Comparing genomes of Fructus Amomi-producing species reveals genetic basis of volatile terpenoid divergence.

Wurfbainia longiligularis and Wurfbainia villosa are both rich in volatile terpenoids and are 2 primary plant sources of Fructus Amomi used for curing gastrointestinal diseases. Metabolomic profiling has demonstrated that bornyl diphosphate (BPP)-related terpenoids are more abundant in the W. villosa seeds and have a wider tissue distribution in W. longiligularis. To explore the genetic mechanisms underlying the volatile terpenoid divergence, a high-quality chromosome-level genome of W. longiligularis (2.29 Gb, contig N50 of 80.39 Mb) was assembled. Functional characterization of 17 terpene synthases (WlTPSs) revealed that WlBPPS, along with WlTPS 24/26/28 with bornyl diphosphate synthase (BPPS) activity, contributes to the wider tissue distribution of BPP-related terpenoids in W. longiligularis compared to W. villosa. Furthermore, transgenic Nicotiana tabacum showed that the GCN4-motif element positively regulates seed expression of WvBPPS and thus promotes the enrichment of BPP-related terpenoids in W. villosa seeds. Systematic identification and analysis of candidate TPS in 29 monocot plants from 16 families indicated that substantial expansion of TPS-a and TPS-b subfamily genes in Zingiberaceae may have driven increased diversity and production of volatile terpenoids. Evolutionary analysis and functional identification of BPPS genes showed that BPP-related terpenoids may be distributed only in the Zingiberaceae of monocot plants. This research provides valuable genomic resources for breeding and improving Fructus Amomi with medicinal and edible value and sheds light on the evolution of terpenoid biosynthesis in Zingiberaceae.

Ultra-low-dose rituximab is effective for the treatment of patients with minimal change disease - A retrospective study.

BACKGROUND PURPOSE: Rituximab (RTX),an anti-CD20 monoclonal antibody can effectively treat minimal change disease (MCD),with low toxicity and a reduced steroid dosage. The optimal dosage of RTX for treating MCD remains unclear. This study aimed to investigate the efficacy of an ultra-low-dose regimen of RTX (100 mg per week for 4 weeks) for treating MCD. METHODS: We retrospectively analyzed clinical data from 31 patients with MCD who received RTX. Seventeen patients received ultra-low-dose RTX (ULD-RTX) therapy, and 14 patients received standard-dose RTX (SD-RTX) therapy (500 mg weekly for 4 weeks). All patients were followed up for at least 6 months. RESULTS: Both groups showed significant increases in the serum albumin levels and notable decreases in the urinary protein levels in the 1st and 6th months after RTX therapy. There were no significant differences in the plasma albumin or urinary protein levels between the groups (p > 0.05). B-cell depletion was observed in all patients after 1 month of RTX administration. At 6 months after RTX treatment, the remission rate was 93% in the SD-RTX group and 88% in the ULD-RTX group (p > 0.05). The ULD-RTX therapy incurred lower costs than did the SD-RTX therapy. One patient in the SD-RTX group developed community-acquired pneumonia. CONCLUSION: Ultra-low-dose RTX is effective at inducing remission in patients with MCD at a lower cost.

Origin of the negative longitudinal piezoelectric effect and electric auxetic effect in hexagonal AIBIVCV semiconductors.

Hexagonal ABC semiconductors with a polar structure are potential candidates for piezoelectric applications. The intriguing negative longitudinal piezoelectric effect (NLPE) and electric auxetic effect (EAE) may exist in these materials, and establishing the structure-property relation provides physical insights into the underlying mechanisms responsible for these phenomena. In this work, using first-principles calculations, we investigate the piezoelectric response in a class of hexagonal AIBIVCV (A = Li, Na, and K; B = Ge and Sn; C = N, P, As, and Sb) semiconductors. We demonstrate that the quasi-layered structure with contrasting interlayer and intralayer bonding strengths plays a crucial role in the longitudinal piezoelectric response. In this class of materials, we identify 11 compounds out of the 24 candidates possessing the NLPE. We find that the NLPE tends to occur when the quasi-layered structure is pronounced. Moreover, we identify an unusual coexistence of negative longitudinal and transverse piezoelectric responses, and hence the compounds possessing the NLPE are electric auxetic materials as well. This work provides a simple guide for the search of piezoelectrics with desired responses.

