Functional and Structural Analysis of a Novel Acyltransferase from Pathogenic Phytophthora melonis.

This investigation characterizes an acyltransferase enzyme responsible for the pathogenicity of Phytophthora melonis. The protein was characterized in vitro for its physicochemical properties. The biochemical characterization, including thermal and pH stability, revealed the 35 °C temperature and 7.0 pH as the optimum conditions for the enzyme. Applying the Tween-80 solution enhanced the activity up to 124.9%. Comprehensive structural annotation revealed two domains, A (ranging from residues 260 to 620) and B (ranging from 141 to 219). Domain A had transglutaminase (T-Gase) elicitor properties, while B possessed antifreeze features. Rigorous sequence characterization of the acyltransferase tagged it as a low-temperature-resistant protein. Further, the taxonomic distribution analysis of the protein highlighted three genera in Oomycetes, i.e., Pythium, Phytophthora, and Plasmopara, bearing this protein. However, some taxonomic groups other than Oomycetes (i.e., archaea and bacteria) also contained the protein. Functional studies of structurally analogous proteins spanned 10 different taxonomic groups. These revealed TGase elicitors (10%), phytopathogen effector proteins RxLR (4%), transporter family proteins (3%), and endonucleases (1%). Other analogues having one percent of their individual share were HIV tat-specific factor 1, protocadherin fat 4, transcription factor 1, and 3-hydroxyisobutyrate dehydrogenase. Because the plant infection by P. melonis is a complex process regulated by a profusion of extracellular signals secreted by both host plants and the pathogen, this study will be of help in interpreting the cross-talk in the host-pathogen system.

Early Improvement of Acute Respiratory Distress Syndrome in Patients with COVID-19: Insights from the Data of ICU Patients in Chongqing, China

Acute respiratory distress syndrome (ARDS) may be the main cause of death in patients with coronavirus disease 2019 (COVID-19). Herein, we retrospect clinical features, outcomes and ARDS characteristics of 75 intensive care unit (ICU) patients with COVID-19 in Chongqing, China. We found a 5.3% case fatality rate of the ICU patients in Chongqing. 93% patients developed ARDS during the intensive care, and more than half were moderate. However, most of the patients (55%) supported with high flow nasal cannula (HFNC) oxygen therapy, but not mechanical ventilation. Nearly one third of patients with ARDS got an early improvement (eiARDS), and the rate is much higher than the other causes of ARDS in a previous study. Patients with eiARDS had a higher survival rate and lower length of ICU stay. The age (< 55 years) is an independent predictor for the eiARDS, and stratification of COVID-19 patients by age is recommended.

Metabolic and Proteomic Perspectives of Augmentation of Nutritional Contents and Plant Defense in Vigna unguiculata.

The current study enlists metabolites of Alstonia scholaris with bioactivities, and the most active compound, 3-(1-methylpyrrolidin-2-yl) pyridine, was selected against Macrophomina phaseolina. Appraisal of the Alstonia metabolites identified the 3-(1-methylpyrrolidin-2-yl) pyridine as a bioactive compound which elevated vitamins and nutritional contents of Vigna unguiculata up to ≥18%, and other physiological parameters up to 28.9%. The bioactive compound (0.1%) upregulated key defense genes, shifted defense metabolism from salicylic acid to jasmonic acid, and induced glucanase enzymes for improved defenses. The structural studies categorized four glucanase-isozymes under beta-glycanases falling in (Trans) glycosidases with TIM beta/alpha-barrel fold. The study determined key-protein factors (Q9SAJ4) for elevated nutritional contents, along with its structural and functional mechanisms, as well as interactions with other loci. The nicotine-docked Q9SAJ4 protein showed a 200% elevated activity and interacted with AT1G79550.2, AT1G12900.1, AT1G13440.1, AT3G04120.1, and AT3G26650.1 loci to ramp up the metabolic processes. Furthermore, the study emphasizes the physiological mechanism involved in the enrichment of the nutritional contents of V. unguiculata. Metabolic studies concluded that increased melibiose and glucose 6-phosphate contents, accompanied by reduced trehalose (-0.9-fold), with sugar drifts to downstream pyruvate biosynthesis and acetyl Co-A metabolism mainly triggered nutritional contents. Hydrogen bonding at residues G.357, G.380, and G.381 docked nicotine with Q9SAJ4 and transformed its bilobed structure for easy exposure toward substrate molecules. The current study augments the nutritional value of edible stuff and supports agriculture-based country economies.

