[Analysis of HIV-1 genetic subtype and pretreatment drug resistance among men who have sex with men infected with HIV-1 from 19 cities of 6 provinces in China].

Objective: To investigate the distribution of HIV-1 genetic subtypes and pretreatment drug resistance (PDR) among men who have sex with men (MSM) from 19 cities of 6 provinces in China. Methods: From April to November 2019, 574 plasma samples of ART-naive HIV-1 infected MSM were collected from 19 cities in Hebei, Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong provinces, total ribonucleic acid (RNA) was extracted and amplified the HIV-1 pol gene region by nested polymerase chain reaction (PCR) after reverse transcription. Then sequences were used to construct a phylogenetic tree to determine genetic subtypes and submitted to the Stanford drug resistance database for drug resistance analysis. Results: A total of 479 samples were successfully amplified by PCR. The HIV-1 genetic subtypes included CRF01_AE, CRF07_BC, B, CRF55_01B, CRF59_01B, CRF65_cpx, CRF103_01B, CRF67_01B, CRF68_01B and unrecognized subtype, which accounted for 43.4%, 36.3%, 6.3%, 5.9%, 0.8%, 0.8%, 0.4%, 0.4%, 0.2% and 5.5%, respectively. The distribution of genetic subtypes among provinces is statistically different (χ2=44.141, P<0.001). The overall PDR rate was 4.6% (22/479), the drug resistance rate of non-nucleoside reverse transcriptase inhibitors, nucleoside reverse transcriptase inhibitors, and protease inhibitors were 3.5% (17/479), 0.8% (4/479) and 0.2% (1/479), respectively. The PDR rate of recent infections was significantly higher than that of long-term infections (χ2=4.634, P=0.031). Conclusions: The HIV-1 genetic subtypes among MSM infected with HIV-1 from 19 cities of 6 provinces in China are diverse, and the distribution of subtypes is different among provinces. The overall PDR rate is low, while the PDR rate of recent infections was significantly higher than that of long-term infections, suggesting the surveillance of PDR in recent infections should be strengthened.

Species Identification of Sarcosaprophagous Flies Based on Vein Digital Image Analysis.

ABSTRACT: Objective To identify species of common sarcosaprophagous flies based on digital image analysis of veins, in order to provide new idea for fast and accurate species identification of sarcosaprophagous flies in forensic entomology. Methods Random trapping of 226 male and female sarcosaprophagous flies that comprised of 7 common species, including Sarcophaga peregrina, Parasarcophaga ruficornis, Sarcophaga dux, Seniorwhitea reciproca, Bercaea cruentata, Aldrichina grahami, and Synthesiomysia nudiseta with carrion in the field was conducted. The 17 landmarks on the right wing of each fly were digitally processed and the images were analyzed. The effects of allometry were evaluated using a permutation test. Wing shape variations among 7 sarcosaprophagous fly species and female species was analyzed using canonical variate analysis （CVA）. Additionally, cross-validation test was used to evaluate the reliability of classification. Results Among 7 sarcosaprophagous fly species and female species, the effect of allometry had statistical significance （P<0.05）. The CVA results showed that among 7 sarcosaprophagous fly species and female species, differences in the wing shape were significant, and the first two canonical variates accounted for 82.9% and 84.1% of the total variation of vein shape. Vein digital image analysis can be used to separate the 7 common sarcosaprophagous flies, with an overall species identification accuracy of 81.2%-100.0%, and with a species identification accuracy of 75.0%-100.0% to distinguish the female flies of the 7 sarcosaprophagous flies species. Conclusion Vein digital image analysis is a relatively convenient and reliable method for identification of insect species, which can be used for species identification of common sarcosaprophagous flies.

[The combined application of magnetic resonance DTI and 3D-pcASL in differential diagnosis of hyper-acute and acute ischemic cerebral infarction].

