Quality Evaluation of Shexiang Tongxin Dropping Pill Based on HPLC Fingerprints Combined with HPLC-Q-TOF-MS/MS Method.

Shexiang Tongxin Dropping Pill (STP) is a composite formula of traditional Chinese medicine that is widely used for the treatment of cardiovascular diseases. It consists of seven medicinal extracts thereof or materials, including Bufonis venenum, synthetic Moschus, Panax ginseng, Bovis calculus artifactus, Bear bile powder, Salvia miltiorrhiza Bge and synthetic borneol. However, it is considerably difficult to evaluate the quality of STP due to its complex chemical compositions. This paper was designed to explore a comprehensive and systematic method combining fingerprints and chemical identification for quality assessment of STP samples. Twenty batches of STP samples were analyzed by high-performance liquid chromatography (HPLC) and high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry. Ten common peaks were detected by HPLC fingerprint similarity evaluation system. Meanwhile, 100 compounds belonging to 4 structural characteristics, including 23 bufadienolides, 36 organic acids, 34 saponins and 7 other types, were systematically identified as the basic components in STP. This study could be used for clarifying the multiple bioactive substances and developing a comprehensive quality evaluation method of STP.

Acute acquired concomitant esotropia with congenital paralytic strabismus: A case report.

BACKGROUND: An unusual case of acute acquired concomitant esotropia (AACE) with congenital paralytic strabismus in the right eye is reported. CASE SUMMARY: A 23-year-old woman presented with complaints of binocular diplopia and esotropia of the right eye lasting 4 years and head tilt to the left since 1 year after birth. The Bielschowsky head tilt test showed right hypertropia on a right head tilt. She did not report any other intracranial pathology. A diagnosis of AACE and right congenital paralytic strabismus was made. Then, she underwent medial rectus muscle recession and lateral rectus muscle resection combined with inferior oblique muscle myectomy in the right eye. One day after surgery, the patient reported that she had no diplopia at either distance or near fixation and was found to be orthophoric in the primary position; furthermore, her head posture immediately and markedly improved. CONCLUSION: In future clinical work, in cases of AACE combined with other types of strabismus, we can perform conventional single surgery for both at the same time, and the two types of strabismus can be solved simultaneously.

Redescription and Molecular Characterization of Trichotylenchus dispersus (Nematoda: Dolichodoridae) in China.

A species of Trichotylenchus nematode was isolated from the rhizosphere of banana root in Leizhou City, Guangdong Province, China. The species assumes the following characteristics: open C-shaped body; head offset from body; lateral field with three incisures, pharyngeal and tail regions irregularly areolated; stylet 18.6-20.7 µm long; pharyngeal gland not extending over intestine; fibrous tissue present in the intestine; post-anal intestinal sac present; elongate-subcylindroid tail, bluntly conoid terminus, lack of striations, and containing 34-44 annuli. In addition, scanning electron microscopy was used to elucidate some morphological details, but only some juveniles were observed. Partial 18S rRNA, ITS, and 28S D2-D3 expansion sequences were amplified with universal primers and deposited in GenBank under accession numbers ON622716, ON622717, and ON622714, respectively. Here, this species was identified as T. dispersus [(Siddiqi & Sharma, 1995) Geraert, 2011].

A novel ATPase gene, Ab-atps, plays an important role in the interaction of rice and white tip nematode, Aphelenchoides besseyi.

