Enhancement of tropane alkaloids biosynthesis in Atropa belladonna hariy root by overexpression of HnCYP82M3 and DsTRI genes / 药学学报

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.

Mei mini maze procedure for atrial fibrillation patients with atrial functional mitral regurgitation / 中华胸心血管外科杂志

Objective:To study the influence of Mei mini maze procedure for atrial functional mitral regurgitation.Methods:The data of 33 patients with atrial fibrillation and atrial functional mitral regurgitation from January 2017 to June 2020 were retrospectively analyzed. All patients received Mei mini maze procedure for atrial fibrillation. The procedure is carried out thoracoscopically through the left thoracic approach. The ablation of atrial fibrillation includes bilateral circumferential pulmonary vein ablation, isolation of the left atrium posterior wall, left atrial appendage resection, ablation of Marshall's ligament and autonomic ganglion, etc. Follow-up was conducted by outpatient follow-up and telephone. Postoperative heart rhythm was recorded by the patient's symptoms, electrocardiogram, 24 h holter and other examinations. Postoperative mitral valve lesions were obtained by echocardiography.Results:33 patients successfully completed the operation. There was no conversion to thoracotomy and no perioperative death. Thirty patients(90.9%) maintained sinus rhythm at discharge. Before discharge, 16 patients had no mitral regurgitation in echocardiography, 8 patients had mild mitral regurgitation, and 9 patients had moderate mitral regurgitation. Follow-up was 1-4 years after discharge, with a mean of(2.6±1.1) years. Sinus rhythm was maintained in 23 patients(69.7%). 17 patients had no mitral regurgitation, 9 had mild mitral regurgitation, 6 had moderate, and 1 had severe mitral regurgitation. The degree of regurgitation in 25 patients was reduced compared with pre-operation, 5 patients remained unchanged, and 3 patients mitral regurgitation aggravated. Unreduced atrial functional mitral regurgitation was associated with recurrence of atrial fibrillation by Cox multivariate analysis.Conclusion:This study found a close relationship between atrial fibrillation rhythm and atrial functional mitral regurgitation. Most moderate atrial functional mitral regurgitation can be alleviated by effective treatment for atrial fibrillation. It is not recommended that patients with severe atrial functional mitral regurgitation only receive treatment for atrial fibrillation.

Progress in research of etiology of childhood obesity based on interaction between genes and environment / 中华流行病学杂志

Childhood obesity is a global public health problem, which can not only endangers children's health, but also might be an important cause of chronic diseases in adulthood. In recent years, with the in-depth development of precision medicine research, more and more research evidences have shown that there are interactions between environmental factors, such as early intrauterine environment, children's diet, physical activity and children's gene factor on the incidence of childhood obesity, which can result in or inhibit the incidence and development of childhood obesity. This paper summarizes the progress in research in this field to reveal the effects and potential mechanisms of genetic factors and environmental factors on the incidence of childhood obesity in order to provide reference for the precise prevention and control of childhood obesity under different genetic backgrounds.

Visual Detection of Plant Viral RNA Based on Gold Nanoparticles and Hybrid Chain Reaction / 中国生物化学与分子生物学报

The harm of plant virus disease is serious, which significantly restricts the sustainable development of agriculture and can cause huge economic losses. Monitoring plant health and early detection of viral pathogens are essential to reduce the spread of disease. Therefore, in order to realize the early detection of plant viral diseases in the field, a sensitive, specific and efficient colorimetric visual technique for plant viral RNA was designed by combining the colorimetric method based on gold nanoparticles (AuNPs) and hybrid chain reaction (HCR). In this study, tobacco mosaic virus (TMV) was used as a model to design two hairpin structures H1/ H2 with single-stranded tails based on TMV-specific conserved fragments. TMV could open the hairpin structure to alternately form long double-straight DNA. The binding difference between AuNPs and two nucleic acid states before and after HCR reaction resulted in colorimetric signal generation, thus realizing visual detection of TMV. After optimizing the concentration of Tris-HAc, the concentration of hairpin structure and the reaction time of HCR, the best detection conditions were obtained. The sensitivity and specificity of the technique were analyzed and real samples were tested under optimal conditions. The results showed that the absorbance ratio of AuNPs (A

Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson' s disease model of mice induced by paraquat / 解剖学报