[Comparison of catalytic functions and expression patterns of two pinene synthases from Wurfbainia villosa].

Wurfbainia villosa fruit is rich in volatile terpenoids, among which pinene is one of the main components and has anti-inflammatory, antibacterial, anti-tumor, and other pharmacological activities. This research group found that W. villosa fruits were rich in α-pinene by GC-MS, and terpene synthase(WvTPS63, formerly known as AvTPS1) with ß-pinene as the main product was cloned and identified, but α-pinene synthase had not been identified. In this study, based on the genome data of W. villosa, we screened and found WvTPS66 with highly similar sequences to WvTPS63, identified enzyme functions of WvTPS66 in vitro, and performed a comparative analysis of sequence, catalytic function, expression pattern, and promoter with WvTPS63. Multiple sequence alignment showed that the amino acid sequences of WvTPS63 and WvTPS66 were highly similar and the conservative motif of terpene synthase was almost identical. In vitro enzymatic experiments on catalytic functions showed that both could produce pinene, and the main product of WvTPS63 was ß-pinene, while that of WvTPS66 was α-pinene. Expression pattern analysis showed that WvTS63 was highly expressed in flowers, WvTPS66 was expressed in the whole plant, and the highest expression level was found in the pericarp, which indicated that it might be mainly responsible for the synthesis of α-pinene in fruits. In addition, promoter analysis revealed the presence of multiple regulatory elements related to stress response in the promoter regions of both genes. The findings of this study can provide a reference for the functional study of terpene synthase genes and new genetic elements for pinene biosynthesis.

Development of a Fully Continuous-Flow Approach Towards Asymmetric Total Synthesis of Tetrahydroprotoberberine Natural Alkaloids.

Continuous flow synthetic technologies had been widely applied in the total synthesis in the past few decades. Fully continuous flow synthesis is still extremely focused on multi-step synthesis of complex natural pharmaceutical molecules. Thus, the development of fully continuous flow total synthesis of natural products is in demand but challenging. Herein, we demonstrated the first fully continuous flow approach towards asymmetric total synthesis of natural tetrahydroprotoberberine alkaloids, (-)-isocanadine, (-)-tetrahydropseudocoptisine, (-)-stylopine and (-)-nandinine. This method features a concise linear sequence involving four chemical transformations and three on-line work-up processing in an integrated flow platform, without any intermediate purification. The overall yield and enantioselectivity of this four-step continuous flow chemistry were up to 50 % and 92 %ee, respectively, in a total residence time of 32.5 min, corresponding to a throughput of 145 mg/h.

Conductive polymer sorbent for extraction and determination of resveratrol and polydatin in Polygonum cuspidatum root samples.

INTRODUCTION: The quantitative analysis of trace resveratrol and polydatin in plant tissues is suitable for elucidation of the compounds' mechanisms of action. OBJECTIVES: The main objective of this work was to develop a feasible and effective sample pretreatment method to measure the concentrations of resveratrol and polydatin in complex samples. METHODOLOGY: A polymer sorbent, poly(2-mercaptobenzimidazole), was electrochemically prepared and utilized for selective extraction, while resveratrol and polydatin were used as target analytes. The sorbent was characterized by cyclic voltammetry, scanning electron microscopy and Fourier transform infrared spectroscopy. After extraction and elution, the analytes were analyzed by a Thermo U3000 HPLC system. Several affecting parameters, including the volume of elution solution, sample pH value, sample flow rate and sample volume, were evaluated and optimized. RESULTS: The proposed method showed good linearity with low limits of detection (from 0.5 to 0.8 ng·mL-1 ) and ideal accuracy with spiked recoveries from 81.30% to 99.16%. A good enrichment factor (more than 200-fold) together with good sensitivity was obtained with this method. Analysis of resveratrol and polydatin in Polygonum cuspidatum samples by this method is efficient. CONCLUSION: The method developed in this work exhibits several significant merits, including easy operation and high extraction efficiency, indicating that electrochemically prepared polymer sorbent is useful for sample pretreatment and analysis of traditional Chinese medicine samples.