Plant kinases containing the LysM domain play important roles in pathogen recognition and self-defense reactions. And it could recognize microbe-associated molecules including chitin and other polypeptides. The white tip nematode Aphelenchoides besseyi is a migratory parasitic nematode that infects plant shoots. It is distributed over almost all rice-producing areas and causes up to 50% economic losses. The rice OsRLK3 gene was a defense-related LysM kinase gene of rice. This study showed that the rice LysM kinase OsRLK3 could be induced by flg22, jasmonic acid, salicylic acid, and chitin. An interaction gene, Ab-atps from A. besseyi, was identified by screening the interaction between the rice gene OsRLK3 and an A. besseyi cDNA library using yeast two-hybrid screening. Ab-atps is a novel ATP synthase gene with a full length of 1341 bp, coding for 183 amino acids. The mRNA of Ab-atps was located in the esophagus and reproductive system of A. besseyi. The expression of Ab-atps was assessed at different developmental stages of the nematode and found to be the highest in the juvenile, followed by the egg, female, and male. Reproduction was significantly decreased in nematodes treated with Ab-atps double-stranded RNA (dsRNA) (p < 0.05). Transient expression experiments showed that Ab-ATPS-GFP was distributed in the nucleus, cytoplasm, and cell membrane, and Ab-ATPS-GFP triggered plant cell death. OsRLK3 was expressed significantly higher at 0.5 day and 1 day (p < 0.05) in rice plants inoculated with nematodes treated with Ab-atps dsRNA and gfp dsRNA for 0.5-7 days, respectively. Further, OsRLK3 expression under Ab-atps dsRNA treatment was significantly lower than with gfp dsRNA treatment at 0.5 day (p < 0.05) and significantly higher than with gfp dsRNA treatment at 1 day (p < 0.05). These results suggest that rice OsRLK3 could interact with A. besseyi Ab-atps, which plays an important role in growth, reproduction, and infection of the nematode. Our findings provide a theoretical basis to further understand the parasitic strategy of A. besseyi and its interaction mechanism with host plants, suggesting new ideas and targets for controlling A. besseyi.

Development of a fluorescent probe for detecting Al3+ in cooked wheaten food based on phosphonic acid group functionalized polythiophene derivatives.

As an unnecessary trace element, the content of aluminium in biological systems should be strictly controlled. Therefore, it was necessary to develop a convenient method for detection of aluminium ions. In this study, a fluorescent probe based on polythiophene derivatives was developed and used to detect Al3+ in Chinese traditional pasta. The fluorescence of this probe showed a significant decrease in hexamethylenetetramine-HCl buffer solution (pH 5) when Al3+ was present. In addition, the probe exhibited good sensitivity and selectivity to Al3+ over other metal ions when EDTA was used as the masking agent. Fluorescence intensity had a good linear relationship with the Al3+ concentration in the range 0.1-10 µM and the limit of detection for Al3+ was 39 nM. Furthermore, the probe was successfully applied to detect Al3+ in food samples and the results were consistent with ICP-AES.

[Seasonal Characteristics and Source Analysis of Water-soluble Inorganic Ions in PM2.5 in Suqian City].

To study the seasonal pollution characteristics and sources of water-soluble inorganic ions in atmospheric PM2.5 in Suqian City, 171 samples were collected at three monitoring points, which were in the water vapor channel, from May 2017 to January 2018. The mass concentrations of PM2.5 and nine water-soluble inorganic ions were analyzed. The results showed that the annual average concentration of water-soluble inorganic ions in PM2.5 in Suqian City was (44.08±34.61) µg ·m-3, accounting for 41.8% of PM2.5. The concentrations of these species were in the order of ρ(NO3-) > ρ(SO42-) > ρ(NH4+) > ρ(ρl-) > ρ(Na+) > ρ(Ca2+) > ρ(K+) > ρ(F-) > ρ(Mg2+); NO3-, SO42-, and NH4+ accounted for 75.6% of the total water-soluble ions. The annual average ratio of ρ(NO3-) to ρ(SO42-) was 1.53±0.88, indicating that mobile sources contributed more to PM2.5 pollution. Based on the correlation analysis of NH4+ and SO42-, NO3- may exist in the form of (NH4)2 SO4, NH4HSO4, or NH4NO3. According to the principal component analysis, secondary transformation, industrial pollution, biomass burning, and dust were the major sources of water-soluble inorganic ions. PM2.5concentrations were positively related to relative humidity in winter. Water vapor transmission is more likely to promote PM2.5 accumulation in winter.

Author Correction: Evaluation of Stratiolaelaps scimitus (Acari: Laelapidae) for controlling the root-knot nematode, Meloidogyne incognita (Tylenchida: Heteroderidae).

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Evaluation of Stratiolaelaps scimitus (Acari: Laelapidae) for controlling the root-knot nematode, Meloidogyne incognita (Tylenchida: Heteroderidae).