Objective To investigate the protective effect and mechanism of liraglutide on the paraquat (PQ)⁃ induced Parkinson's disease (PD) mouse model. Methods Totally 24 Kunming mice were randomly divided into control group, PQ group and PQ +liraglutide group, 8 mice in each group. PD model was established by intraperitoneal injection of PQ (10 mg/kg) for 5 consecutive days, and liraglutide (50 nmol/kg) was injected intraperitoneally for 7 consecutive days. The free⁃standing and locomotor activity of mice were measured by behavioral method. Immunofluorescence was used to observe the number of tyrosine hydroxylase (TH) and ionized calcium binding adaptor molecule 1 (Iba1) immunoreactive cells. Western blotting was used to detect the expression of protein TH, glial fibrillary acidic protein (GFAP), mitofusin⁃2 (Mfn2) and dynamin⁃related protein 1 (Drp1). Results The numbers of free⁃standing and locomotor activity numbers decreased significantly (P<0.01, P < 0.05) in PQ group compared with the control group, and the number of TH immunoreactive cells and TH protein expression in substantia nigra decreased significantly (P<0.01, P<0.01) compared with the control group, while the number of Iba1 immunoreactive cells and GFAP protein expression increased significantly (P<0.01, P<0.01) compared with the control group; the expression of Drp1 protein in PQ group was significantly higher than that in control group (P<0.05), while the Mfn2 protein expression decreased significantly (P<0.05) compared with the control group. After treatment with liraglutide, the number of TH positive cells in PQ + liraglutide group was significantly lower than that in control group (P<0.05); the numbers of free⁃standing and locomotor activity increased significantly (P<0.05, P<0.05) in PQ + liraglutide group compared with the PQ group, and the number of TH positive cells and expression of TH protein in PQ + liraglutide group were significantly higher than that in PQ group (P<0.01, P< 0.01); while the number of Iba1 positive cells and GFAP protein expression decreased significantly (P<0.01, P<0.05) compared with the PQ group; the Drp1 protein expression decreased significantly (P<0.01) compared with the PQ group, while the expression of Mfn2 protein in PQ + liraglutide group was significantly higher than that in PQ group (P<0.01). Conclusion Liraglutide has neuroprotective effect by reducing neuroinflammation in substantia nigra, regulating mitochondrial fusion and fission.

Effect of the rolB gene on phenotypic development and tropane alkaloids biosynthesis in Atropa belladonna / 药学学报

The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

Verbal and nonverbal cognitive function of aphasia after stroke based on Web of Science database: a visualized analysis / 中国康复理论与实践

ObjectiveTo visualize the research trends of verbal and nonverbal cognitive functions in patients with post-stroke aphasia (PSA). MethodsPapers about verbal and nonverbal cognitive functions after stroke were extracted from Web of Science Core Collection from establishment to August, 2022, and a bibliometric analysis was conducted on the distribution of articles published by year, author, journal, institution, and country. Author cooperation, author co-citation, journal co-citation, country/institution cooperation, dynamic evolution, keyword hotspots, and trends were visually analyzed by using VOSviewer software. ResultsA total of 1 630 publications were identified for final analysis. The first article related to the topic was retrieved in 2004. After that, the number of published papers showed an increasing trend year by year, with some fluctuations after 2019. Matthew A. Lambon Ralph and Elizabeth Jefferies were the authors who produced the most, were cited the most, and were the most influential. Neuropsychologia was a highly productive and well-cited publication. The University of Manchester in the United Kingdom and Johns Hopkins University in the United States were the nations/institutions with high yield and high centrality. Although China had a high yield and was in the top five, the centrality was insufficient. The dynamic emergence of keywords revealed that the research had gone through the following stages: evaluating the mechanism of language and nonverbal cognitive function impairment based on neuroimage, exploring the role of nonverbal cognitive dysfunction in pathology and recovery mechanisms of patients with PSA, examining the therapeutic mechanism of non-invasive brain stimulation and electronic tele-rehabilitation equipment, and revealing not only the frontal and temporal areas but also brain network supporting nonverbal cognition. ConclusionIn recent years, some achievements have been made in the researches on verbal and nonverbal cognitive function, particularly in the studies of the neuroimaging-based mechanisms of common damage and the intervention effects of synergistic therapy. In the future, academic cooperation between different regions and different teams needs to be strengthened. Further exploration should be focused on refining the cognitive neuropsychology and exploring new technologies to improve the therapeutic effect of PSA.

