0.5W/cm 2 respectively, and 1MHz for 5 min daily for 5 days. Cell viability was measured using CCK-8 assays. Fluorescence quantitative reverse transcription-polymerase chain reactions were used to detect the mRNA expression of adiponectin, adiponectin receptor 1 (adipoR1) and T-cadherin in the cells. Western blotting was employed to assess the protein expression of adiponectin, adipoR1, T-cadherin, adenosine monophosphate activated protein kinase (AMPK), activated phosphorylated adenylate-activated protein kinase (P-AMPK), embryonic myosin heavy chain (eMHC) and myogenin (MYOG). The differentiation ability of the 4 groups was measured using cell immunofluorescence chemistry. Results:After the intervention the cell viability in the U 0.1, U 0.3 and U 0.5 groups was significantly higher than in the control group. Compared with the control group, the average mRNA expression of adiponectin and the receptors of adipoR1 and T-cadherin were up-regulated significantly in the U 0.3 and U 0.5 groups. The average adiponectin, adipoR1 and T-cadherin protein expressions, and the AMPK phosphorylation level in the U 0.3 and U 0.5 groups had increased significantly compared with the control group, but all were significantly lower than in the U 0.3 group. The average protein expression of eMHC and MYOG, and the C2C12 myoblast fusion indices of the U 0.3 and U 0.5 groups were significantly higher the control group′s averages. Conclusions:LIPUS can promote the differentiation of C2C12 myoblasts. It is most effective at 0.3W/cm 2, administered for 5min/d at 1MHz with a 20% duty cycle. Its regulatory mechanism may be related to up-regulation of the expression of adiponectin, the adipoR1 and T-cadherin receptors, and the activation of AMPK phosphorylation in C2C12 myoblasts.

Upregulation of OSBPL3 by HIF1A promotes colorectal cancer progression through activation of RAS signaling pathway.

Oxysterol-binding protein like protein 3 (OSBPL3) has been shown involving in the development of several human cancers. However, the relationship between OSBPL3 and colorectal cancer (CRC), particularly the role of OSBPL3 in the proliferation, invasion and metastasis of CRC remains unclear. In this study, we investigated the role of OSBPL3 in CRC and found that its expression was significantly higher in CRC tissues than that in normal tissues. In addition, high expression of OSBPL3 was closely related to poor differentiation, advanced TNM stage and poor prognosis of CRC. Further experiments showed that over-expression of OSBPL3 promoted the proliferation, invasion and metastasis of CRC in vitro and in vivo models. Moreover, we revealed that OSBPL3 promoted CRC progression through activation of RAS signaling pathway. Furthermore, we demonstrated that hypoxia induced factor 1 (HIF-1A) can regulate the expression of OSBPL3 via binding to the hypoxia response element (HRE) in the promoter of OSBPL3. In summary, Upregulation of OSBPL3 by HIF1A promotes colorectal cancer progression through activation of RAS signaling pathway. This novel mechanism provides a comprehensive understanding of both OSBPL3 and the RAS signaling pathway in the progression of CRC and indicates that the HIF1A-OSBPL3-RAS axis is a potential target for early therapeutic intervention in CRC progression.

The effects of endurance exercise on myocardial T-cadherin in aged mice / 中华物理医学与康复杂志