Root-knot nematodes are one of the most harmful plant-parasitic nematodes (PPNs). In this paper, the predation of Stratiolaelaps scimitus against Meloidogyne incognita was tested in an individual arena, and the control efficiency of the mite on the nematode in the water spinach (Ipomoea aquatica) rhizosphere was studied with a pot experiment. The results showed that S. scimitus could develop normally and complete its life cycle by feeding on second-stage juveniles of M. incognita (Mi-J2). The consumption rate of a 24 h starving female mite on Mi-J2 increased with the increase of prey density at 25 °C. Among the starvation treatments, the nematode consumption rate of a female mite starved for 96 h at 25 °C was highest; and among temperature treatments, the maximum consumption rate of a 24 h starving female mite on Mi-J2 was at 28 °C. The number of M. incognita in the spinach rhizosphere could be reduced effectively by releasing S. scimitus into rhizosphere soil, and 400 mites per pot was the optimum releasing density in which the numbers of root knots and egg masses decreased by 50.9% and 62.8%, respectively. Though we have gained a greater understanding of S. scimitus as a predator of M. incognita, the biocontrol of M. incognita using S. scimitus under field conditions remains unknown and requires further study.

Expression analysis of proteasome maturation protein (POMP) gene in Liaoning Cashmere goat.

Liaoning Cashmere goat is a precious genetic resource of China. To explore the relationship between POMP and cashmere growth, we analyzed the expression of POMP. POMP encodes a hudrophilic protein which is most closely related to bos. RT-PCR showed that POMP was expressed in skin, heart, liver, spleen, lung, and kidney tissues. Real-time PCR showed that the expression of POMP was more active in the secondary hair follicles than the primary hair follicles in anagen. In situ hybridization showed that POMP was obviously expressed in the Inner Root Sheath (IRS) but no expression in Outer Root. The treatment of fibroblasts with melatonin (MT), fibroblast growth factors 5 (FGF5) and insulin-like growth factors 1 (IGF-1) showed that MT/FGF5/IGF-1 much performance for inhibiting the expression of POMP; MT + FGF5 inhibited the expression of POMP; MT + IGF-1 promoted the expression of POMP. When Noggin expression is decreased by siRNA, the expression of POMP is inhibited. To sum up, POMP strongly expressed in the root sheath of hair follicles, related to the development of the primary and secondary hair follicle; In addition, by adding MT/FGF5/IGF-1 or interfering with the Noggin expression to regulate the expression of POMP, to control the growth of Liaoning Cashmere goat cashmere.

RNA-Seq revealed that infection with white tip nematodes could downregulate rice photosynthetic genes.

White-tip nematode, Aphelenchoides besseyi is a kind of widely distributed migratory parasitic nematode that can infect plant shoots. Transcriptome sequencing of plant parasitic nematodes and their host plants is helpful for understanding their interaction relationship. This study first reported expression patterns of defense-related genes in rice, and rice transcriptomes at different periods after infection with A. besseyi. The result showed that the defense response pathways of rice changed obviously in the early stage of A. besseyi infection, including upregulated salicylic acid and jasmonate pathways and a downregulated ethylene pathway. Transcriptome analysis results suggested that A. besseyi infection was associated with the downregulation of multiple genes related to photosynthesis with possible suppression of the photosynthetic activity. It suggested that the photosynthesis system of rice could be suppressed by infections of migratory nematodes, including A. besseyi and Hirschmanniella oryzae, but was stimulated by that of a sedentary nematode, Meloidogyne graminicola, by comparing our study with the reported transcriptome. OS09G0417800 (OsWRKY62) might play an important role in the interaction of migratory nematodes and rice. It also indicated that the infection strategy of both A. besseyi and the reported migratory nematode H. oryzae was similar to that of the fungal pathogen Magnaporthe grisea. These results provided an interesting starting point to elucidate the mechanism of the interaction between rice and A. besseyi, as well as the host and migratory plant nematodes.

Identification and Characterization of a Fatty Acid- and Retinoid-Binding Protein Gene (Ar-far-1) from the Chrysanthemum Foliar Nematode, Aphelenchoides ritzemabosi.