Effect of spinal cord decellularized scaffold on the repair of spinal cord defects in rats / 解剖学报

Objective Exploring the effect of spinal cord decellularized scaffold on spinal cord defects and observing the behavior and regeneration of rats after operation. Methods The spinal cords of 30 SD rats were treated with 3% Triton X-100 and 2% sodium deoxycholate on oscillator. The cell residue and the spatial structure of the tissue were compared before and after treatment, in order to understand the tissue structure of the stent itself. 90 SD rats were randomly divided into control group, simple injury group and stent transplantation group. Excision of the spinal cord 9-10 segments in the simple injury group and the stent graft group the acellular scaffold was transplanted to the stent graft group. Behavioral scores were observed postoperatively. At 4, 8, and 12 weeks, the spinal cords of the injured part of the rats were taken for HE staining and immunofluorescence detection of nerve regeneration-related proteins. Results After decellularization of the spinal cord, the nerve cells and axons were completely removed, and the extracellular matrix of the spinal cord was preserved. Scanning electron microscopy revealed that the scaffold retained a certain porous network scaffold structure. In the experiment of decellularized scaffold in vivo, the Basso-Beattie-Bresnahan(BBB) score showed that the recovery of hindlimb motor function in rats with decellularized scaffolds was better than that in rats with simple injury. HE staining showed that the decellularized scaffold could fill the defect of the spinal cord segment and accelerate the repair process of the injured spinal cord. Immunofluorescence showed that there was a certain axonal regeneration in the injured part of the stent transplantation group. Conclusion The spinal cord decellularized scaffold retains the extracellular matrix and has a certain spatial structure, which can accelerate the process of spinal cord defect repair to a certain extent, and has a certain promoting effect on nerve regeneration.

Benzenedicarboxylic acid upregulates O48814 and Q9FJQ8 for improved nutritional contents of tomato and low risk of fungal attack.

BACKGROUND: Tomato is an important food item and a cocktail of phytonutrients. In the current study, metabolites from a non-pathogenic fungal species Penicillium oxalicum have been exploited to obtain nutritionally augmented tomato fruits from the plants to better withstand against Alternaria alternata infection. RESULTS: Initially, bioactivity-guided assay and chromatographic analyses identified the bioactive metabolites of P. oxalicum [benzenedicarboxylic acid (BDA) and benzimidazole]. Then, ≥3 times elevated quantities of vitamins and other nutritional elements (protein, fat, fibers, and carbohydrates) were achieved by the foliar application of BDA. The maximum increase (625.81%) was recorded in riboflavin contents; however, thiamine showed the second highest enhancement (542.86%). Plant metabolites analysis revealed that jasmonic acid contents were boosted 121.53% to significantly enhance guaiacyl lignin defenses along with the reduction in coumarin contents. The protein profile analysis explored three most actively responding protein species toward BDA applications, (i) palmitoyltransferase protein Q9FLM3; (ii) serine/threonine-protein kinase O48814; and (iii) E3 ubiquitin-protein ligase Q9FJQ8. The O48814 improved plant defenses; whereas, Q9FJQ8 protein was negatively regulating cysteine-type endopeptidase activity and assisted plant to resist schedule alterations. Tomato cultivar with more active innate metabolism was found to be more responsive toward BDA. Furthermore, the bioactive compounds were enriched by using the two-step extraction method of ethyl acetate and chloroform, respectively. CONCLUSION: Penicillium oxalicum a non-pathogenic fungal species, produced BDA, induced nutritional contents in tomato and protected it against Alternaria alternata. The current study is the first report on the bioactivity of BDA and benzimidazole concerning the nutritional enhancement and plant defense improvement. © 2019 Society of Chemical Industry.