Second victim experience and support desire among nurses working at regional levels in China.

AIM: The aim of this study was to describe and analyse the degree of second victim syndrome and the desire for supports among nurses working in regional hospitals in China. BACKGROUND: The evidence on the prevalence of second victim among health care workers remains inconsistent and has rarely focused on the regional level. METHODS: A quantitative, descriptive, survey-based, online, cross-sectional study was conducted among 1,194 nurses in three regional hospitals. RESULTS: A total of 918 (76.88%) nurses, who reported have had experienced patient safety incidents, were selected for the final analysis. The mean score for the Chinese version of the Second Victim Experience and Support Tool (C-SVEST) was (65.58 ± 10.05). Psychological distress (15.91 ± 2.99) and practice distress (15.26 ± 4.32) had the highest score. The mean score for the desired form of support was (4.29 ± 0.614). The option 'the opportunity to get guidance and suggestions for future work' was rated the most desired. CONCLUSION: Nurses working at the regional level reported a similar degree of second victim experience and support desire, while the prevalence was much higher. IMPLICATIONS FOR NURSING MANAGEMENT: The second victim phenomenon has become increasingly complex and challenging and deserves more attention. Not only Safety-I but also Safety-II approaches are suggested to integrated to patient safety.

Impact of intrauterine hypoxia on adolescent and adult cognitive function in rat offspring: sexual differences and the effects of spermidine intervention.

Intrauterine hypoxia (IUH) affects the growth and development of offspring. It remains unclear that how long the impact of IUH on cognitive function lasts and whether sexual differences exist. Spermidine (SPD) has shown to improve cognition, but its effect on the cognitive function of IUH offspring remains unknown. In the present study we investigated the influence of IUH on body weight and neurological, motor and cognitive function and the expression of APP, BACE1 and Tau5 proteins in brain tissues in 2- and 4-month-old IUH rat offspring, as well as the effects of SPD intervention on these parameters. IUH rat model was established by treating pregnant rats with intermittent hypoxia on gestational days 15-21, meanwhile pregnant rats were administered SPD (5 mg·kg-1·d-1;ip) for 7 days. Neurological deficits were assessed in the Longa scoring test; motor and cognitive functions were evaluated in coat hanger test and active avoidance test, respectively. We found that IUH decreased the body weight of rats in both sexes but merely impaired motor and cognitive function in female rats without changing neurological function in the rat offspring of either sex at 2 months of age. For 4-month-old offspring, IUH decreased body weight in males and impaired neurological function and increased cognitive function in both sexes. IUH did not affect APP, BACE1 or Tau5 protein expression in either the hippocampus or cortex of all offspring; however, it increased the cortical Tau5 level in 2-month-old female offspring. Surprisingly, SPD intervention prevented weight loss. SPD intervention reversed the motor and cognitive decline caused by IUH in 2-month-old female rat offspring. Taken together, IUH-induced cognitive decline in rat offspring is sex-dependent during puberty and can be recovered in adult rats. SPD intervention improves IUH-induced cognitive and neural function decline.

Boronate affinity solid-phase extraction of cis-diol compounds by a one-step electrochemically synthesized selective polymer sorbent.

A rational-designed conductive sorbent, poly(thiophene-3-boronic acid) electrochemically deposited on a carbon fiber bundle, was applied for boronate affinity extraction. The coated carbon fiber bundle packed into a poly(ether ether ketone) tube was then successfully used for online solid-phase microextraction-high-performance liquid chromatography analysis of cis-diol compounds. Three kinds of catecholamines (namely, adrenaline, noradrenaline, and dopamine) were used as the test analytes. Good extraction efficiency (more than 600-fold), low limits of detection (0.2 ng·mL-1 for adrenaline, 0.1 ng·mL-1 for noradrenaline and dopamine), and wide linear ranges were obtained. The method was demonstrated to be efficient for analysis of catecholamines in spiked plasma samples, with good recoveries in the range of 92.5-95.71%. This work exhibited several significant advantages, including ease of use, high specificity, and high extraction efficiency. It has been demonstrated that such an electrochemically synthesized sorbent has great potential for research involving cis-diol compounds. Graphical abstract Electrochemical modification, packing of fiber bundles, and extraction mechanism. PEEK poly(ether ether ketone).