The chrysanthemum foliar nematode (CFN), Aphelenchoides ritzemabosi, is a migratory, plant-parasitic nematode that is widely distributed and infects the aboveground parts of many plants. The fatty acid- and retinoid-binding proteins (FAR) are nematode-specific proteins that are involved in the development, reproduction, and infection of nematodes and are secreted into the tissues to disrupt the plant defense reaction. In this study, we obtained the full-length sequence of the FAR gene (Ar-far-1) from CFN, which is 727 bp and includes a 546 bp ORF that encodes 181 amino acids. Ar-FAR-1 from CFN has the highest sequence similarity to Ab-FAR-1 from A. besseyi, and they are located within the same branch of the phylogenetic tree. Fluorescence-based ligand-binding analysis confirmed that recombinant Ar-FAR-1 was bound to fatty acids and retinol. Ar-far-1 mRNA was expressed in the muscle layer, intestine, female genital system, and egg of CFN, and more highly expressed in females than in males among the four developmental stages of CFN. We demonstrated that the reproduction number and infection capacity of CFN decreased significantly when Ar-far-1 was effectively silenced by in vitro RNAi. Ar-far-1 plays an important role in the development, reproduction, infectivity, and pathogenesis of CFN and may be used as an effective target gene for the control of CFN. The results provide meaningful data about the parasitic and pathogenic genes of CFN to study the interaction mechanism between plant-parasitic nematodes and hosts.

Three new species, one new genus and subfamily of Dorylaimida (de Man, 1876) Pearse, 1942, and revisions on the families Tylencholaimellidae Jairajpuri, 1964 and Mydonomidae Thorne, 1964 (Nematoda: Dorylaimida).

Three new species of the order Dorylaimida (de Man, 1876) Pearse, 1942 were identified and described. Paratylencholaimus sanshaensis gen. nov. sp. nov. from Hainan is proposed as a new member of the family Tylencholaimellidae Jairajpuri, 1964. Paratylencholaimus gen. nov. is close to Phellonema Thorne, 1964 and Goferus Jairajpuri & Ahmad, 1992 but can be differentiated mainly by having basal part of odontophore rod-like and without knobs, and basal part of pharynx expanded gradually. Tylencholaimus zhongshanensis sp. nov. from Guangdong and Dorylaimoides shapotouensis sp. nov. from the Ningxia Hui Autonomous Region are also described herein. Phylogenetic analyses based on the 18S rDNA and the D2-D3 region of the 28S rDNA support that the three new species are valid. The classifications of the families Tylencholaimellidae and Mydonomidae Thorne, 1964 are revised mainly based on the analysis of the morphology of odontostyle and odontophore. After these revisions, Paratylencholaiminae subfam. nov. including Paratylencholaimus gen. nov. and Goferus is proposed. Athernema and Agmodorus of Tylencholaimellidae are transferred into Mydonomidae, and the subfamily Athernematinae of Tylencholaimellidae is dismissed. The main characteristics of the family Mydonomidae and Tylencholaimellidae are revised. Keys to the genera of Mydomonidae and Tylencholaimellidae are included.

Developed a novel sensor based on fluorescent graft conjugated polymer for the determination of aristolochic acid in traditional Chinese medicine.

A novel fluorescent graft conjugated polymer (poly (2, 5-bis (Polyethylene glycol oxybutyrate)-1, 4-phenylethynylene-alt-1, 4-phenyleneethynylene, PPE-OB-PEG) has been designed and synthesized for the determination of aristolochic acid (AA). The detection conditions and detection characters of PPE-OB-PEG were systematically explored in this work. The fluorescence intensity of PPE-OB-PEG changes with the different concentration of AA. PPE-OB-PEG has a good linear range towards AA from 1.00 × 10-7 to 8.00 × 10-5 mol L-1 and the limit of detection (LOD) is 3.00 × 10-8 mol L-1 (S/N = 3). PPE-OB-PEG have been applied to detect AA in traditional Chinese medicine samples and the results are satisfactory. The experimental results show that PPE-OB-PEG can be used as a fluorescence probe for rapid and sensitive detection of AA.

Life-stage specific transcriptomes of a migratory endoparasitic plant nematode, Radopholus similis elucidate a different parasitic and life strategy of plant parasitic nematodes.