Objective:To observe the effect of endurance exercise on the expression of T-cadherin, its ligand adiponectin and related downstream molecules in the myocardia of aged mice, and to explore the role of T-cadherin in the observed effects.Methods:Thirty 17-month-old C57 male mice were divided randomly into a control group and an exercise group, each of 15. Another 15 2-month-old C57 male mice constituted the young control group. The exercise group performed endurance exercises for 12 weeks, while no exercise was taken in the two control groups. All of the mice were then sacrificed within 24 hours after the exercise group′s last exercise session and the expression of T-cadherin and its ligand adiponectin was measured along with the level of phosphorylation of AMPK, apoptosis and the expression of the autophagy-related signaling proteins bax, bcl-2, Beclin-1 and p-mTOR using western blotting. Immunohistochemistry was used to observe the location and positive expression of T-cadherin, adiponectin and p-mTOR cells in the myocardium.Results:The T-cadherin was localized in the membrane and vascular endothelia of cardiomyocytes. Adiponectin was localized in the membrane, cytoplasm and vascular endothelium, and p-mTOR in the cytoplasm. Compared with the young control group, the average T-cadherin, adiponectin, AMPK activation level, apoptotic protein Bcl-2 and autophagy-promoting protein Beclin-1 expression of the aged control group was significantly lower, while the average level of the apoptotic protein bax, Caspase-9 expression, and the phosphorylation of autophagy-related protein mTOR were significantly elevated. Compared with the older control group, the average T-cadherin and ligand adiponectin, AMPK activation level, and Bcl-2 and Beclin-1 protein expression in the exercise group were significantly greater, while the average expression of bax and Caspase-9 protein was significantly lower, as was mTOR phosphorylation.Conclusions:Endurance exercise can increase the expression of T-cadherin in the myocardium, promote the binding of adiponectin to myocardial tissue, and stimulate the activation of AMPK, playing a role in protecting the myocardium, at least in elderly mice.

Embryo-fetal developmental toxicity and toxicokinetics of carenoprazan hydrochloride in rabbits / 中国药理学与毒理学杂志

OBJECTIVE To investigate the embryo-fetal developmental toxicity (EFDT) of careno?prazan hydrochloride (KFP-H008) in rabbits. METHODS Pregnant rabbits were given by gavage KFP-H008 at 5, 15 and 50 mg·kg-1 during the organogenetic period (gestation days 6-18, GD 6-18). Rabbits in positive control group were treated with cyclophosphamide (CP) 10 mg·kg-1 by iv. Maternal body mass and food consumption during gestation were recorded. Pregnant dams were euthanized on GD 29. The numbers of live/dead fetuses, resorptions, implantations, corpora lutea, and gravid uterus mass, placenta mass, fetal gender ratios, body mass, and skeletal development were evaluated. Moreover, the toxicokinetic parameters including AUC and C0-t, and tissue distributions were determined. RESULTS From GD 13, the maternal body mass and the food consumption in KFP-H00815 and 50 mg · kg-1 groups were lower than in the normal control group (P<0.05). Also, the reduced fetal crown rump length and mass, skeletal malformations/variations were observed in KFP-H00815 and 50 mg · kg-1 groups (P<0.05). KFP-H008 was rapidly eliminated, and became undetectable in the maternal plasma after a single administration. Following multiple KFP-H00850 mg · kg-1 treatment, both KFP-H008 and its metabolites were detectable in various tissues of the maternal and fetus, which might be the evidence for carenoprazan-induced developmental toxicity. In KFP-H00815 mg · kg-1 group, KFP-H008 and its metabolites were undetectable in most of maternal and fetal tissues. CONCLUSION The no observed adverse effect level (NOAEL) of KFP-H008 for maternal and fetal rabbits is about 5 mg·kg-1.

Treatment of intrahepatic cholangiocarcinoma / 国际外科学杂志

Intrahepatic cholangiocarcinoma has low resectability rate, high recurrence and short survival. It is very important to formulate and optimize the strategy of surgical treatment. The only potentially effective treatment for intrahepatic cholangiocarcinoma is surgical resection. Liver transplantation also has some application prospects. Intrahepatic cholangiocarcinoma can be divided into four types: mass forming type, intraductal growth type, periductal infiltration type, mass forming + periductal infiltration(mixed)type. Clinically, the treatment strategy is mainly determined according to the general classification. The application of methods such as preoperative portal vein embolism, neoadjuvant therapy and lymph node dissection make it possible for more patients to undergo surgical resection and improve the surgical effect. Adjuvant treatment including chemotherapy and radiotherapy can significantly improve the prognosis of the patients. The rapid development of molecular targeted therapy and immunotherapy is gradually changing the clinical treatment of intrahepatic cholangiocarcinoma.