Polymeric monolith column composited with multiwalled carbon nanotubes-ß-cyclodextrin for the selective extraction of psoralen and isopsoralen.

A polymeric column that contains multiwalled carbon nanotubes-ß-cyclodextrin composite was developed. The composite was wrapped into the poly(butyl methacrylate-ethylene dimethacrylate) monolith column (0.76 mm id and 10 cm in length). The column was then applied for the online solid-phase microextraction of psoralen and isopsoralen from Fructus Psoraleae. Following microextraction, the coumarins were quantified by high-performance liquid chromatography with C18 separation column and UV detection. The effects of sample flow rate, sample volume, and pH value were optimized. The method showed low limits of detection (20 pg/mL, S/N = 3) for both psoralen and isopsoralen. Finally the method was successfully applied to the determination of psoralen and isopsoralen in spiked herb extracts and rat plasma where it gave recoveries that ranged between 93.2 and 102.1%. The empty hydrophobic cavities of ß-cyclodextrin and the hydrophobicity of multiwalled carbon nanotubes provided specific extraction capability for psoralen and isopsoralen.

Novel polymeric monolith materials with a ß-cyclodextrin-graphene composite for the highly selective extraction of methyl jasmonate.

A novel polymeric monolith column with a  ß-cyclodextrin-graphene composite was prepared for extraction of methyl jasmonate. A simple, sensitive, and effective polymeric monolith microextraction with high-performance liquid chromatography method has been presented for the determination. To carry out the best microextraction efficiency, several parameters such as sample flow rate, sample volume, and sample pH value were systematically optimized. In addition, the method validation showed a wide linear range of 5-2000 ng/mL, with a good linearity and low limits of detection for methyl jasmonate. The proposed method was successfully applied for the determination of methyl jasmonate in wintersweet flowers with recoveries of 90.67%. The result was confirmed by high-performance liquid chromatography with mass spectrometry.

[The Antimicrobial Resistance Analysis of 73 Strains of Streptococcus pneumoniae Isolated from Infant Respiratory Tract].

OBJECTIVE: To preliminary study of the resistance mechanisms of S. pneumoniae (S. pn) by determining the resistance rates and gene of S. pn isolated from the lower respiratory tract infection infants. METHODS: Drug susceptibility test with disk diffusion and broth micro-dilution was conducted to evaluate the resistance rates of 73 strains of S. pn isolated from the lower respiratory tract infection infants to penicillin, levofloxacin and other 10 antibiotics. PCR method was used to analysis the antimicrobial resistant genes tet M, mef A, erm A, erm B and int Tn of the isolates. RESULTS: The antibiotic resistance rates of the S. pn isolates to erythromycin, clindamycin and tetracycline were 95. 9%, 94. 5%, 87. 7% and 0% to vancomycin when tested with disk diffusion method. The antibiotic resistance rates of these isolates to penicillin, cefotaxime and ceftriaxone were 45. 2%, 47. 9% and 46. 6% respectively when tested with broth micro-dilution method. The carrier frequencies of tet M, mef A, erm A, erm B, int Tn genes in the 73 isolates were 91. 8%, 63. 0%, 58. 9%, 39. 7% and 61. 6% respectively. CONCLUSION: The S. pn strains isolated from infant respiratory tract in Chengdu perform a serious drug resistance problem, especially to routine antibiotics like erythromycin, clindamycin and tetracycline and cephalosporin, the resistance rate to levofloxacin, chloramphenicol remained at a low level; the resistance to tetracycline was closely related with the tet M gene fragment, the resistance to macrolide was mainly decided by active efflux pump and secondarily by the alternation of gene targeting, int Tn had close relation with tet M, erm B.