Review of risk evaluation scores for benign end stage liver diseases recipients / 中华外科杂志

Liver transplant is an unreplaceable method for benign end-stage liver disease. The risk evaluation for the waiting list recipients and for post-transplant survival could provide practical indication for organ allocation. In recent years, there are two major kinds of evaluation scores. The first kind of evaluation scores is based on model for end-stage liver disease(MELD) score,including SOFT/P-SOFT score,UCLA-FRS score and BAR score. The other evaluation system is based on the concept of acute-on-chronic liver failure,including CLIF-C-ACLF score,TAM score,AARC-ACLF score and COSSH-ACLF score. The scores based on ACLF have been shown superior power in predicting waiting list survival and post-transplant prognosis than MELD. This article reviews the two kinds of evaluation scores,aiming for the better allocation policy and the better prognosis of benign end-stage liver disease.

Review of risk evaluation scores for benign end stage liver diseases recipients / 中华外科杂志

Liver transplant is an unreplaceable method for benign end-stage liver disease. The risk evaluation for the waiting list recipients and for post-transplant survival could provide practical indication for organ allocation. In recent years, there are two major kinds of evaluation scores. The first kind of evaluation scores is based on model for end-stage liver disease(MELD) score,including SOFT/P-SOFT score,UCLA-FRS score and BAR score. The other evaluation system is based on the concept of acute-on-chronic liver failure,including CLIF-C-ACLF score,TAM score,AARC-ACLF score and COSSH-ACLF score. The scores based on ACLF have been shown superior power in predicting waiting list survival and post-transplant prognosis than MELD. This article reviews the two kinds of evaluation scores,aiming for the better allocation policy and the better prognosis of benign end-stage liver disease.

Clinical efficacy of intra-articular injection with Triamcinolone acetonide in patients with juvenile idiopathic arthritis / 中华实用儿科临床杂志

Objective:To observe the clinical efficacy of intra-articular injection with Triamcinolone acetonide on the treatment of juvenile idiopathic arthritis (JIA).Methods:The clinical data of 26 children diagnosed with JIA undergoing the intra-articular injection of Triamcinolone acetonide for the joints with obvious swelling and pain at the Children′s Hospital Affiliated to Capital Institute of Pediatrics from October 2018 to December 2019 who were retrospectively analyzed.Erythrocyte sedimentation rate (ESR) and C-reactive protein(CRP) were tested before and after the application of Triamcinolone acetonide.Detailed clinical manifestations were recorded.The nonparametric Kruskal- Wallis test was used to compare the differences in clinical evaluation indicators and changes in laboratory tests at diffe-rent treatment times. Results:Among the 26 children, 8 were boys and 18 were girls.After the intra-articular injection of Triamcinolone acetonide, 9 cases (34.62%) achieved complete remission, 15 cases(57.69%) achieved partial remission, and 2 cases (7.69%) were not responsive to the intra-articular injection.The overall therapeutic efficacy was 92.31%.Compared with pre-treatment period, from 4 weeks after treatment, assessment of disease activity by the physicians and parents of the children was significantly improved after 4-week treatment, and the number of active joints, ESR and CRP and the Juvenile Arthritis Disease Activity Score with 27 joints (JADAS 27) gradually decreased, and the differences were statistically significant (all P<0.05). No adverse drug reactions were seen during the treatment and follow-up period. Conclusions:Intra-articular injection of Triamcinolone acetonide is effective in contro-lling joint symptoms of JIA with less adverse events.

Identification and Analysis of Complete Genomic Sequences of Soybean Mosaic Virus Isolates from Atractylodes macrocephala Koidz / 中国生物化学与分子生物学报