Radopholus similis is an important migratory endoparasitic nematode, severely harms banana, citrus and many other commercial crops. Little is known about the molecular mechanism of infection and pathogenesis of R. similis. In this study, 64761 unigenes were generated from eggs, juveniles, females and males of R. similis. 11443 unigenes showed significant expression difference among these four life stages. Genes involved in host parasitism, anti-host defense and other biological processes were predicted. There were 86 and 102 putative genes coding for cell wall degrading enzymes and antioxidase respectively. The amount and type of putative parasitic-related genes reported in sedentary endoparasitic plant nematodes are variable from those of migratory parasitic nematodes on plant aerial portion. There were no sequences annotated to effectors in R. similis, involved in feeding site formation of sedentary endoparasites nematodes. This transcriptome data provides a new insight into the parasitic and pathogenic molecular mechanisms of the migratory endoparasitic nematodes. It also provides a broad idea for further research on R. similis.

Predictive risk factors for exudative retinal detachment after vitrectomy for proliferative diabetic retinopathy.

This retrospective study investigated the risk factors of exudative retinal detachment (ERD) occurring after vitrectomy performed to treat proliferative diabetic retinopathy (PDR).All patients were treated with vitrectomy for PDR. Patients with history(s) of the following were excluded: ocular surgery (except phacoemulsification combined with intraocular lens implantation or retinal laser photocoagulation); ocular trauma; systemic diseases; ocular diseases; uveitis; scleritis; tumor; congenital ocular disorders; or others.Included were 205 eyes of 169 patients, of whom 18 (8.78%) developed ERD with varying degrees of exudative choroidal detachment after 1 to 3 days. Binary logistic regression showed the following association with the development of ERD: lower serum albumin concentration (P = .001); without intravitreal anti-vascular endothelial growth factor (anti-VEGF) drug injection before vitrectomy (P = .044); and history of phacoemulsification combined with intraocular lens implantation (P = .046). No association was found with gender; age; systolic pressure; diastolic pressure; panretinal photocoagulation; intraocular pressure on the 1st postoperative day; intraocular pressure on the 2nd postoperative day; serum albumin concentration; or blood urea nitrogen.Risk factors for ERD after vitrectomy for PDR include low serum albumin concentration, without history of intravitreal anti-VEGF drug injection before surgery, and a history of phacoemulsification combined with intraocular lens implantation.

Screening of reference genes in real-time PCR for Radopholus similis.

Six candidate reference genes were chosen from the transcriptome database of Radopholus similis using the bioinformatics method, including four conventional reference genes (actin, Eukaryotic translation initiation factor 5A (eIF5A), Tubulin alpha (a-tubulin), ubiquitin (UBI)) and two new candidate reference genes (Ribosomal protein S21 (Rps21) and Serine/threonine protein phosphatase PP1-ß catalytic subunit (ß-PP1)). In addition, a traditional reference gene 18S ribosomal RNA (18S rRNA) obtained from NCBI databases was also added to the analysis. Real-time PCR was used to detect the expression of seven candidate reference genes in six populations of R. similis and four developmental stages (female, male, larva and egg) of a population. The stability of the expression of candidate genes was evaluated by three software programs, BestKeeper, geNorm and NormFinder. The results showed that eIF5A is the most suitable reference gene for gene functional research of different populations, while both Rps21 and eIF5A are the most suitable reference genes for different developmental stages of a population. Therefore, eIF5A is the best reference gene for studying R. similis. However, one defect of this study is that only seven candidate reference genes were analyzed; ideally, more genes should be tested.

Description and molecular analysis of Tylencholaimushelanensis sp. n. from China (Dorylaimida, Tylencholaimidea).

A new species, Tylencholaimushelanensis sp. n., extracted from the rhizosphere soil of unidentified grasses from Helan Mountain, Inner Mongolia, China was identified. The new species is characterized by having a body length of 0.93-1.07 mm with the lip region approximately one-quarter of the body diameter at the posterior end of the neck region wide; female didelphic-amphidelphic; pars proximalis vaginae violin-shaped. Males were not found. SEM observations of the new species were made and a phylogenetic analysis of both the 18S rDNA and the D2-D3 region of 28S rDNA is presented.

Molecular identification and functional characterization of the cathepsin B gene (Ab-cb-1) in the plant parasitic nematode Aphelenchoides besseyi.

