[Expression level of Wilms' tumor 1 gene and its correlation with clinical features in patients with myeloproliferative neoplasms].

Objective: To investigate the expression level of WT1 gene in patients with classical Philadelphia chromosome (Ph)-negative myeloproliferative neoplasms (MPN) and its correlation with clinical features. Methods: A retrospective study included 252 patients with newly diagnosed MPN in Zhongnan Hospital of Wuhan University from January 2015 to March 2023, including 128 males and 124 females, aged［M（Q1，Q3）］62 (53, 69) years. The WT1-positive group (n=93) and the WT1-negative group (n=159) were split based on the level of WT1 gene expression, and the variations in clinical indicators between the two groups were compared. Its levels of expression in each subtype and its relationships to thrombotic events and clinically significant variables were analyzed. As of March 31, 2023, the follow-up period [M (Q1, Q3)] was 12.0(6.5,21.0)months. The risk factors of thrombosis in MPN patients were analyzed by using the logistic regression analysis. Results: The WT1 gene expression level in the overall bone marrow samples of 252 patients with newly diagnosed MPN was 0.30% (0.10%, 1.10%). The expression level in primary myelofibrosis (PMF) patients was 1.45% (0.41%, 3.24%), which was higher than 0.15% (0.02%, 0.32%), 0.37% (0.16%, 1.09%) in essential thrombocythemia (ET) and polycythemia vera (PV) patients (both P<0.05). Positive correlations were found between WT1 gene expression levels and JAK2V617F gene mutation load, RDW, MPV (r=0.478, 0.346, 0.236, all P<0.01). While negative correlations between WT1 gene expression levels and PLT, LYM, PTTA, LDH were found (r=-0.339, -0.170, -0.206, -0.388, all P<0.01). Patients in the WT1-positive group exhibited a higher percentage of somatic symptoms, splenomegaly, positive JAK2V617F gene mutation, and higher levels of RDW, LDH, NEUT, and MPV compared to the WT1-negative group. In contrast, the proportion of triple-negative (negative for all three hot mutations of JAK2V617F, CALR and MPL) was lower, and the levels of PLT, LYM and PTTA were lower (all P<0.05). The thrombotic event rates of WT1-positive group and WT1-negative group were 32.3% (30/93) and 32.1% (51/159), respectively, and the difference was not statistically significant (P=0.883). Logistic regression analysis showed that male (OR=2.41,95%CI:1.02-5.71,P=0.046) and positive JAK2V617F gene mutation (OR=3.96,95%CI:1.50-10.42,P=0.005) were risk factors for thrombotic events in ET patients. Conclusions: WT1 gene expression is elevated in PMF patients and correlated with indicators of disease progression and transformation in MPN patients. It can be utilized as an auxiliary diagnostic indicator for classical MPN staging but is not correlated with the incidence of thrombotic events. Male and positive JAK2V617F gene mutation are risk factors for thrombotic events in ET patients.

The bipolar charge plasma spectrometer (BCPS) based on the 2π-field-of-view double-channel electrostatic analyzer.

In this paper, we propose a bipolar charge plasma spectrometer based on the double-channel electrostatic analyzer for simultaneously measuring thermal ions and electrons with a 2π hemispherical field-of-view. Both ions and electrons within the wide field-of-view enter the spectrometer, pass through the variable geometric factor channel, and are then separated by the double-channel electric fields. Two microchannel plates are accommodated at the exit of the analyzer for ion and electron detection. The main performance of the spectrometer has been obtained from on-ground calibration. With the electrostatic deflectors and the cylindrically symmetric structure, the spectrometer provides simultaneous measurements of thermal ion and electron velocity distributions with a shared field-of-view of 360° (azimuth angle) by 90° (elevation angle) and a broad energy range for both ions and electrons. The ion analyzer constant and the electron analyzer constant are 11.1 and 9.7, respectively. The detecting energy range of 33.3-44.4 keV for ions and 29.1-38.8 keV for electrons can be obtained by using the sweeping electrostatic analyzer voltage range of 3-4000 V. The ion and electron energy resolutions are 9.6% and 6.1%, respectively. The variable geometric factor function provides a large geometric factor adjusting range for both ion and electron measurements by two orders of magnitude, which fulfills the requirements of a large dynamic flux range for simultaneous measurements of space thermal plasma in the solar wind and magnetosphere.