Candidate genes involved in spiroacetal biosynthesis in the oriental fruit fly, Bactrocera dorsalis.

Spiroacetals are widespread in nature as components of volatile semiochemical secretions from many insect species. The general pathway for spiroacetal biosynthesis in Bactrocera sp. is preliminarily established, but many genes involved in this pathway remain to be characterized. By analyzing transcriptomes of the rectal glands (RGs) from immature and mature females of the oriental fruit fly, Bactrocera dorsalis, a set of genes encoding two acetyl-CoA carboxylases (ACCs), two fatty acid synthases (FASs), eight desaturases (DESs), twelve fatty acyl-CoA reductases (FARs), seventy-two cytochrome P450 enzymes (CYPs), and twenty-three odorant binding proteins (OBPs) were identified. We investigated the expression of candidate genes in immature and mature stages based on the RNA-seq data and Real-time quantitative PCR. Expression profiling revealed that some of these genes were primarily expressed in female rectal glands among different tissues, and were up-regulated in mature females. Semi-quantitative RT-PCR assays were also adapted to examine tissue-specific expression of selected candidate genes. Additionally, their putative functions in spiroacetal synthesis and transportation are proposed. Our study provided large-scale sequence information for further functional studies on spiroacetal biosynthetic pathways.

Candidate genes coding for odorant binding proteins and chemosensory proteins identified from dissected antennae and mouthparts of the southern green stink bug Nezara viridula.

The southern green stink bug Nezara viridula (Hemiptera: Pentatomidae) is a highly polyphagous pest that can significantly impact many major crops worldwide. Insect odorant binding proteins (OBPs) and chemosensory proteins (CSPs) transport chemicals and play critical roles in chemoreception. Studies on N. viridula OBPs and CSPs should increase our overall understandings on chemosensory systems and chemical ecology of stink bugs, which may lead to improved pest control. In this study, we identified candidate genes encoding putative OBPs and CSPs in N. viridula by generating transcriptomes from dissected antennae and mouthparts. In total, the 42 unigenes were identified coding for OBPs (34 Classic OBPs and eight Plus-C OBPs) and 13 unigenes coding for CSPs. Expression profiles of OBP- and CSP -encoding genes were compared between antennae and mouthparts based on FKPM values. Candidates for antenna-predominant OBPs and CSPs were selected for real-time quantitative PCR analyses. Analyses of tissue expression profiles revealed that 17 OBP-encoding genes, and four CSP genes were primarily expressed in antennae, suggesting their putative roles in perception of volatiles. The sex-biased expression patterns of these antenna-predominant genes suggested that they may have important functions in reproduction of the insect. This is a systematic analysis on OBPs and CSPs in a stink bug, providing a comprehensive resource for future functional studies not only for N. viridula, but also for other stink bugs as well.

The effect of aerobic endurance exercise on renal fibrosis and the expression of TGF-beta 1 and E-cadherin in aged mice / 中华物理医学与康复杂志

Objective@#To explore the effect of aerobic endurance exercise on renal fibrosis in elderly mice and its possible mechanism.@*Methods@#Thirty-six healthy, male C57 mice were sorted into a young control group (2 months old), a senile control group (19 months old) and a senile exercise group (also 19 months old). The senile exercise group underwent aerobic endurance exercise for 7 weeks. Then all of the mice were sacrificed and any changes of in their renal tissues were recorded, especially the expression of fibrosis indicators using HE staining, Sirius red staining, RT-PCRs and immunohistochemical methods.@*Results@#Collagen fibers deposited in the kidney tissue of the senile groups were significantly more numerous than in the young control group, but their number had decreased significantly after the exercise.@*Conclusion@#Aerobic endurance exercise can inhibit collagen deposition and delay the formation of renal fibrosis, at least in rats. This may be related to its inhibition of TGF-β1 and α-SMA expression and up-regulation of E-cadherin expression.