The rice white tip nematode, Aphelenchoides besseyi, is widely distributed in rice planting areas worldwide and causes serious economic losses. Cathepsin genes have been demonstrated to have importance in studying the reproduction, development, pathogenicity, and control methods of plant nematodes. In this paper, a novel cathepsin B gene, Ab-cb-1, was found and cloned. The Ab-cb-1 gene was 1347 bp in length and encodes 369 amino acids. The Ab-CB-1 protein contains characteristic occluding loops but no signal peptide. A homology analysis showed that Ab-CB-1 had the highest identity value (64%) to the known amino acid sequence of cathepsin B-like cysteine protease 6 from Toxocara canis. When Ab-cb-1 was expressed in a prokaryotic system, the protein massed approximately 45 kDa and could decompose carrot callus. Ab-cb-1 mRNA was localized in the nematode intestine. The relative expression level of Ab-cb-1 in the A. besseyi Ab-S24 population, which had high reproductivity, was approximately 6.9 times that in the Ab-N10 population, which had low reproductivity, and the difference was significant (p<0.05). The Ab-cb-1 expression level was highest in females; the expression levels in males, juveniles and eggs were 30%, 12.2% and 5% of that in females, respectively, and the differences were significant among all four stages (p<0.05). Nematodes of the Ab-S24 population were treated with Ab-cb-1 dsRNA for 12 h, 24 h, 36 h and 48 h, and their reproduction decreased with increasing time. These results demonstrated that Ab-CB-1 was a digestive enzyme with hydrolytic protease properties and that Ab-cb-1 played an important role in the reproduction of A. besseyi.

Identification and function of FAR protein family genes from a transcriptome analysis of Aphelenchoides besseyi.

Motivation: The rice white tip nematode (RWTN) Aphelenchoides besseyi is a migratory plant parasitic nematode that infects the aboveground parts of plants. Fatty acid- and retinoid-binding (FAR) proteins are nematode-specific proteins that are involved in many important biological processes. Genes encoding FAR proteins have been identified in many species of nematodes, which indicated that nematodes may produce more than one type of FAR protein. The main goal of this study is to find new molecular targets including new far genes that will help control RWTN, and reduce the economic damage caused by RWTN. Results: Two RWTN populations with different levels of pathogenicity and reproduction were sequenced and analyzed with next-generation sequencing. 17 087 transcripts were annotated using six databases and 1696 differentially expressed genes (DEGs) were identified between the two RWTN populations. Seven new Ab-far genes were identified from the transcriptome data of the two RWTN populations which is the first to identify multiple far genes in plant parasitic nematodes. This study is the first to identify far genes in the nervous system of nematodes and the first to report a transcriptome sequencing analysis of different RWTN populations. The results help elucidate the genes related to parasitism and pathogenicity and also contribute to the identification of new target genes and development of new methods to control RWTN. Availability and implementation: Our data are publicly available at Sequence Read Archive (SRA) database and GenBank database. Supplementary information: Supplementary data are available at Bioinformatics online.

A new species of the genus Eudorylaimus Andrássy, 1959 (Nematoda: (Dorylaimida: Qudsianematidae) associated with Picea crassifolia in China.

A new species, Eudorylaimus piceae n. sp., extracted from rhizosphere soil of Picea crassifolia from Inner Mongolia, China was identified. The new species is characterized by these combined characters: body length of 1.03-1.27 mm; lip region distinctly offset; odontostyle 20-22 µm and 1.1-1.4 times the lip region diameter in length; odontophore 1.1-1.2 times the odontostyle length; basal expansion of pharynx occupying 42%-50.5% of the total neck length; genital system didelphic-amphidelphic; vulva transverse; vagina extending inwards 32%-43%; V value averaging more than 60; pars refringens vaginae with two trapezoidal sclerotizations and pars distalis vaginae weakly sclerotized; prerectum 72-107 µm long, 2.3-3.3 times the anal body diameter, and rectum 1-1.6 times the anal body diameter in length; tail conoid and bent ventrally, c' value 1.5-1.8 and males unknown. Phylogenetic analyses based on sequences of 18S small subunit rDNA and the D2-D3 expansion region of 28S rDNA are presented.