Soybean mosaic virus (SMV) is a critical pathogen that reduces the soybean yield and seed quality worldwide, and SMV has a restricted natural host range. In this study, we used sequence-independent amplification (SIA) methods to identify the viruses that may cause the Atractylodes macrocephala Koidz disease. Results revealed that there is SMV in diseased Atractylodes macrocephala leaves, and we named this isolate as SMV-Am. To further characterize the genomic structural and phylogenetic relationships of SMV-Am, genomic dsRNA was extracted, and the genomic sequence was amplified by RT-PCR and RACE. The results of bioinformatics analysis showed that the genomic RNA of SMV-Am is 9587 nucleotides in length, which conforms to the typical characteristics of Potyvirus. According to the sequence alignment of complete nucleotide sequences, SMV-Am showed the highest level of nucleotide homology and amino acid sequences to SMV-Liaoning, 96. 57% and 98. 86%, respectively. In addition, phylogenetic analysis based on the nucleotide sequences of other SMV isolates of SMV-Am revealed that SMV-Am was most closely related to SMV-Liaoning. Then, the SMV-Am protein was further analyzed by I-TASSER and PyMOL software, revealing that the key amino acid mutations led to the structural changes of P1, HC-Pro, P3, 6K2, NIa-pro and NIb proteins, with P1 the most obvious. Finally, recombination has been detected at the position of 6560-8950 nucleotides, the main parent is the isolate SMV-XFQ012 (accession number KP710875. 1), and the secondary parent is isolate SMV-pCB301-SC15 (accession number MH919386. 1). This study is the first report of SMV in Atractylodes macrocephala Koidz, and these results are expected to provide a scientific basis for the prevention and control of SMV infection on Atractylodes macrocephala Koidz.

Surgical treatment for aortic periannular abscess / 中国胸心血管外科临床杂志

@#Objective    To summarize the clinical data and efficacy of surgical treatment of aortic periannular abscess. Methods    The clinical data of 35 aortic periannular abscess patients admitted to our hospital from January 2009 to June 2019 were retrospectively analyzed, including 21 males and 14 females, aged 36 to 67 (53.0±12.3) years. Among them, there were 14 patients of native aortic valve endocarditis and 21 patients of prosthetic valve endocarditis (16 patients of mechanical valve and 5 patients of biological valve). Preoperative blood cultures were positive in 15 patients, including 8 patients of Staphylococcus aureus, 2 patients of Staphylococcus epidermidis, 3 patients of Streptococcus grass green, 1 patient of Pseudomonas aeruginosa, and 1 patient of Enterococcus. Results    Eleven patients underwent emergency or urgent surgery. Thirty patients underwent aortic valve replacement, and 5 patients underwent modified Cabrol surgery to replace the aortic root. Early postoperative complications included 1 patient of bleeding, 8 patients of low cardiac output syndrome, 5 patients of renal insufficiency, 10 patients of respiratory insufficiency, 3 patients of tracheotomy, 8 patients of pulmonary infection and 1 patient cerebrovascular accident. The postoperative follow-up period was 6 to 120 (53.6±20.8) months. During the follow-up, 4 patients died and 4 patients were lost. No infection recurred during the follow-up. Perivalval leakage occurred in 3 patients, and one patient underwent occlusion 12 months following the procedure. The survival curve indicated that the 1-year survival rate was 85.5%, and the 5-year survival rate was 67.3%. Conclusion    Although the lesions of periannular abscesses are complicated and critical, effective perioperative antibiotic treatment, individualized surgical timing, and appropriate surgical strategies can significantly reduce mortality and achieve better results.

Finite element analyses of retention of removable partial denture circumferential clasps manufactured by selective laser melting / 北京大学学报(医学版)