Simultaneous determination of eight alkaloids and oleandrin in herbal cosmetics by dispersive solid-phase extraction coupled with ultra high performance liquid chromatography and tandem mass spectrometry.

We utilized ultra-high performance liquid chromatography with tandem mass spectrometry and dispersive solid-phase extraction to develop a new method for the detection of nine analytes (scopolamine, cephaeline, strychnine, hyoscyamine, brucine, hydrastine, ajmalicine, colchicine, and oleandrin) in herbal cosmetics. Acetonitrile/water and 2-propylaminoethylamine were used to disperse and purify during the dispersive solid-phase extraction step. The analytes were separated by a Waters UPLC HSS T3 column and detected through electrospray ionization source in the positive mode with multi-reaction monitoring conditions. Under the optimal conditions, the calibration curves were linear in the range of 0.2-100.0 µg/L with the correlation coefficients higher than 0.995. The method limit of quantitation (S/N = 10) were 5.0 µg/kg for oleandrin and 1.0 µg/kg for the other eight alkaloids. The mean recoveries at three spiked concentration levels of 1.0-10.0 µg/kg were in the range of 86.9-116.5% with the intra-day relative standard deviations (n = 6) ranging from 2.4 to 8.8%, and inter-day relative standard deviations ranging from 2.7 to 5.7%. This method is accurate, simple and rapid, and has been applied to the quality supervision of herbal cosmetics in Guangzhou.

Automated hollow-fiber liquid-phase microextraction coupled with liquid chromatography/tandem mass spectrometry for the analysis of aflatoxin M1 in milk.

An automated hollow fiber liquid-phase microextraction (HF-LPME) coupled with liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed for the extraction and determination of aflatoxin M1 (AFM1) in milk samples. Parameters affecting the extraction efficiency, such as the extraction phase, matrix conditions, extraction time and temperature, were investigated. Under the optimal conditions (ratio of water to milk, 4:1; extraction time, 50 min; extraction temperature, 50°C; extraction phase, 50 mg L(-1) anti-AFM1 antibody in PBS buffer solution; volume of HCl solution, 250 µL; agitation speed, 250 rpm), the matrix-matched calibration curve for AFM1 determination showed good linearity in the range of 0.25-5 µg kg(-1). The enrichment factor (EF) reached 48, and the limits of detection and quantification were 0.06 and 0.21 µg kg(-1), respectively. The developed method was successfully applied for the extraction of AFM1 from spiked milk samples, with recoveries from 61.0% to 106.7%. The method was highly specific to AFM1 analysis, and the results demonstrated that the method can be automated, inexpensive, and free from interference.

Delayed union or nonunion of the ulna after intramedullary nailing for pediatric forearm fractures / 中国骨伤

<p><b>OBJECTIVE</b>To analyze the causes of delayed union or nonunion of the ulna after intramedullary nailing in pediatric forearm fractures.</p><p><b>METHODS</b>From February 2005 to February 2010,5 patients with forearm fractures who were treated with titanium elastic nailing (TEN) were identified to fulfill the criteria of having developed a delayed union or nonunion of the ulna. The causes of delayed union or nonunion were investigated according to mechanism of injury, fracture location, treatments methods and postoperative management. All patients were male and the age was 3 to 14 years old with an average of 9.4 years. All fractures were located on the mid-third part of forearm. Two cases had a re-fracture. Among them, 3 cases caused by high-energy injury and 2 cases by falling down. Open reduction were performed in 4 cases while the other one was treated with closed reduction. Four patients were immobilized in an above-elbow cast, postoperatively.</p><p><b>RESULTS</b>All patients were followed up from 7 to 19 months with an average of 11.4 months. There were 4 delayed union and 1 nonunion. Three patients healed after the removal of the nail and avoidance of weight-bearing. Two patients healed by replacing another fixation. No patients had soft-tissue irritation or nail-entry-site infections.. The clinical effect was evaluated according to Daruwalla and Price scores with 3 excellent and 2 good of the results.</p><p><b>CONCLUSIONS</b>Using titanium elastic nailing for the treatment of pediatric both-bone forearm fractures is a good method. However,strict indication selection should be followed to avoid delayed union or nonunion.</p>