Summary of the 2019 Academic Annual Meeting of the Chinese Burn Association / 中华烧伤杂志

The 2019 Academic Annual Meeting of the Chinese Burn Association, sponsored by the Chinese Medical Association and the Chinese Burn Association, was successfully held in Zhuhai, Guangdong province, from November 6th to 9th, 2019. The theme of this conference was " One China, One Standard--Data Standardization and Construction of National Burn Data Platform" . A total of 2 305 submissions and 1 749 e-posters were received, and 1 097 registered representatives, nearly 2 000 representatives from 9 countries and regions attended the meeting. Focusing on the theme of this conference, a variety of novel forms were adopted such as teaching contest of young surgeons, multi-disciplinary discussion, workshop, and surgery live broadcast on hot issues in key areas of burns. Besides, with the focus on humanistic care and innovation, a multi-disciplinary discussion was warmly conducted. The 2020 academic annual conference is scheduled to be held in Nanchang, China.

Embryo-fetal development toxicity of carenoprazan hydrochloride / 中国药科大学学报

@#Carenoprazan has the similar structure and mechanism with the potassium-competitive blocker vonoprazan. Howerver, its safety during the pregnancy remains uncertain. To study the embryo-fetal development toxicity and toxicokinetics of carenoprazan hydrochloride via oral administration, time-mated Sprague-Dawley rats were divided into 5 groups, treated with normal saline, cyclophosphamide for injection(3. 8 mg/kg), and carenoprazan hydrochloride(20, 60, 200 mg/kg), respectively. Administrated orally from gestation day(GD)6 - 15. At the termination(GD 20), pregnant dams were sacrificed, and concentrations of carenoprazan hydrochloride as well as its metabolite in plasma and issues of both maternal and fetus were examined. As a result, the body weight gain of maternal in both high(200 mg/kg)and medium(60 mg/kg)dose as well as the food consumption of high-dose were decreased during GD 10-16. At the high dose group, decrease of crown rump length of fetuses were significant. Also, skeletal malformation/variations of fetus increased obviously at both high- and medium- dosage. The toxcicokinetics of carenoprazan hydrochloride are linear after single treatment between 20-200 mg/kg. The placental barrier was penetrated by carenoprazan hydrochloride and metabolite, and the distribution of metabolite in organs were similar in both maternal and fetus, with the highest concentration in livers. Therefore might resulted in the development toxicity. The No Observed Adverse Effect Level(NOAEL)of carenoprazan hydrochloride for both maternal and fetal was 20 mg/kg.

Summary of the 2018 Academic Annual Meeting of the Chinese Burn Association / 中华烧伤杂志

The 2018 Academic Annual Meeting of the Chinese Burn Association, sponsored by the Chinese Medical Association and the Chinese Burn Association, was successfully held in Fuzhou, Fujian Province, from October 24th to 27th. The theme of this conference is " One China, One Standard". A total of 1, 798 submissions were received, and 1, 060 registered representatives, more than 2, 000 representatives from 9 countries and regions attended the meeting. Focusing on the theme of " One China, One Standard" , the conference adopted a variety of innovative forms such as academic debate, live surgery, BBS on both sides of the straits, award selection, and so on to provide participants with multiple ways for exchange on the professional hot issues in the key areas of burns. The atmosphere of the conference was warm. The 2019 annual academic conference is scheduled to be held in Zhuhai, China.

Antennal transcriptome and expression analyses of olfactory genes in the sweetpotato weevil Cylas formicarius.

The sweetpotato weevil, Cylas formicarius (Fabricius), is a serious pest of sweetpotato. Olfaction-based approaches, such as use of synthetic sex pheromones to monitor populations and the bait-and-kill method to eliminate males, have been applied successfully for population management of C. formicarius. However, the molecular basis of olfaction in C. formicarius remains unknown. In this study, we produced antennal transcriptomes from males and females of C. formicarius using high-throughput sequencing to identify gene families associated with odorant detection. A total of 54 odorant receptors (ORs), 11 gustatory receptors (GRs), 15 ionotropic receptors (IRs), 3 sensory neuron membrane proteins (SNMPs), 33 odorant binding proteins (OBPs), and 12 chemosensory proteins (CSPs) were identified. Tissue-specific expression patterns revealed that all 54 ORs and 11 antennal IRs, one SNMP, and three OBPs were primarily expressed in antennae, suggesting their putative roles in olfaction. Sex-specific expression patterns of these antenna-predominant genes suggest that they have potential functions in sexual behaviors. This study provides a framework for understanding olfaction in coleopterans as well as future strategies for controlling the sweetpotato weevil pest.