Objective: To explore the clinical value of magnetic resonance diffusion tensor imaging (DTI) combined with 3D pseudo continuous arterial spin labeling (3D-pcASL) perfusion imaging in differential diagnosis of hyper-acute and acute ischemic cerebral infarction. Methods: A total of 42 patients with confirmed unilateral ischemic cerebral infarction from February 2015 to October 2017 of Affiliated Yancheng Hospital of Southeast University Medical College were collected. DTI and 3D-pcASL images of hyper-acute (14 patients, group A) and acute (28 patients, group B) ischemic cerebral infarction were retrospectively analyzed. The slice with the biggest lesion on diffusion weighted imaging (DWI) was selected to measure the infarction area (S(DWI)) and abnormal cerebral blood flow (CBF) perfusion area (S(CBF)). Regions of interest (ROIs) were set on the infarction core (IC) area, mismatch area of S(CBF) and S(DWI) (MACD), and their corresponding contralateral regions. The values of CBF, average diffusion coefficient (DC(avg)), and fractional anisotropy (FA) parameter images in these ROIs in patients with both S(CBF)>S(DWI) and lower perfusion in the IC were recorded. The relative values of each infarction lesion to its corresponding contralateral region (rCBF, rFA, and rDC(avg)) were calculated. Differences of each parameter value between the IC, MACD and their corresponding contralateral regions and of the relative values between group A and group B were investigated. Results: The CBF and DC(avg) values in the IC and the CBF value in the MACD were lower than that of their corresponding contralateral regions in both groups (P<0.05). The DC(avg) in the MACD in group A patients and the FA in the IC in group B patients were lower than that in their corresponding contralateral regions (P<0.05). Compared to group A patients, group B patients showed decreased rFA, rDC(avg) in the IC and rFA in the MACD, and increased rDC(avg) and rCBF in the MACD (P<0.05). Receiver operating characteristics (ROC) curve analysis indicated that the best diagnosis cut off values of the rFA and rDC(avg) values in the IC and the rCBF, rFA, and rDC(avg) values in the MACD were 0.890 and 0.541 and 1.139, 0.902 and 0.455, respectively, for identifying two groups. Conclusion: The changes of the CBF, FA, and DC(avg) values and their relative values can be applied to differentially diagnose patients with hyper-acute and acute cerebral infarction, which could provide the basis for selecting more reasonable treatment protocols.

Identification and functional analysis of a novel chorion protein essential for egg maturation in the brown planthopper.

In insect eggs, the chorion has the essential function of protecting the embryo from external agents during development while allowing gas exchange for respiration. In this study, we found a novel gene, Nilaparvata lugens chorion protein (NlChP), that is involved in chorion formation in the brown planthopper, Nilaparvata lugens. NlChP was highly expressed in the follicular cells of female adult brown planthoppers. Knockdown of NlChP resulted in oocyte malformation and the inability to perform oviposition, and electron microscopy showed that the malformed oocytes had thin and rough endochorion layers compared to the control group. Liquid chromatography with tandem mass spectrometry analysis of the eggshell components revealed four unique peptides that were matched to NlChP. Our results demonstrate that NlChP is a novel chorion protein essential for egg maturation in N. lugens, a hemipteran insect with telotrophic meroistic ovaries. NlChP may be a potential target in RNA interference-based insect pest management.

[Comparison of commercial HIV-1 viral load tests by using proficiency test results in China, 2013- 2015].

Objective: To compare the bio-equivalence among commercial HIV-1 viral load tests, including EasyQ HIV-1 v2.0 (EasyQ) from bioMerieux NucliSens of France; VERSANT HIV-1 RNA 3.0 assay (bDNA) from Siemens Healthcare Diagnostics of USA; COBAS AmpliPrep/COBAS TaqMan HIV-1 test (Taqman) from Roche Molecular Diagnosis of USA; Abbott Real Time HIV-1 Kit (M2000) from Abbott Molecular of USA and two domestic HIV-1 viral load test kits (domestic kit) from DaAn Gene Company of Sun Yat-Sen University and Liaoning Bio-Pharmaceutical company of Northeast pharmaceutical group, by using proficiency test results in China from 2013 to 2015. Methods: A total of 2 954 proficiency test results, obtained from 22 positive samples of 6 proficiency tests in 155 laboratories conducted by China CDC were analyzed during 2013-2015. The results from each sample were first logarithmic transformed and then grouped according to the method used, the mean value of logarithmic results was calculated. Subsequently, 22 clusters of mean values were analyzed by Bland-Altman analysis for the consistency, and linear regression analysis for the interdependency. Results: The results indicated that, by taking Taqman as the reference, EasyQ, M2000, bDNA and domestic kit had good consistency (90%-100%) and interdependency. Conclusion: All the viral load tests were bio-equivalent. Moreover, according to the conversion formula derived from domestic proficiency test results, all the viral load results could be converted, which is critical for epidemiological analysis.