Performance and simulated moment uncertainties of an ion spectrometer with asymmetric 2π field of view for ion measurements in space.

Space plasma instruments provide 3D particle velocity distribution functions. Because of telemetry limitations, these cannot be transmitted in high time resolution and the plasma needs to be characterized by moments of the velocity distribution function. These moment uncertainties have vital effects on the reliability and accuracy of onboard plasma moments. We assess the measurement accuracy for magnetosheath and solar wind ions using an ion spectrometer with an asymmetric field of view designed for the all-sky measurement of low-energy ions in the magnetosheath and solar wind. We focus on moment uncertainties for the ideal spectrometer, not considering the background counts, which may have considerable effects on the uncertainties in real life. To obtain number density, bulk velocity, and temperature, different orders of moments are integrated assuming a Maxwellian velocity distribution. Based on the design specifications, we use simulations to estimate systematic and random errors for typical plasma conditions. We find that the spectrometer resolution is adequate for determining the density of solar wind (∼7% error) and magnetosheath ions (∼4% error). The resolution is also adequate for determining the temperature of solar wind (∼10% error) and magnetosheath ions (∼2% error). For high speed flows with a bulk velocity of 750 km/s and a temperature of 20 eV, the maximum density and temperature errors become 9% and 7%, respectively. The bulk velocity errors are less than 2% for all cases. The contributions of heavy ions to the systematic errors are less than 5% for magnetosheath ions and less than 8% for solar wind ions.

[Risk factors analyses for lateral neck lymph node metastasis in papillary thyroid carcinoma].

Objective:To study the risk factors for the lateral neck lymph node metastasis(LNM) in patients with papillary thyroid carcinoma(PTC).Method:Total 145 cases of papillary thyroid carcinoma patients were reviewed. These patients were divided into the following 2 groups: a lateral LNM-positive group and a lateral LNM-negative group.Clinical factors and tumor ultrasound characteristics were recorded, and the final pathology results were recorded as well. Result: Univariate and multivariate analyses indicated that primary tumor size(pathological diameter) >1.0 cm, upper portion location and lymph node metastasis in central neck were the three risk factors for lateral LNM of PTC(OR=6.250,4.881,5.222,P<0.05). The lymph node metastasis in the lateral neck was not correlated obviously with gender, age, capsuleinvasion, multifocality and tumor ultrasonic characteristics(including illdefined and irregular in shape, calcification, hypervascularized). The probability of lateral LNM was then calculated according to the number of risk factors present in each of the 145 patients, using the three factors. When two or more factors were present, the probability increased up to 38.1 %(95%CI23-53).Conclusion: The present study provides that lateral LNM occurs mainly in patients, with a tumor size(pathological diameter) diameter>1.0 cm, with the primary tumor located in the upper part of the lobe and positive central compartment lymph node metastasis. The risk increased up when at least two of the three factors were present. So it needs to pay attention to the high risk factors seriously which effect lateral neck lymph node metastasis in patients with PTC, combined with ultrasonic characteristics of lateral cervical lymph node or more meticulous evaluations, to establish the lymph node dissection individually.

[Clinical analysis of 234 esophageal foreign bodies].

Objective: To explore the clinical characteristics and treatment methods of esophageal foreign body. Method: The clinical data of 234 patients with esophageal foreign bodies admitted to our department from January 2015 to August 2018 were retrospectively analyzed, including course time, foreign body types, surgical methods, imaging manifestations and treatment related complications. Result: The diagnosis of esophageal foreign bodies was confirmed by esophageal CT or esophageal barium meal X-ray examination before operation in 234 patients. Course time varied from 3 hours to 7 days, and the jujube nucleus was the most common food-borne foreign body.223 patients underwent esophagoscopic exploration and foreign body removal under general intravenous anesthesia, 11 of them had no definite esophageal foreign body, 22 had esophageal perforation and periesophagitis. After removal of foreign body, the nasogastric feeding tube was inserted. About 10 days later, the nasogastric feeding tube was removed when they got healthy. Nine cases underwent cervical abscess incision and drainage under general anesthesia. The average postoperative hospital day was 11 days. Conclusion: The rigid esophagoscopy is a safe and effective method for the esophageal foreign bodies. And neck abscess incision must be necessary,when they suffered from esophageal perforation with neck abscess and other serious complications.