Antennal and abdominal transcriptomes reveal chemosensory gene families in the coconut hispine beetle, Brontispa longissima.

Antennal and abdominal transcriptomes of males and females of the coconut hispine beetle Brontispa longissima were sequenced to identify and compare the expression patterns of genes involved in odorant reception and detection. Representative proteins from the chemosensory gene families likely essential for insect olfaction were identified. These include 48 odorant receptors (ORs), 19 ionotropic receptors (IRs), 4 sensory neuron membrane proteins (SNMPs), 34 odorant binding proteins (OBPs) and 16 chemosensory proteins (CSPs). Phylogenetic analysis revealed the evolutionary relationship of these proteins with homologs from Coleopterans or other insects, and led to the identification of putative aggregation pheromone receptors in B. longissima. Comparative expression analysis performed by calculating FPKM values were also validated using quantitative real time-PCR (qPCR). The results revealed that all ORs and antennal IRs, two IR co-receptors (BlonIR8a and BlonIR25a) and one SNMP (BlonSNMP1a) were predominantly expressed in antennae when compared to abdomens, and approximately half of the OBPs (19) and CSPs (7) were enriched in antennae. These findings for the first time reveal the identification of key molecular components in B. longissima olfaction and provide a valuable resource for future functional analyses of olfaction, and identification of potential targets to control this quarantine pest.

Transcriptome sequencing of Tessaratoma papillosa antennae to identify and analyze expression patterns of putative olfaction genes.

Studies on insect olfaction have increased our understanding of insect's chemosensory system and chemical ecology, and have improved pest control strategies based on insect behavior. In this study, we assembled the antennal transcriptomes of the lychee giant stink bug, Tessaratoma papillosa, by using next generation sequencing to identify the major olfaction gene families in this species. In total, 59 odorant receptors, 14 ionotropic receptors (8 antennal IRs), and 33 odorant binding proteins (28 classic OBPs and 5 plus-C OBPs) were identified from the male and female antennal transcriptomes. Analyses of tissue expression profiles revealed that all 59 OR transcripts, 2 of the 8 antennal IRs, and 6 of the 33 OBPs were primarily expressed in the antennae, suggesting their putative role in olfaction. The sex-biased expression patterns of these antenna-predominant genes suggested that they may have important functions in the reproductive behavior of these insects. This is the first report that provides a comprehensive resource to future studies on olfaction in the lychee giant stink bug.

The effect of calorie restriction on the proliferation of satellite cells in elderly rats' skeletal muscles / 中华物理医学与康复杂志

Objective To explore any effect of calorie restriction on the proliferation of satellite cells in the skeletal muscles of elderly rats.Methods Twelve male C57BL rats aged 12 or 13 months were randomly divided in to an experimental group and a control group,each of 6.The control group was fed 75.09 kJ/d as normal,while the experimental group was provided with 45.05 kJ/d (60％ of normal).The intervention lasted for 15 weeks and each rat's weight was measured every week.After the intervention,limb muscle satellite cells were sorted by fluorescenceactivated cell sorting after digestion,and the cell cycle was analyzed.Western blotting was used to assess the expression of cyclin A,D1 and E.Results There was no significant difference in the average weight of the two groups before the experiment.After the 15 weeks the average weight of the experimental group had decreased significantly (to 19.5±0.4 g),and it was significantly lighter than that of the control group (31.9±0.5 g).The average percentage of the satellite cells in the G0/G1 phase had decreased significantly in the experimental group,but the percentage in the S phase had increased significantly.The expression of cyclin A and E was significantly greater in the experimental group compared with the control group,but the expression of cyclin D1 had decreased significantly.Conclusion Caloric restriction can delay the proliferation of satellite cells in the skeletal muscles of elderly mice.