[Effect of laboratory referencing on data analysis of community viral load in HIV positive MSM from 15 cities, China].

Objective: To compare the community viral load (CVL) among MSM in 15 cities in China using standardized national reference sources. Methods: The study analyzed the existing database of National Major Science and Technology Project of China. The database was established with serial random survey of MSM HIV CVL among MSM in 15 cities from 2013 to 2015. VL tests were conducted in 15 laboratories with different equipment and methods, including RT-PCR, nucleic acid sequence based amplification (NASBA), branched DNA testing (bDNA) and Abbott M2000 RealTime system (M2000). Based on proficiency test for 15 laboratories conducted by National HIV Reference Laboratory, VL test values detected with EasyQ, bDNA and M2000 were converted and standardized into resultant values of TaqMan 2.0. Software SPSS 17.0 was used to produce descriptive statistics for the dataset. Results: From 2014 to 2015, the 15 testing sites were found to use a number of different viral load detection techniques. In 2014, the community viral load values were (2.38±1.47) and (2.99±1.31) in 15 testing sites, while in 2015 these values were found to be (2.07±1.34) and (2.72±1.19). The measurement of community VL was done using standard benchmarks of ≤200 copies/ml, ≤400 copies/ml and ≤1 000 copies/ml, that were used for reference for now. Conclusion: It is necessary to use standard detection method to improve the comparability of annual results. Using a standardized rate of ≤400 copies/ml or ≤1 000 copies/ml for successful control of VL was found with high stability for the result comparison among different areas.

The ß-N-acetylhexosaminidase gene family in the brown planthopper, Nilaparvata lugens.

ß-N-Acetylhexosaminidases (HEXs) are enzymes that can degrade the chitin oligosaccharides that are produced by the activity of chitinases on chitin in insects. Using bioinformatic methods based on genome and transcriptome databases, 11 ß-N-acetylhexosaminidase genes (NlHexs) in Nilaparvata lugens were identified and characterized. Phylogenetic analysis revealed a six-grouped tree topology. The O-Linked N-acetylglucosaminidase (O-GlcNAcase) group includes NlHex11, which harbours a catalytic domain that differs from that of the other 10 NlHexs. Observations of the expression of NlHexs during different developmental stages revealed that NlHex4 is expressed with periodicity during moulting. Although the tissue-specific expression patterns of most NlHexs were nonspecific, NlHex4 was found to be expressed mainly in the female reproductive system as well as in the integument. RNA interference (RNAi) demonstrated failure to shed the old cuticle only in the nymphs treated with double-stranded RNA (dsRNA) targeting NlHex4, and these nymphs eventually died; no observable morphological abnormalities were found in insects treated with dsRNAs targeting the other 10 NlHexs. Based on this study and our previous analyses, a '5 + 1 + 3' pattern of chitinolytic enzymes is proposed, in which five chitinases, one NlHEX and three chitin deacetylases are required for moulting in N. lugens. A better understanding of chitin metabolism in the hemimetabolous insect, N. lugens, would be achieved by considering three chitinolytic enzyme families: chitinase, chitin deacetylase and ß-N-acetylhexosaminidase.

Quantitative meta-analysis of grey matter anomalies in neuropathic pain.