[The diagnosis and treatment of neck abscess and mediastinal abscess following esophageal perforation induced by esophageal foreign body].

Objective:The aim of this study is to discuss the clinical presentation, imaging examination and treatment of neck abscess and mediastinal abscess following esophageal perforation induced by esophageal foreign body. Method:Six patients all underwent lateral neck incision and drainage of neck abscess. Simultaneously, mediastinal abscess drainage was performed in 3 cases with mediastinal abscess. Result:All 6 patients were cured. The median time of extraction of gastric tube was 11 days (7-30 days). All patients were not treated with tracheotomy. Conclusion:Neck and chest CT should be done as soon as possible for suspected patients to definite the location of foreign body and the relationship between foreign body and surrounding structures. Patient who were suspected with neck abscess and mediastinal abscess should undergo surgery to remove foreign body and drain the infectious deep neck and mediastinal spaces as early as possible.

[Risk factors of central neck lymph node metastasis in cN0 papillary thyroid carcinoma].

Objective:To investigate the risk factors of central neck lymph node metastasis (CNLNM) in cN0 papillary thyroid carcinoma. Method:Retrospective analysis of 114 patients with papillary thyroid carcinoma in stage of cN0 who underwent primary treatment. Collected the clinical and pathological data, used the univariate and multivariate analysis to investigate the risk factors of central neck lymph node metastasis and high volume central neck lymph node metastasis.Result:In the univariate analysis, age (48.2% in<45 years vs 27.6% in≥45 years), multifocal (51% with vs 27.7% without), nodular goiter (58.8% with vs 28.7% without), showed significant difference in prevalence of CNLNM. Age (14.3% in<45 years vs 1.7% in≥45 years), tumor size (19.2% >1 cm vs 4.5%≤1 cm) showed significant difference in prevalence of high volume CNLNM. Inmultivariate analysis, age (RR= 0.304), multifocal (RR= 3.637) and nodular goiter (RR= 4.132) showed the independent risk factor of CNLNM.Conclusion:For cN0 patients with thyroid papillary carcinoma, if the age is younger than 45 years old, the tumor is multifocal, or associatedwith nodular goiter, the surgery should be more aggressive in the prophylactic central neck dissection.

[Primary neuroendocrine carcinoma of the tonsil: two case reports and review of the literature].

Neuroendocrine carcinoma(NEC) of the tonsil is extremely rare. Only a few cases of this tumor have been reported so far. In this article, we described two patients presented with abnormal sensation of throat or throat pain. The pathological examination were NEC. With review of the literatures, the clinical characteristics were summarized and therapeutic strategies were discussed.

Existence of species complex largely reduced barcoding success for invasive species of Tephritidae: a case study in Bactrocera spp.

Fruit flies in the family Tephritidae are the economically important pests that have many species complexes. DNA barcoding has gradually been verified as an effective tool for identifying species in a wide range of taxonomic groups, and there are several publications on rapid and accurate identification of fruit flies based on this technique; however, comprehensive analyses of large and new taxa for the effectiveness of DNA barcoding for fruit flies identification have been rare. In this study, we evaluated the COI barcode sequences for the diagnosis of fruit flies using 1426 sequences for 73 species of Bactrocera distributed worldwide. Tree-based [neighbour-joining (NJ)]; distance-based, such as Best Match (BM), Best Close Match (BCM) and Minimum Distance (MD); and character-based methods were used to evaluate the barcoding success rates obtained with maintaining the species complex in the data set, treating a species complex as a single taxon unit, and removing the species complex. Our results indicate that the average divergence between species was 14.04% (0.00-25.16%), whereas within a species this was 0.81% (0.00-9.71%); the existence of species complexes largely reduced the barcoding success for Tephritidae, for example relatively low success rates (74.4% based on BM and BCM and 84.8% based on MD) were obtained when the sequences from species complexes were included in the analysis, whereas significantly higher success rates were achieved if the species complexes were treated as a single taxon or removed from the data set - BM (98.9%), BCM (98.5%) and MD (97.5%), or BM (98.1%), BCM (97.4%) and MD (98.2%).