Clinical analysis of retroperitoneal laparoscopic renal adipose capsule outer pathway in the treatment of chyluria / 重庆医学

Objective To evaluate the surgical techniques and clinical efficacy of retroperitoneoscopic ligation of renal lymphatic vessels outside adipose capsule and inside adipose capsule for comparison in the treatment of chyluria.Methods Retrospective anal-ysis of the retroperitoneal laparoscopic adipose capsule of kidney pedicle lymphatic duct ligation(group Ⅰ)and adipose capsule pathways in the kidney totally or subtotally free renal pedicle lymphatic ligation(group Ⅱ)and 1 1 1 patients with chyluria clinical data,compared two groups of operation time,bleeding volume,the rate of analgesia,postoperative gastrointestinal function recovery time,drainage time,postoperative recovery time,postoperative hospital stay and postoperative complications and other index differ-ence.Results Retroperitoneal laparoscopic operation group of adipose capsule in operation time,bleeding volume,postoperative an-algesia,postoperative gastrointestinal function recovery time,drainage time,postoperative recovery time,postoperative hospital stay and postoperative complications were better than the adipose capsule of kidney totally or subtotally free operation group,the differ-ence was statistically significant(P<0.05).Conclusion Retroperitoneal laparoscopic renal adipose capsule of kidney pedicle lym-phatic disconnection for the treatment of chyluria effect,and adipose capsule operation ways,the method less trauma,quicker recov-ery,can completely replace the renal adipose capsule total or Sub-total of free kidney pedicle lymphatic disconnection operation,wor-thy of clinical application.

Advance in study on pharmacological effect of Eucommiae Folium / 中国中药杂志

Eucommia ulmoides is a valuable traditional Chinese medicine, whose cortexes have long been used as medi cines. Due to the scarcity of its resources, people began using its leaves instead of cortexes in medicines. Eucommiae Folium and its leaves have many pharmacological effects and thereby being clinically applied as genuine traditional Chinese medicines. Modern pharmological studies have showed that Eucommiae Folium leaves have such effects as blood pressure reduction, blood lipid regulation, cardiovascular protection, anti-obesity, anti-inflammation, anti-virus, enhancement of immunologic function, resistance against senility and anti-fatigue. In clinic, Eucommiae Folium is mainly used to treat hypertention and obstetrical and gynecological disease. The essay summarizes the latest advance in domestic and foreign studie on pharmacological effeets and clinical applications of Eucommiae Folium leaves, and thus providing reference for studies on new drugs of Eucommiae Folium leave.

An analysis of randomized control trials articles published from 2004-2009 in Chinese Journal of Hepatobiliary Surgery / 中华肝胆外科杂志

Objective To study the academic level of randomized control trials(RCTs)published from 2004 to 2009 in Chinese Journal of Hepatobiliary Surgery.Methods Published work about RCTs in 72 issues of Chinese Journal of Hepatobiliary Surgery was searched by finding relevant articles published during 2004-2009.Correlated indexes including literature received time, literature publishing time, the total number of citations, number of citations in Chinese, number of citations in English, research funding situations, the number of authors, article affiliations in all RCTs, together with single(or multiple)center study situation, positive conclusions, the starting time, the end time,and number of patients recruited in clinical RCTs were extracted for aggregate analysis.Results During the past six years, a total of 135 articles of RCT were published in Chinese Journal of Hepatobiliary Surgery, accounting for 8.7% in all clinical studies.The average number of RCTs in each issue was 1.88.The average time for publishing time delay was 450 days.The total number of citations in RCTs was 1252 and the average number of RCTs for citations was 9.3.53 RCT studies were financed, accounting for 39% of the total number of research papers.Beijing Friendship Hospital attached to the Capital University of Medical Sciences had published 8 papers of RCT and it is the institution of publishing the largest number of such papers in the journal.The number of multi-center study for RCTs was one and all RCT studies had significant conclusions.The average research duraton in RCTs was 45 months.The total number of patients in 33 clinical RCTs was 2705 and the average number of patients recruited in each RCT was 82.Conclusion Chinese Journal of Hepatobiliary Surgery put emphasis on the strongest evidence and the most valuable RCT researches that have great significance in the field of domestic hepatobiliary surgery for clinical treatment and experimental research.

