Depsides and depsidones have attracted attention for biosynthetic studies due to their broad biological activities and structural diversity. Previous structure‒activity relationships indicated that triple halogenated depsidones display the best anti-pathogenic activity. However, the gene cluster and the tailoring steps responsible for halogenated depsidone nornidulin ( 3) remain enigmatic. In this study, we disclosed the complete biosynthetic pathway of the halogenated depsidone through in vivo gene disruption, heterologous expression and in vitro biochemical experiments. We demonstrated an unusual depside skeleton biosynthesis process mediated by both highly-reducing polyketide synthase and non-reducing polyketide synthase, which is distinct from the common depside skeleton biosynthesis. This skeleton was subsequently modified by two in-cluster enzymes DepG and DepF for the ether bond formation and decarboxylation, respectively. In addition, the decarboxylase DepF exhibited substrate promiscuity for different scaffold substrates. Finally, and interestingly, we discovered a halogenase encoded remotely from the biosynthetic gene cluster, which catalyzes triple-halogenation to produce the active end product nornidulin ( 3). These discoveries provide new insights for further understanding the biosynthesis of depsidones and their derivatives.

OBJECTIVE To inv estigate the in vitro inhibitory effects of acteoside on cytochrome P 450（CYP）enzymes in liver microsomes of rats. METHODS Using probe substrates method ，acteoside（0.1，0.3，1，3，10，30 μmol/L）was incubated with probe substrates phenacetin ，mephentoin，diclofenac，coumarin，dextromethorphan and testosterone （substrates of CYP 1A2， CYP2C19，CYP2C9，CYP2A6，CYP2D6 and CYP 3A4 enzymes，respectively）in liver microsomes of rats. Another blank control group and positive inhibitor group [ α-naphthoflavone，ticlopidine，sulfabendazole，pilocarpine，quinidine and ketoconazole （inhibitors of CYP 1A2，CYP2C19，CYP2C9，CYP2A6，CYP2D6 and CYP 3A4 enzymes，respectively）] were set up. Using indapamide as the internal standard ， the contents of corresponding metabolites （acetaminophen， 4-hydroxymephenytoin， 4-hydroxydiclofenac，7-hydroxycoumarin，dextran，6 β-hydroxytestosterone） were detected by ultra high performance liquid chromatography-tandem mass spectrometry . The IC 50 values were calculated by GraphPad v 8.0 software. By computer molecular docking technology ，acteoside and positive inhibitors were molecularly docked with the CYP enzyme ，and the binding mode and strength of the two molecules were analyzed. RESULTS The IC 50 values of acteoside to CYP 1A2 and CYP 2A6 enzymes were more than 30 μmol/L，and those of acteoside to CYP 2D6，CYP2C19，CYP2C9 and CYP 3A4 enzymes were 24.87，21.52，12.56 and 7.55 μmol/L，respectively. The hydrogen bond and hydrophobic force could form between acteoside and CYP 3A4 enzyme，and the hydrogen bond and electrostatic interaction could form between ketoconazole and CYP 3A4 enzyme. The binding free energy of acteoside and ketoconazole to CYP 3A4 enzyme were － 10.2 and － 12.4 kcal/mol （1 kcal/mol＝4.19 kJ），respectively. CONCLUSIONS Acteoside shows moderate inhibitory effect on CYP 3A4 enzyme in liver microsomes of rats ，and its affinity is equivalent to that of positive inhibitor ；the compound shows weak inhibitory effect on other 5 CYP enzymes.

Radiofrequency ablation (RFA) is a thermal ablation technique widely used for the management of benign thyroid nodules. To date, five academic societies in various countries have reported clinical practice guidelines, opinion statements, or recommendations regarding the use of thyroid RFA. However, despite some similarities, there are also differences among the guidelines, and a consensus is required regarding safe and effective treatment in Asian countries. Therefore, a task force was organized by the guideline committee of the Asian Conference on Tumor Ablation with the goal of devising recommendations for the clinical use of thyroid RFA. The recommendations in this article are based on a comprehensive analysis of the current literature and the consensus opinion of the task force members.