Comparative studies on species identification of Noctuoidea moths in two nature reserve conservation zones (Beijing, China) using DNA barcodes and thin-film biosensor chips.

Rapid and accurate identification of species is required for the biological control of pest Noctuoidea moths. DNA barcodes and thin-film biosensor chips are two molecular approaches that have gained wide attention. Here, we compare these two methods for the identification of a limited number of Noctuoidea moth species. Based on the commonly used mitochondrial gene cytochrome c oxidase I (the standard DNA barcode for animal species), 14 probes were designed and synthesized for 14 species shared by two national nature reserves in Beijing and Hebei, China. Probes ranged in length from 18 to 27 bp and were designed as mismatch probes to guarantee that there were at least three base differences between the probe and nontarget sequences. The results on the chip could be detected by the naked eye without needing special equipment. No cross-hybridizations were detected although we tested all probes on the 14 target and 24 nontarget Noctuoidea species. The neighbour-joining tree of the 38 species based on COI sequences gave 38 highly supported independent groups. Both DNA barcoding and thin-film biosensor chips, based on the COI gene, are able to accurately identify and discriminate the 14 targeted moth species in this study. Because of its speed, high accuracy and low cost, the thin-film biosensor chip is a very practical means of species identification. Now, a more comprehensive chip will be developed for the identification of additional Noctuoidea moths for pest control and ecological protection.

A fuzzy-set-theory-based approach to analyse species membership in DNA barcoding.

Reliable assignment of an unknown query sequence to its correct species remains a methodological problem for the growing field of DNA barcoding. While great advances have been achieved recently, species identification from barcodes can still be unreliable if the relevant biodiversity has been insufficiently sampled. We here propose a new notion of species membership for DNA barcoding-fuzzy membership, based on fuzzy set theory-and illustrate its successful application to four real data sets (bats, fishes, butterflies and flies) with more than 5000 random simulations. Two of the data sets comprise especially dense species/population-level samples. In comparison with current DNA barcoding methods, the newly proposed minimum distance (MD) plus fuzzy set approach, and another computationally simple method, 'best close match', outperform two computationally sophisticated Bayesian and BootstrapNJ methods. The new method proposed here has great power in reducing false-positive species identification compared with other methods when conspecifics of the query are absent from the reference database.

Estimating sample sizes for DNA barcoding.

Sample size has long been one of the basic issues since the start of the DNA barcoding initiative and the global biodiversity investigation. As a contribution to resolving this problem, we propose a simple resampling approach to estimate several key sampling sizes for a DNA barcoding project. We illustrate our approach using both structured populations simulated under coalescent and real species of skipper butterflies. We found that sample sizes widely used in DNA barcoding are insufficient to assess the genetic diversity of a species, population structure impacts the estimation of the sample sizes, and hence will bias the species identification potentially.

BPSI2.0: a C/C++ interface program for species identification via DNA barcoding with a BP-neural network by calling the Matlab engine.

BP-Species Identification (BPSI2.0) is a computer program that performs species identification by training a Back-Propagation Neural Network. A short DNA barcoding segment is used as input for training a three-layer BP network. The trained network can assign an unknown query sequence to a known species in the user's database, and provide the corresponding subvector value of the output vector as a relative probability value.

Inferring species membership using DNA sequences with back-propagation neural networks.