BeeSpace Navigator: exploratory analysis of gene function using semantic indexing of biological literature.

With the rapid decrease in cost of genome sequencing, the classification of gene function is becoming a primary problem. Such classification has been performed by human curators who read biological literature to extract evidence. BeeSpace Navigator is a prototype software for exploratory analysis of gene function using biological literature. The software supports an automatic analogue of the curator process to extract functions, with a simple interface intended for all biologists. Since extraction is done on selected collections that are semantically indexed into conceptual spaces, the curation can be task specific. Biological literature containing references to gene lists from expression experiments can be analyzed to extract concepts that are computational equivalents of a classification such as Gene Ontology, yielding discriminating concepts that differentiate gene mentions from other mentions. The functions of individual genes can be summarized from sentences in biological literature, to produce results resembling a model organism database entry that is automatically computed. Statistical frequency analysis based on literature phrase extraction generates offline semantic indexes to support these gene function services. The website with BeeSpace Navigator is free and open to all; there is no login requirement at www.beespace.illinois.edu for version 4. Materials from the 2010 BeeSpace Software Training Workshop are available at www.beespace.illinois.edu/bstwmaterials.php.

α-Lipoic acid chemistry: the past 70 years.

α-Lipoic acid (ALA) is a naturally occurring sulfur-containing fatty acid with high antioxidant activity. It is also used to treat diabetes, nerve pain, weight loss, heart disease, and primary mitochondrial disorders. Moreover, numerous therapeutic agents have been studied for managing other clinical conditions, including for anticancer, anti-HIV, anti-inflammatory, and anti-AD treatments. The medicinal importance of ALA, especially its biologically active form (R-ALA), has attracted considerable attention from synthetic chemists in industrial and academic fields. In this review, we discuss synthetic approaches to ALA and R-ALA over the past 70 years (1952 to the present), which will help medicinal chemists further develop novel routes for their synthesis.

Development of novel S-N3-DABO derivatives as potent non-nucleoside reverse transcriptase inhibitors with improved potency and selectivity.

Following on our initial discovery of S-CN-DABOs as non-nucleoside reverse transcriptase inhibitors (NNRTIs), a series of novel S-N3-DABO derivatives F1-F31 were developed by substituting the cyano group of S-CN-DABOs with azide group. Some of these compounds were conferred significantly increased potency against wild-type HIV-1 and clinically observed mutant strains. Remarkably, the best compound F10 exerted a 7-fold improvement in potency (EC50 = 0.053 µM) and 12.5-fold higher selectivity (SI = 6818) in MT-4 cells infected with wild-type HIV-1, compared to that of the parent compound B1 (EC50 = 370 nM, SI = 547). The anti-HIV-1 activity of F10 against the tested mutant strains was prominently enhanced. For wild-type reverse transcriptase, it was approximately 19-fold more potent (IC50 = 0.080 µM) than B1 (IC50 = 1.51 µM). It was not found that this analog had significant inhibition of hERG, CYP, and acute toxicity after a single dose of F10 (1.0 g/kg).

Collagen VI-related myopathies: clinical variability, phenotype-genotype correlation and exploratory transcriptome study.

Collagen VI-related myopathies are a group of disorders that cause muscle weakness and joint contractures with significant variability in disease severity among patients. Here we report the clinical and genetic characteristics of 13 Chinese patients. Detailed histological, radiological and muscle transcriptomic evaluations were also conducted for selected representative patients. Across the cohort, fifteen putative disease causal variants were identified in three genes encoding collagen VI subunits, COL6A1 (n=6), COL6A2 (n=5), and COL6A3 (n=4). Most of these variants (12/15, 80%) were dominant negative and occurred at the triple helical domain. The rest (3/15, 20%) were located at the C-terminus. Two previously unreported variants, an in-frame mutation (COL6A1:c.1084_1092del) and a missense mutation (COL6A2:c.811G>C), were also noted. The transcriptome data from the muscle biopsies of two patients in the study with dominant negative mutations [COL6A2:c.811G>C and COL6A1:c.930+189C>T] supports the accepted aetiology of Collagen VI myopathy as dysfunction of the extracellular matrix. It also suggests there are perturbations to skeletal muscle differentiation and skeletal system development. It should be noted that although the phenotypes of patients can be mostly explained by the position and dominant-negative effect of the variants, exceptions and variability still exist and have to be reckoned with. This study provides valuable data explaining the varying severity of phenotypes among ethnically Chinese patients.