OBJECTIVE@#To compare the retentions of different designs of cobalt-chromium (Co-Cr), pure titanium (CP Ti), and titanium alloy (Ti-6Al-4V) removable partial denture (RPD) circumferential clasps manufactured by selective laser melting (SLM) and to analyze the stress distribution of these clasps during the removal from abutment teeth.@*METHODS@#Clasps with clasp arm size A (1.9 mm width/1.1 mm thickness at the body and 0.8-taper) or B (1.2 times A) and 0.25 mm or 0.50 mm undercut engagement were modeled on a prepared first premolar die, named as designs A1, A2, A3, and A4, respectively. The density and elastic modulus of SLM-built Co-Cr, CP Ti, and Ti-6Al-4V were measured and given to different groups of clasps. The density, elastic modulus, and Poisson ' s ratio of enamel were given to the die. The control group was the cast Co-Cr clasp with design A1, to which the density and elastic modulus of cast Co-Cr alloy were given. The Poisson's ratio of all metals was 0.33. The initial 5 N dislodging force was applied, and the maximum displacement of the clasp along the insertion path was computed. The load was reapplied with an increment of 5 N than in the last simulation until the clasp was completely dislodged. The retentive force range of different groups of clasps was obtained. The retentive forces of the SLM-built Co-Cr, CP Ti, and Ti-6Al-4V clasps with equivalent computed retentive force range to the control group were validated through the insertion/removal experiment. The von Mises stress distributions of these three groups of SLM-built clasps under 15 N loads were analyzed.@*RESULTS@#SLM-built Co-Cr, CP Ti, and Ti-6Al-4V clasps with designs B1 or B2, and Co-Cr clasps with design A2 had higher retentive forces than those of the control group. SLM-built CP Ti and Ti-6Al-4V clasps with design A1 had lower retentive forces than those of the control group. SLM-built Co-Cr clasp with design A1 and SLM-built CP Ti and Ti-6Al-4V clasps with design A2 had equivalent retentive forces to those of the control group. The insertion/removal experiment showed that the measured retentive forces of these three groups of SLM-built clasps were (21.57±5.41) N, (19.75±4.47) N, and (19.32±2.04) N, respectively. No statistically significant measured retentive force difference was found among these three groups of SLM-built clasps (P>0.05). The maximum von Mises stress of these three groups of SLM-built clasps exceeded their responding yield strength except for the Ti-6Al-4V one.@*CONCLUSION@#SLM-built Co-Cr circumferential clasps had higher retention than CP Ti and Ti-6Al-4V ones with the same clasp arm size and undercut engagement. The retention of SLM-built circumferential clasps could be adjusted by changing the undercut engagement and clasp arm size. If SLM-built circumferential clasps are used in clinical practice, the Ti-6Al-4V clasp with clasp arm size A and 0.50 mm undercut engagement is recommended considering the long-term use of RPD in the patient's mouth.

Research Progress of Exosomal microRNA in Cardiovascular Disease and Its Forensic Application Prospects / 法医学杂志

Exosomal microRNAs (miRNAs) are miRNAs that are mediated by exosomes to achieve cell-to-cell communication, and they are widespread in organisms. In recent years, the key role of the multiple biological functions of exosomal miRNAs in the occurrence and development of cardiovascular diseases has been confirmed by a large number of studies, which has become a hot spot in clinical and basic research. Sudden cardiac death caused by cardiovascular disease is one of the important contents in forensic medical identification. This article introduces the research progress of cardiovascular disease prediction, treatment and prognosis on exosomal miRNA. The prospects of the application in forensic medical identification are discussed.

Blood flow restriction training can enhance the lower limb joint functioning of persons with rheumatoid arthritis / 中华物理医学与康复杂志

Objective:To investigate the effect of blood flow restriction training (BFRT) on the rehabilitation of lower limb joint function in persons with rheumatoid arthritis (RA).Methods:Sixty RA patients were randomly divided into an observation group and a control group, each of 30. Both groups were given conventional slow-acting anti-rheumatic medication with low-dose glucocorticoids. Everyone received basic exercise training including joint training and plyometric training, but the observation group additionally received BFRT training during which constant pressure in an inguinal cuff restricted blood flow to the lower limbs. The observation group′s training program also included double leg push-ups and double knee extensions. Before, and 12 weeks after the intervention, the joint symptoms were assessed using swollen joint counts out of 28 joints (SJC28), tender joint counts out of 28 joints (TJC28) and pain reported using a visual analogue scale (VAS). Lower limb motor functioning was assessed using a timed-stands test (TST) and the timed-up-and-go (TUG) test. Life quality was evaluated using a health assessment questionnaire (HAQ) and arthritis self-efficacy scale-8 (ASES-8). Disease activity was quantified using a clinical disease activity index (CDAI), a simplified disease activity index (SDAI) and disease activity scoring with a 28-joint count (DAS28).Results:There were no significant differences between the two groups before the intervention. After the intervention all of the indicators except the DAS28 had improved significantly in the observation group. At that point they were also all better than the control group′s averages, where no significant improvement was observed. No serious adverse events occurred in either group during the experiment.Conclusions:BFRT can safely relieve joint symptoms and improve the lower limb motor functioning and life quality of RA sufferers. It is worthy of further clinical research and promotion.