An analysis of randomized controlled trials published in the Chinese Journal of Gastrointestinal Surgery between 2003-2009 / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the academic level of randomized controlled trials (RCT) published in the Chinese Journal of Gastrointestinal Surgery between 2003 and 2009.</p><p><b>METHODS</b>Published RCTs in the 42 issues of the Chinese Journal of Gastrointestinal Surgery was searched for relevant articles published between 2003 and 2009. Data extracted for analysis included the time at manuscript received, publication time, total number of citations, number of citations in Chinese, number of citations in English, author's affiliations, single- or multi- center study, positive conclusions from RCTs, number of patients recruited in RCTs, research funding source, the start time, the finish time and the number of authors in RCTs.</p><p><b>RESULTS</b>During the past seven years, a total of 80 clinical RCTs were published in the Chinese Journal of Gastrointestinal Surgery, accounting for 12% of all the clinical studies published in the journal, and the average number of RCTs in each issue was 1.6. The average delay time before publication was 208 days. The total number of citations and the total number of patients in RCTs were 685 and 9402. The average number of citations, the average number of patients recruited in each RCT, and the average research period in RCTs were 8.6, 118 and 29.2. There were 7 multi-center studies, and the number of single-center study was 73. All the RCT studies had significant conclusions, and 17 (21.3%) RCT studies were funded. Nanjing general hospital of Nanjing military command had the largest number of RCTs (n=6).</p><p><b>CONCLUSION</b>The Chinese Journal of Gastrointestinal Surgery puts emphasis on clinical studies of high evidence level such as RCT, which provide evidence for making the clinical guidelines in the specialty of gastrointestinal surgery.</p>

The effect of hepatitis B virus X protein on the c-met promoter activity in HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To confirm the effect of hepatitis B virus X (HBx) protein on the c-met promoter activity in HepG2 cells.</p><p><b>METHOD</b>The expression of c-met protein was detected by western blot in HBx-transfected HepG2 cells, the human c-met promote activity was checked by luciferase assay using five different constructs with deletion or point mutation.</p><p><b>RESULTS</b>HBx protein stimulated the expression of the c-met in HepG2 cells. The enhanced expression of c-met in HBx-transfected cells was mediated by the activation of AP-2 and SP-1 transcriptional activity at the c-met promoter region (-183bp--100bp), HBx increased the invasiveness of HepG2 cells as determined by Matrigel invasion assay.</p><p><b>CONCLUSION</b>These results suggests that HBx induces the expression of c-met through the activation of AP-2 and SP-1 activity at the promoter region; in addition, our data indicate that HBx stimulates the invasive potential of HepG2 cells.</p>

Impaired Delta NP63 Expression is Associated with Poor Tumor Development in Transitional Cell Carcinoma of the Bladder

The oncogenic isoform of the p63 protein, delta NP63, plays an important role in the pathogenesis of many epithelial carcinomas, and emerging evidences suggest that delta NP63 is a promising drug target. However, the functions of delta NP63 in transitional cell carcinoma of bladder (TCCB) are poorly defined. In this study, a delta NP63 shRNA expression vector was transfected into TCCB cell line 5637 and cell cycling, cell proliferation and protein expression were assessed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dimethyl tetrazolium bromide (MTT) assay, and immunohistochemistry, respectively. The delta NP63 shRNA expression vector was also injected into 5637 cell xenograft tumors in nude mice, and tumor size was measured, tumor tissue morphology was assessed by immunohistopathology and transmission electron microscopy. In the in vitro study, delta NP63 shRNA transfection caused successful delta NP63 gene silencing and resulted in significant arrest of cell cycling and cellular proliferation (p<0.05) as well as cyclin D1 expression. In the nude mouse xenograft model, delta NP63 shRNA greatly inhibited tumor growth, induced tumor cell apoptosis (p<0.05) and resulted in cyclin D1 downregulation. Our data suggest that delta NP63 may play an oncogenic role in TCCB progression through promoting cell survival and proliferation. Intratumoral administration of delta NP63-specific shRNA suppressed tumor delta NP63 expression and cellular proliferation while promoted tumor cellular apoptosis, and therefore inhibited tumor growth and improved survival of xenograft-bearing mice, which was not accompanied by significant signs of systemic toxicity.