Acute kidney injury as the first sign of spontaneous renal vein thrombosis: report of 2 cases.

Spontaneous renal vein thrombosis (RVT) is very rare in the absence of nephrotic syndrome. It is more common in newborns and infants. RVT should always be included in the differential diagnosis of flank pain and hematuria, and because RVT can induce acute renal injury. A 19-year-old man was admitted to our hospital because he complained of right flank pain and oliguria for 3 days. Another patient, a 24-year-old man, complained of a severe and sudden onset of bilateral flank pain and anuria for a day. They were both healthy before they developed the described symptoms and had different levels of decrease in renal function when they visited the hospital. Color Doppler ultrasonography revealed RVT in both the patients. The patients received therapy, including anticoagulation and thrombolysis, following their diagnoses, and they recovered in a few days.

BSQA: integrated text mining using entity relation semantics extracted from biological literature of insects.

Text mining is one promising way of extracting information automatically from the vast biological literature. To maximize its potential, the knowledge encoded in the text should be translated to some semantic representation such as entities and relations, which could be analyzed by machines. But large-scale practical systems for this purpose are rare. We present BeeSpace question/answering (BSQA) system that performs integrated text mining for insect biology, covering diverse aspects from molecular interactions of genes to insect behavior. BSQA recognizes a number of entities and relations in Medline documents about the model insect, Drosophila melanogaster. For any text query, BSQA exploits entity annotation of retrieved documents to identify important concepts in different categories. By utilizing the extracted relations, BSQA is also able to answer many biologically motivated questions, from simple ones such as, which anatomical part is a gene expressed in, to more complex ones involving multiple types of relations. BSQA is freely available at http://www.beespace.uiuc.edu/QuestionAnswer.

Expansion of the S-CN-DABO scaffold to exploit the impact on inhibitory activities against the non-nucleoside HIV-1 reverse transcriptase.

The α-cyanoarylmethyl-3, 4-dihydropyrimidin-4(3H)-ones (S-CN-DABOs) were reported as a kind of reverse transcriptase inhibitors of human immunodeficiency virus type-1 (HIV-1) by our group in 2007. In this paper, we proposed to expand the S-CN-DABO scaffold to enrich the structure-activity relationship (SAR) of the phenyl ring that was predicted to be located in the W229 hydrophobic pocket. Thirty-nine S-CN-DABO derivatives were manufactured to explore the impact on inhibitory activities against the non-nucleoside HIV-1 reverse transcriptase. These analogues displayed up to low nanomolar activity against wild-type (WT) HIV-1 and good activity against several clinically relevant resistant mutant viruses, especially rilpivirine-associated resistant mutant E138K strain. The inhibitory ability toward the RT enzyme was significantly improved. Compound B23 with a 2, 6-difluoro-phenyl group showed inhibitory effects with an EC50 value of 20.8 nM against HIV-1 WT strain, and an EC50 of 50 nM targeting mutant E138K, which were about 20-fold better than the lead compound B1. Molecular docking analysis elucidated the biological activity and offered a structural insight for follow-up research. In addition, compound B23 also showed favorable drug-like properties in vitro and in vivo. There was no significant inhibition of hERG (IC50 > 40 µM), no apparent CYP enzymatic inhibitory activity and acute toxicity in mouse models. Perfect oral bioavailability of compound B23 was revealed (F = 164%, SD rats). In summary, these S-CN-DABOs compounds could be further optimized and modified for promising drug candidates in anti-HIV clinical therapy.

Development of a High-Titer Culture Medium for the Production of Cholesterol by Engineered Saccharomyces cerevisiae and Its Fed-Batch Cultivation Strategy.

Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.