Rapid Enrichment and Activity Research of Antithrombotic Proteins from Pheretima vulgaris / 中国实验方剂学杂志

Objective:A strong antithrombotic protein component， named PvQ， was purified and enriched from total protein of <italic>Pheretima vulgaris</italic>，<italic> </italic>a<italic> </italic>traditional Chinese medicine. Moreover， we evaluated its fibrinolytic and anticoagulant activity， and expected to provide reference for the research on antithrombotic substances of Pheretima. Method:A rapid <italic>in</italic> <italic>vitro</italic> activity-oriented separation combined with the AKTA-Pure protein purification system conducted on <italic>P. vulgaris</italic>. Meanwhile， the fibrinolytic and anticoagulant activities of PvQ were measured by fibrin plate method and fibrinogen-thrombin time （Fibg-TT） method. And the <italic>in vitro</italic> thrombolysis assay was used for evaluating the lysis ability of PvQ to thrombus. Then the stability of PvQ was also analyzed for its anticoagulant activity at different pH and temperature. Result:The PvQ was successfully enriched and its activity was determined to have significant fibrinolytic and anticoagulant activities. And the result of <italic>in vitro</italic> thrombolysis assay revealed that PvQ could hydrolyze more than 80% of thrombus after 5 h of incubation at 37 ℃. In addition， the changes of temperature and pH had significant effects on antithrombotic activity， and this study showed that PvQ was rapidly inactivated at ≥60 ℃ or in acidic conditions （pH<7）. While， the activity of PvQ was unaffected or less affected at ≤50 ℃ and under alkaline conditions. Conclusion:A feasible preparation method of PvQ is established， and it can affect fibrin and fibrinogen at the same time， thus exerting a dual fibrinolytic effect and possessing significant fibrinolytic and anticoagulant activities. It provides a scientific interpretation for the treatment of thrombotic diseases by PvQ and a reference for the development of antithrombotic protein products of Pheretima.

Rac1 Signaling in Amygdala Astrocytes Regulates Fear Memory Acquisition and Retrieval / 神经科学通报·英文版

The importance of astrocytes in behavior control is increasingly appreciated, but little is known about the effects of their dynamic activity in regulating learning and memory. In the present study, we constructed AAVs of photoactivatable and photoinactivatable Ras-related C3 botulinum toxin substrate 1 (Rac1) under the mGFAP promoter, which enabled the manipulation of Rac1 activity in astrocytes by optical stimulation in free-moving mice. We found that both up-regulation and down-regulation of astrocytic Rac1 activity in the basolateral amygdala (BLA) attenuated memory acquisition in a fear conditioning mouse model. Meanwhile, neuronal activation in the BLA induced by memory acquisition was inhibited under both the up- and down-regulation of astrocytic Rac1 activity during training. In terms of the impact on fear memory retrieval, we found both up- and down-regulation of BLA astrocytic Rac1 activity impaired memory retrieval of fear conditioning and memory retrieval-induced neuronal activation. Notably, the effect of astrocytic Rac1 on memory retrieval was reversible. Our results demonstrate that the normal activity of astrocytic Rac1 is necessary for the activation of neurons and memory formation. Both activation and inactivation of astrocytic Rac1 activity in the BLA reduced the excitability of neurons, and thereby impaired fear memory acquisition and retrieval.

Establishment and Optimization of a Method for Rapid Extraction of Plant Virus RNA Based on Functional Magnetic Beads / 中国生物化学与分子生物学报

Plant virus diseases are one of the major diseases restricting erop production.Timely identification of their pathogen and development rules is the prerequisite for effective control of their large- scale spread.However, long cycle, tedious steps and strict detection environment were the disadvantages existing in the detection technology of plant virus disease.In this study, Tobacco Mosaic virus (TMV) was used as a model to he extract UNA based on CMBs-ACPtmv , which was design based on the principle of complementary base pairing.Meanwhile, the experimental conditions were optimized and analyzed, including the preparation conditions of functionalized magnetic beads, the reaction conditions during extraction, and the sensitivity, stability and other properties of the method.The results showed the ability to capture RNA of CMBs-ACPtmv were best when prepared with 4 fxmol capture probe (ACPTMV ) and 0.08 mg carboxyl magnetic beads (CMBs) ; After 3 min of extraction, CMBs-ACPtmv has the best RNA extraction effect, but when the extraction temperature of CMBs-ACPtmv was changed, its extraction capacity showed no significant change; In the comprehensive performance evaluation, the sensitivity of CMBs-ACPjjjv can reach 2.5 ng/fxL, and the detection stability is good.Compared with conventional RNA extraction technology, CMBs-ACPimv has outstanding advantages in detection time and sample consumption.The functional magnetic beads extraction method established in this study is fast, safe and simple.It can achieve rapid extraction of plant virus RNA with simple equipment, which has a broad application prospect.