BACKGROUND: Increasing neuroimaging studies have revealed grey matter (GM) anomalies of several brain regions by voxel-based morphometry (VBM) studies in patients with neuropathic pain. The changes have been suggested to be related to central sensitization. Our aim was to investigate concurrence across VBM studies to identify whether different subtypes of neuropathic pain share a common pathophysiological basis revealed by structural abnormalities. METHODS: A systematic search of VBM studies of patients with neuropathic pain and healthy controls published in PubMed and Embase databases from January 2000 to March 2014 was conducted. A quantitative meta-analysis of whole-brain VBM studies in patients with neuropathic pain compared with healthy controls was performed by means of effect-size signed differential mapping. RESULTS: Ten studies comprising 240 patients with neuropathic pain and 263 healthy subjects were systematically included in the present study. Compared to healthy controls, the patients showed consistent decreased GM in bilateral anterior insula and thalamus, right superior frontal gyrus and left postcentral gyrus, and increased GM in right medial frontal gyrus and right posterior insula. The results remained largely unchanged in the following jackknife sensitivity analysis. CONCLUSIONS: This meta-analysis shows strong evidence of brain GM anomalies within the pain matrix in patients with neuropathic pain compared with healthy subjects. Further studies are needed to determine whether the reported changes are specific to neuropathic pain or whether they may be common to other chronic pain.

Chitinase-like gene family in the brown planthopper, Nilaparvata lugens.

Chitinases are important enzymes required for chitin degradation and reconstruction in insects. Based on a bioinformatics investigation, we identified 12 genes encoding putative chitinase-like proteins, including 10 chitinases (Cht), one imaginal disc growth factor (IDGF) and one endo-ß-N-acetylglucosaminidase (ENGase) in the genome of the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae). These 12 genes were clustered into nine different groups, with 11 in glycoside hydrolase family 18 groups (groups I-VIII) and one in the ENGase group. Developmental and tissue-specific expression pattern analysis revealed that the transcript levels of eight genes peaked periodically during moulting and were mainly expressed in the integument, except NlCht2, NlCht4, NlIDGF and NlENGase. NlCht2, NlIDGF and NlENGase were expressed at all stages with slight periodical changes and mainly expressed in the female reproductive organs in adults, whereas NlCht4 was highly expressed only at the adult stage in the male reproductive organs. Lethal phenotypes were observed in insects challenged by double-stranded RNAs for NlCht1, NlCht5, NlCht7, NlCht9 and NlCht10 during moulting, suggesting their significant roles in old cuticle degradation. NlCht1 was the most sensitive gene, inducing 50% mortality even at 0.01 ng per insect. Our results illustrate the structural and functional differences of chitinase-like family genes and provide potential targets for RNA interference-based rice planthopper management.

Gray matter anomalies in anterior cingulate cortex as a correlate of depressive symptoms in drug-naïve idiopathic restless legs syndrome.

BACKGROUND: Depressive symptoms are frequent in idiopathic restless legs syndrome (RLS). However, little is known, so far, about the neurological basis. The present study aimed to explore the neuroanatomical anomalies in depressed drug-naïve RLS patients using voxel-based morphometry (VBM) analysis. METHODS: We recruited 16 drug-naïve idiopathic RLS patients with depressive symptoms (RLS-D), 18 drug-naïve idiopathic RLS patients without depressive symptoms (RLS-ND), and 18 normal controls. All participants underwent structural MRI scans on a 3-T MR system. The differences in regional gray matter (GM) density were determined across groups by VBM8. Additional regression analysis was used to identify any associations between regional GM density and clinical symptoms. RESULTS: GM density of the bilateral anterior cingulate cortex (ACC) was significantly reduced in RLS-D patients when compared to RLS-ND patients or to the healthy controls. However, there were no significant differences of GM density either when the whole RLS group or the RLS-ND group was compared to healthy controls, respectively. Particularly, we found GM density of right ACC was negatively correlated with the severity and duration of depressive symptoms in RLS-D patients. CONCLUSIONS: Depressive symptoms are associated with GM anomalies in ACC in patients with RLS. We propose that ACC is perhaps an important neuroimaging marker for facilitating treatment strategies in patients with RLS when assessing depressive symptoms.