Objective: To analyze the epidemiological history, clinical manifestations, treatment and the short-term prognosis of 31 cases of 2019 novel coronavirus(2019-nCoV) infection in children from six provinces (autonomous region) in northern China. Methods: A retrospective analysis of the epidemiological history, clinical symptoms, signs, laboratory examinations, chest imaging, treatment and the short-term prognosis of 31 cases of 2019-nCoV was conducted. The patients were diagnosed between January 25th, 2020 and February 21st, 2020 in 21 hospitals in 17 cities of six provinces(autonomous region) of Shaanxi, Gansu, Ningxia, Hebei, Henan and Shandong. Results: The age of the 31 children with 2019-nCoV infection was 7 years and 1 month (6 months -17 years). Nine cases (29%) were imported cases. Other 21 cases (68%) had contact with confirmed infected adults. One case (3%) had contact with asymptomatic returnees from Wuhan. Among the 31 children, 28 patients (90%) were family cluster cases. The clinical types were asymptomatic type in 4 cases (13%), mild type in 13 cases (42%), and common type in 14 cases (45%). No severe or critical type existed. The most common symptom was fever (n=20, 65%), including 1 case of high fever, 9 cases of moderate fever, 10 cases of low fever. Fever lasted from 1 day to 9 days. The fever of fifteen cases lasted for ≤3 d, while in other 5 cases lasted > 3 d. Other symptoms included cough (n=14, 45%), fatigue (n=3, 10%) and diarrhea (n=3, 9%). Pharyngalgia, runny nose, dizziness, headache and vomiting were rare. In the early stage, the total leukocytes count in peripheral blood decreased in 2 cases (6%), the lymphocytes count decreased in 2 cases (6%), and the platelet count increased in 2 cases (6%).Elevation of C-reactive protein (10%, 3/30), erythrocyte sedimentation rate(19%,4/21), procalcitonin(4%,1/28), liver enzyme(22%, 6/27) and muscle enzyme (15%, 4/27) occurred in different proportions. Renal function and blood glucose were normal. There were abnormal chest CT changes in 14 cases, including 9 cases with patchy ground glass opacities and nodules, mostly located in the lower lobe of both lungs near the pleural area. After receiving supportive treatment, the viral nucleic acid turned negative in 25 cases within 7-23 days. Among them, 24 children (77%) recovered and were discharged from hospital. No death occurred. Conclusions In this case series, 2019-nCoV infections in children from six provinces (autonomous region) in northern China are mainly caused by close family contact. Clinical types are asymptomatic, mild and common types. Clinical manifestations and laboratory examination results are nonspecific. Close contact history of epidemiology, nucleic acid detection and chest imaging are important bases for diagnosis. After general treatment, the short-term prognosis is good.

Objective To study the clinical characteristics of neonatal macrophage activation syndrome (MAS) associated with maternal rheumatic diseases and improve the understanding of neonatal MAS.Method Clinical data of MAS in a newborn infant with adult-onset Still's disease (AOSD) mother was retrospectively studied.From the establishment day of databases (CNKI,VIP,Wanfang,Pubmed and Embase) to December 2017,literature were retrieved with key words including "newborn","macrophage activation syndrome" and "hemophagocytic lymphohistiocytosis (HLH) ".Clinical features of infant MAS/HLH with maternal rheumatic diseases were summarized.Result A 27-day-old boy with AOSD mother manifested with fever,watery stools,irritability,prominent enlargement of right parotid gland and right cervical lymphadenitis.The infant was diagnosed with MAS due to coagulopathy,multiple organ dysfunction,hypofibrinogenemia and increased levels of ferritin.Anti-SSA/Ro52kD and stool rotavims antigen were positive.The infant recovered with intravenous immunoglobulin and steroids therapy.Follow-up at 2-year-old were normal.A total of 3 other cases of neonatal MAS/HLH were retrieved.All patients had high fever,hepatosplenomegaly and multiple organ dysfunction,impaired digestive system (abdominal distention,diarrhea and ascites),disseminated intravascular coagulation (2 cases),mental disorders (1 case),complete atrioventricular block (1 case) and severe hypotension (1 case).Laboratory results showed thrombocytopenia,elevated level of hepatic enzyme and serum ferritin in all patients.Targeted panel-based next generation sequencing were all negative for pathogenic gene mutations.After treatments of steroids,intravenous immunoglobulin and chemotherapy,all patients improved and ultimately cured.Conclusion In view of the impacts of the maternal rheumatic diseases on fetus,newborns with early onset high fever and hepatosplenomegaly should be suspected of MAS.Early diagnosis and effective treatment are crucial for clinical improvement.

Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected (V⁺) and uninfected (V⁻) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in V⁺ compared with V⁻ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in V⁺ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in V⁺ and V⁻ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.
Glucose-6-Phosphate Isomerase ; Glycogen Phosphorylase ; Heat-Shock Proteins ; Host-Parasite Interactions ; Malate Dehydrogenase ; Metabolism ; Polymerase Chain Reaction ; Proteome ; Reticuloendotheliosis virus ; Ribosomal Proteins ; RNA, Double-Stranded ; RNA, Messenger ; Trichomonas vaginalis* ; Trichomonas* ; Triose-Phosphate Isomerase ; Virulence

Objective To observe the efficacy of Tongmai Acupoint Plaster combined with Alprostadil injection for treatment of lower extremity arteriosclerosis obliterans (ASO). Methods Totally 180 cases of ASO Ⅱ(intermittent claudication period) patients were randomly divided into acupoint sticking group, combination group, Western medicine group, with 60 cases in each group. All groups quit smoking, had diet control, proper exercise, control of blood sugar, stop hypolipidemic and anti-coagulation medicine. The acupoint sticking group was treated with Tongmai Acupoint Plaster, each selected with the efficacy of treatment of lower limbs paralysis acupoint 10-12, alternating dressing; dressing was changed 1 time a day, 40 day as a treatment course. Western medicine group was given Alprostadil injection 10 μg, with 10 mL of normal saline, intravenous infusion, once a day, 15 d as a treatment course, for two courses, with 10 d as interval. Combination group was treated with Tongmai Acupoint Plaster combined with Alprostadil injection, the same as the application method above. The clinical efficacy, the scores of symptoms and signs, the ankle brachial index, blood lipid, the peak value of blood flow in the tibial anterior,posterior tibial, and dorsalis pedis arteries were observed and compared between the 2 groups. Results The total effective rate was 78.33% (47/60) in the acupoint sticking group, 80.00% (48/60) in the Western medicine group, and 93.33%(56/60) in the combination group, with the combination group better than the acupoint sticking group and Western medicine group (P<0.05). There was no statistical significance between acupoint sticking group and Western medicine group (P>0.05). There was statistical significance in scores of symptoms and signs, ankle brachial index before and after treatment in the three groups (P<0.05, P<0.01); There was no statistical significance after treatment (P>0.05). There was statistical significance in blood lipids before and after treatment in acupoint sticking group and combination group (P<0.05). There was statistical significance in tibial anterior, posterior tibial, and dorsalis pedis artery peak before and after treatment in the three groups (P<0.05, P<0.01). Conclusion Tongmai Acupoint Plaster has good efficacy in the treatment of ASO, equivalent with the efficacy of Alprostadil injection, which can regulate blood lipids and improve arterial blood flow. Ther combination of them has better efficacy.