Prediction of the possible tertiary structure alterations of p53 protein following point mutation in p53 gene condon 282 in lung cancer cells / 南方医科大学学报

This study was carried out to predict the possible tertiary structure alterations of p53 protein after point mutation of p53 gene condon 282 in lung cancer cells based on the latest 3D structure analysis platform series of Phyre software. It was found that the p53 gene condon 282 mutation (Arg/Leu) may destabilize the H2 helix and DNA binding in the major groove by compromising the contacts of p53 protein with the beta-hairpin of DNA binding surface.

Antisense oligonucleotide targeting livin induces apoptosis of human renal carcinoma cell 786-O in vitro and its mechanism / 第三军医大学学报

Objective To investigate the effect of antisense oligonucleotides (ASODN) targeting livin on the inhibition of livin mRNA and protein expression and the apoptosis of human renal carcinoma cell line 786-O cells. Methods Specific phosphorothioate antisense oligodeoxynucleotides targeting livin were synthesized and then transfected into 786-O cells. The expressions of livin mRNA were detected by RT-PCR. Expression and location of livin protein were observed by confocal laser scanning microscope (CLSM). Apoptosis rate of 786-O cells was investigated by flow cytometer. The activity of Caspase-3 was detected by colorimetric assay. Results After the transfection of ASODN, the expression of livin mRNA was decreased (P

Isolation and identification of steroidal saponins in total saponin from Dioscorea nipponica Makino / 药学学报

<p><b>AIM</b>To investigate the water-soluble steroidal saponins in total saponin from Dioscorea nipponica Makino and look for new active compounds.</p><p><b>METHODS</b>The compounds were isolated with silica gel, PTLC and HPLC, and their structures were elucidated by acid hydrolysis, physical and chemical data and spectral analysis (IR, NMR, MS, HMQC, HMBC) as well as chemical correlations.</p><p><b>RESULTS</b>The two steroidal saponins (water-insoluble saponin and water-soluble saponin) were isolated from the total saponin of Dioscorea nipponica Makino. The structures were elucidated as diosgenin 3-O-[alpha-L-rhamnopy ranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]]-beta-D-glucopyranoside (I), diosgenin 3-O-[alpha-L-rhamnopyranosyl (1-->3)-alpha-L-rhamnopyranosyl (1-->4)-alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside (II).</p><p><b>CONCLUSION</b>Compound II is a new steroidal saponin and firstly isolated from Dioscorea nipponica Makino. It was named as dioscin Dc.</p>

Animal experimental study on the treatment of bladder transitional cell carcinoma by survivin-siRNA / 重庆医科大学学报

Objective:To construct the plasmid containing short hairpin RNA(shRNA)of survivin and modify human bladder transitional cell carcinoma(BTCC)T24 cell line with it to evaluate its inhibition on tumor growth and proliferation and further discuss its functions in T24 cells.Methods:Survivin specific oligonucleotides were designed and synthesized.These oligonucleotides were annealed to form double strand DNA fragment and this fragment was inserted into plasimid PTZU6+1 to generate the plasimid pshRNA-survivin2.Then T24 cells were inoculated into the right-back legs of BALB/c nude mice to establish bladder cancer model.Through plasimid PTZU6+1,T24/survivin-siRNA expressing survivin was successfully constructed to study the effects of survivin-siRNA on BTCC growth and proliferation.Microscope was used to observe the morphological changes.Histological examination was under HE staining.Survivin was detected by SABC to evaluate the expression of survivin after the treatment of survivin-siRNA through PTZU6+1.Results:The expression of survivin in transfected T24 cells in vivo was markedly depressed as compared with control groups(P