Rehmanniae Radix and Rehmanniae Radix Praeparata improve diabetes induced by high-fat diet coupled with streptozotocin in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway / 中国中药杂志

This study investigated the differential mechanisms of Rehmanniae Radix and Rehmanniae Radix Praeparata in improving diabetes in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway. The diabetic mouse model was established with high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days), after which the mice were randomly divided into model group, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, catalpol group(250 mg·kg~(-1)), 5-hydroxymethylfurfural(5-HMF) group(250 mg·kg~(-1)), metformin group(250 mg·kg~(-1)), with the normal group also set. The organ indexes of heart,liver, spleen, lung, kidney and pancreas were calculated after four weeks of administration. The pathological changes and fibrosis of pancreas, kidney and liver in mice were observed by hematoxylin-eosin(HE) staining and Masson staining. Western blot was used to determine the expression levels of Toll-like receptor-4(TLR4), nuclear factor-κB(NF-κB), Nod-like receptor protein 3(NLRP3),interleukin-1β(IL-1β), adenosine monophosphate-activated protein kinase(AMPK), phosphorylated AMPK(p-AMPK) in the pancreas, kidney and liver of mice. Compared with the model group, the administration groups witnessed significant decrease in the liver,spleen, kidney, pancreas and fat indexes of diabetic mice, and there was no significant difference in heart and lung indexes. The pathological states and fibrosis of pancreatic, kidney and liver tissues were significantly improved after administration. Additionally, the expression levels of TLR4, NF-κB and NLRP3 in pancreas, kidney and liver of diabetic mice were significantly lowered. The expression levels of p-AMPK/AMPK were enhanced significantly in kidney and liver of mice in Rehmanniae Radix group while in pancreas, kidney and liver in Rehmanniae Radix Praeparata group. This suggests that Rehmanniae Radix and Rehmanniae Radix Praeparata differ in the mechanism of regulating energy metabolism of multiple organs and thereby exerting anti-inflammatory effects to alleviate symptoms of diabetic mice.

Related factors of ophthalmoplegia in children with ocular myasthenia gravis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the related-factors analysis of extra-ocular muscle paralysis in pediatric ocular myasthenia gravis(OMG). <p>METHODS: A retrospective, observational cohort study was performed of 203 pediatric patients diagnosed with OMG between Nov.2011 and May 2020 at Wuhan Children's Hospital. Data on clinical features and laboratory indicators of children with OMG were statistically analyzed between extra-ocular muscle paralysis group and non-extra-ocular muscle paralysis group, which were classified by pediatric ophthalmologists based on ocular manifestations. The Logistic regression analysis was performed immediately after that to determine the independent factors. <p>RESULTS: Totally 203 pediatric OMG, 97(47.8%)had extra-ocular muscle paralysis with various clinical features including strabismus(<i>n</i>=69, 71.1%), torticollis(<i>n</i>=18, 18.6%). Among the 97 children, 79(81.4%)patients were presented with monocular involvement, of which 53(54.6%)had single extra-ocular muscle paralysis. The most common was medial rectus muscle(<i>n</i>=19, 35.8%). Between extra-ocular muscle paralysis group and non-extra-ocular muscle paralysis group, there were statistically significant difference(<i>P</i><0.05)in age, serum immunoglobulin M(IgM), serum free triiodothyronine(FT3), serum thyroglobulin, and glucocorticoid combination pyridostigmine bromide therapy(72.2% <i>vs</i> 38.7%). Furthermore, the level of FT3(<i>OR</i>=2.006, 95%<i>CI</i>:<i> </i>1.233-3.263)and glucocorticoid combined treatment(<i>OR</i>=4.328, 95%<i>CI</i>: 1.936-9.677)were the related-factors affecting the extra-ocular muscle paralysis of children with ocular myasthenia gravis. <p>CONCLUSION: Extra-ocular muscle paralysis was the common ocular manifestation in pediatric OMG. Monocular involvement was more common than binocular involvement especially the medial rectus muscle, while diplopia was rare. The level of FT3 can be considered as an important immune indicator to evaluate extra-ocular muscle paralysis in children with ocular myasthenia gravis.