The application of placebo acupuncture in randomized controlled trials in the past 10 years (2004-2014) in foreign countries was systematically reviewed to summarize the design of placebo acupuncture; according to category of diseases, advantages and disadvantages were analyzed to explore an ideal placebo acupuncture set. By retrieval in PubMed, EMBASE and OVID databases, the clinical study literature which met the inclusion criteria was searched, and the category of diseases, design of placebo acupuncture and clinical efficacy were analyzed. Totally 29 articles were included. The pain was the leading disease in category of diseases; the most commonly used application of placebo acupuncture design was telescopiform sliding blunt needle, accounting for 45. 16%; the treatment locations were non-acupoints mostly; in the needling depth, approximately 60. 00% selected non-penetrating needling and 26. 67% selected superficial needling; 28 articles indicated that acupuncture and placebo acupuncture had clinical effects, accounting for 96. 55%; 37. 93% of articles indicated acupuncture was superior to placebo acupuncture and 37. 93% of articles indicated acupuncture was not superior to placebo acupuncture. It is concluded that the design of placebo acupuncture should consider multiple factors, including main symptoms, operability, security, blinding and specific effects, and the objective indices should be selected for outcome evaluation.
Acupuncture Therapy ; instrumentation ; methods ; standards ; Humans ; Internationality ; Needles ; Placebo Effect ; Randomized Controlled Trials as Topic ; standards

Objective To explore the influence of early rapidly increased amino acid dosaging on the nitrogen balance and growth in preterm infants.Methods A total of 37 prematures admitted to our NICU within 24 hours after birth and with birth weight of 1 000 g to 1 800 g from November 2011 to January 2013 were randomly divided into study group (n =18) and control group (n =19).Parenteral nutrition (PE) was provided in both groups within 24 hours after birth.The starting amino acid dose was 1.5g/kg per day,then rose by 1 g/kg per day in study group and by 0.5 g/kg per day in control group,until to maximum of 3.5 g/kg per day.The dosage of lipids,glucose,and electrolytes in parenteral nutrition was provided routinely.The trial was double-blinded.Results In the first 5 days of PE,the nitrogen balance had no significant difference between these two groups (F =0.025,P =0.652) ; however,the time of regaining birth weight was significantly shorter in study group [(7.23±2.48) d] than in control group [(9.42±2.03) d] (t=-2.090,P=0.044).There were no differences between these two groups in trems of completely feeding time,weight gain rate,head circumference growth rate,length of hospital stay,and hospital costs (all P ＞ 0.05).Meanwhile,the rates of neonatal necrotizing enterocolitis,sepsis,mechanical ventilation administration,and damage of hepatic and renal function were also not significantly different.The administration of different dosages of amino acids did not affect the energy intake in both groups (F =1.893,P =0.218).Conclusions Prematures need early amino acid administration,with a starting dosage of more than 1.5 g/(kg · d).Rapid increase of amino acid dosing will not affect early nitrogen balance in premature infants; meanwhile,it contributes to weight gain without increasing the complications and hospitalization costs.

Marine Actinobacteria are emerging as new resources for bioactive natural products with promise in novel drug discovery. In recent years, the richness and diversity of marine Actinobacteria from the South China Sea and their ability in producing bioactive products have been investigated. The objective of this work is to isolate and identify bioactive secondary metabolites from a marine actinobacterium SCSIO 1934 derived from sediments of South China Sea. The strain was identified as a Streptomyces spieces by analyzing its 16S rDNA sequence. Streptomyces sp. SCSIO 1934 was fermented under optimized conditions and seven bioactive secondary metabolites were isolated and purified by chromatographic methods including colum chromatography over silica gel and Sephadex LH-20. Their structures were elucidated as 17-O-demethylgeldanamycin (1), lebstatin (2), 17-O-demethyllebstatin (3), nigericin (4), nigericin sodium salt (5), abierixin (6), respectively, by detailed NMR spectroscopic data (1H, 13C, COSY, HSQC and HMBC). This work provided a new marine actinobacterium Streptomyces sp. SCSIO 1934, capable of producing diverse bioactive natural products.
Anti-Bacterial Agents ; chemistry ; China ; DNA, Ribosomal ; chemistry ; genetics ; Geologic Sediments ; microbiology ; Oceans and Seas ; RNA, Ribosomal, 16S ; genetics ; Streptomyces ; chemistry ; classification ; genetics ; isolation & purification