Correlation Study Between Autoantibodies to Myocardium β1-adrenergic Receptor, M2-muscarinic Receptor and Hypertrophic Cardiomyopathy / 中国循环杂志

Objective: To study the relationship between autoantibodies to myocardium β1-adrenergic receptor (β1-receptor), M2-muscarinic receptor (M2-receptor) and hypertrophic cardiomyopathy (HCM) in relevant patients. Methods: Our research included in 2 groups: HCM group, n=81 patients and Control group, n=50 normal subjects. Synthetic peptides corresponding to amino acid sequence of second extracellular loops of β1-receptor and M2-receptor were used as antigens, ELISA was conducted to detect serum autoantibodies to β1-receptor and M2-receptor in both groups. Results: ① Compared with Control group, HCM group had the higher positive rates of autoantibodies as for β1-receptor was 58.02% (47/81) vs 12.00% (6/50) and for M2-receptor was 44.44 % (36/81) vs 10.00% (5/50), both P<0.05. ② Compared with Control group, HCM group had the higher mean geometric titers of autoantibodies as for β1-receptor was 1:85 vs 1:36 and for M2-receptor was 1:80 vs 1:35, both P<0.05. ③ In HCM group, 43.48 % patients with positive autoantibody to β1-receptor meanwhile have autoantibody to M2-receptor. Conclusion: HCM patients having both autoantibodies to β1-receptor and M2-receptor might be related to cardiac structure change and diastolic function decreasing. Both positive antibodies implies that immunological mechanism has been involved in pathophysiological process in HCM patients.

Construction of nuclear factor of activated T-cells 5 mRNA 3'-untranslated region reporter vector and targeting verification between NFAT5 and miR-155 / 重庆医学

Objective To construct a Luciferace reporter vector containing the 3'untranslated region (3'UTR) of NFAT5 and measure the correlation between NFAT5 and miR-155.Methods The miR-155 targeting NFAT5 3'UTR was predicted by Target Scan,Mir Base and Pic Tar.NFAT5 and mutant NFAT5 sequence(NFAT5-mu) were then designed and synthesized,and they were cloned into pMIR-REPORTTM Luciferace reporter vector.Human embryonic kidney-293AD (HEK-293AD) cells of the 4th passage were divided into 4 groups according to the random number table.cells in plasimd +miR-155 mimics groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 mutated groups were transfected with pMIR-NFAT5-mu recombinant plasimid,pRL-Tk plasmid and miR-155 mimics;cells in plasimd + miR-155 control groups were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 Negative control;cells in plasimd +miR-155 inhibitor were transfected with pMIR-NFAT5 recombinant plasimid,pRL-Tk plasmid and miR-155 inhibitor;and were respectively transfected into together by liposome.After culture for 24 h,the luciferase activity was detected by dual luciferase reporter assay system.Results TargetScan,Miranda and PicTar shared the results that NFAT5 has the complementary binding sites with 3'UTR of miR-155.And luciferase reporter vectorwas constructed.Therefore the result of sequencing and double digesting of recombined plasmid were completely correct.Dual-luciferase reporter assay showed that miR-155 possesses a target effect on 3'UTR of NFAT5.Compared to the pMIR-NFAT5 + miR-control group,the luciferase activity of the pMIR-NFAT5 + miR-1 5 5 mimics group was decreased,with statistically significant difference(P＜0.01),while there was no significant difference at other time points(P＞0.05).Conclusion The pMIR-NFAT5 recombinant plasmid and pMIR-NFAT5 recombinant mutated plasmid were confirmed with successful construction.and it was found that miR-155 can target NFAT5 mRNA 3'-UTR.The results provide the experiment data for further disclosing the mechanism of inhalation injury on the level of gene expression.