Chitin deacetylase family genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae).

Chitin deacetylases (CDAs) are enzymes required for one of the pathways of chitin degradation, in which chitosan is produced by the deacetylation of chitin. Bioinformatic investigations with genomic and transcriptomic databases identified four genes encoding CDAs in Nilaparvata lugens (NlCDAs). Phylogenetic analysis showed that insect CDAs were clustered into five major groups. Group I, III and IV CDAs are found in all insect species, whereas the pupa-specific group II and gut-specific group V CDAs are not found in the plant-sap/blood-sucking hemimetabolous species from Hemiptera and Anoplura. The developmental and tissue-specific expression patterns of four NlCDAs revealed that NlCDA3 was a gut-specific CDA, with high expression at all developmental stages; NlCDA1, NlCDA2 and NlCDA4 were highly expressed in the integument and peaked periodically during every moulting, which suggests their roles in chitin turnover of the insect old cuticle. Lethal phenotypes of cuticle shedding failure and high mortality after the injection of double-stranded RNAs (dsRNAs) for NlCDA1, NlCDA2 and NlCDA4 provide further evidence for their functions associated with moulting. No observable morphological and internal structural abnormality was obtained in insects treated with dsRNA for gut-specific NlCDA3.

Genome-wide screening for components of small interfering RNA (siRNA) and micro-RNA (miRNA) pathways in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae).

The brown planthopper (BPH), Nilaparvata lugens, is a major rice pest in Asia, and accumulated evidence indicates that this species is susceptible to RNA interference (RNAi); however, the mechanism underlying RNAi and parental RNAi has not yet been determined. We comprehensively investigated the repertoire of core genes involved in small interfering RNA (siRNA) and micro-RNA (miRNA) pathways in the BPH by comparing its newly assembled transcriptome and genome with those of Drosophila melanogaster, Tribolium castaneum and Caenorhabditis elegans. Our analysis showed that the BPH possesses one drosha and two Dicer (dcr) genes, three dsRNA-binding motif protein genes, two Argonaute (ago) genes, two Eri-1-like genes (eri-1), and a Sid-1-like gene (sid-1). Additionally, we report for first time that parental RNAi might occur in this species, and siRNA pathway and Sid-1 were required for high efficiency of systemic RNAi triggered by exogenous dsRNA. Furthermore, our results also demonstrated that the miRNA pathway was involved in BPH metamorphosis as depletion of the ago1 or dcr1 gene severely impaired ecdysis. The BPH might be a good model system to study the molecular mechanism of systemic RNAi in hemimetabolous insects, and RNAi has potential to be developed to control this pest in agricultural settings.

Voxel-wise meta-analysis of gray matter abnormalities in idiopathic Parkinson's disease.

Structural neuroimaging studies on idiopathic Parkinson's disease (IPD) with voxel-based morphometry (VBM) yielded variable and conflicting findings. A systematic review of VBM studies of patients with IPD and healthy control (HC) subjects published in PubMed, ISI Web of Science, Embase, and Medline databases from 1995 to 25 October 2010 was conducted. Coordinates were extracted from clusters of significant gray matter (GM) difference between patients with IPD and HC subjects. Meta-analysis was performed using signed differential mapping. A total of 17 VBM studies involving 498 patients with IPD and 375 HC subjects met the inclusion criteria. A significant regional GM volume decrease was detected in the left inferior frontal gyrus (BA47) extending to the left superior temporal gyrus (BA38) and the left insula (BA13) of patients with IPD compared with HC subjects. The findings of this study remain largely unchanged in quartile and jackknife sensitivity analyses and in subgroup analyses. Robust GM reductions in the inferior frontal/orbitofrontal gyrus (BA47) are implicated in IPD, and the reductions may be related to the mediation of the non-motor IPD symptoms, such as cognitive, emotional, and autonomic functions. Further studies must be conducted to determine whether the findings are specific to all IPD subtypes or different from the atypical Parkinsonism.