DNA barcoding as a method for species identification is rapidly increasing in popularity. However, there are still relatively few rigorous methodological tests of DNA barcoding. Current distance-based methods are frequently criticized for treating the nearest neighbor as the closest relative via a raw similarity score, lacking an objective set of criteria to delineate taxa, or for being incongruent with classical character-based taxonomy. Here, we propose an artificial intelligence-based approach - inferring species membership via DNA barcoding with back-propagation neural networks (named BP-based species identification) - as a new advance to the spectrum of available methods. We demonstrate the value of this approach with simulated data sets representing different levels of sequence variation under coalescent simulations with various evolutionary models, as well as with two empirical data sets of COI sequences from East Asian ground beetles (Carabidae) and Costa Rican skipper butterflies. With a 630-to 690-bp fragment of the COI gene, we identified 97.50% of 80 unknown sequences of ground beetles, 95.63%, 96.10%, and 100% of 275, 205, and 9 unknown sequences of the neotropical skipper butterfly to their correct species, respectively. Our simulation studies indicate that the success rates of species identification depend on the divergence of sequences, the length of sequences, and the number of reference sequences. Particularly in cases involving incomplete lineage sorting, this new BP-based method appears to be superior to commonly used methods for DNA-based species identification.

Species status and phylogeography of two closely related Coptolabrus species (Coleoptera: Carabidae) in South Korea inferred from mitochondrial and nuclear gene sequences.

We investigated the species status and intraspecific phylogeography in South Korea of two ground beetle species, Coptolabrus jankowskii and Coptolabrus smaragdinus (Coleoptera: Carabidae), using statistical parsimony networks and nested clade analyses based on sequences from the mitochondrial cytochrome oxidase subunit I (COI) and nuclear phosphoenolpyruvate carboxykinase (PepCK) and wingless (Wg) genes. Although traditional parsimony tree construction generally failed to resolve interspecific relationships and construct biologically meaningful genealogies, analysis using statistical parsimony networks yielded statistically significant inter- and intraspecific genealogical structures. We found that although these two species represent a notable case of trans-species polymorphisms in both mitochondrial and nuclear gene sequences, their status as separate species was evidenced by the nonrandom association between species and nested clades at various nesting levels. The exceptional occurrence of shared identical or very similar COI sequences was considered to be the result of introgressive hybridization. In addition, range expansion and fragmentation events across the Korean Peninsula and adjacent islands were inferred from nested clade phylogeographical analyses. The COI gene revealed the geographical divergence of major eastern and western clades and historical biogeographical events within each major clade, whereas the nuclear PepCK gene, which did not reveal corresponding east-west clades, indicated past fragmentation and range expansion across wide areas that may have been the result of older biogeographical events. Thus, phylogeographical inferences drawn from analyses of mitochondrial and nuclear genes can reveal different and potentially complementary information about phylogeographical processes.

Cloning and expression of the VP2 gene of an infectious bursal disease virus.

A serotype I infectious bursal disease virus (IBDV) strain HZ96 was isolated in Hangzhou, China, in 1996 and attenuated by adaptation to chicken embryo fibroblast cells. The VP2 gene of strain HZ96 was amplified by reverse transcription-polymerase chain reaction, cloned, and sequenced. Compared with the VP2 sequences of other IBDV strains, HZ96 is most related to two attenuated strains, CU-1 and PBG98, and two attenuated Chinese strains, Harbin and CJ801bkf. HZ96 shares nucleotide sequence homology 98.9% with CU-1 and PBG98, 98.5% with Harbin, and 98.6% with CJ801bkf. Most of the sequence variations observed between HZ96 and other strains are located in the middle variable region from nucleotides 637 to 996. Similar to other attenuated IBDVs, HZ96 has unique substitutions at residues 279 (Asp to Asn) and 284 (Ala to Thr), suggesting that these two substitutions may be directly related to adaptation of the virus to cell culture and attenuation of its virulence. As part of our effort to develop a submit vaccine for IBDV, the VP2 gene of HZ96 was cloned into a heat-inducible expression vector and expressed in Escherichia coli system. A protein band with expected molecular weight of 52 kD was detected by direct protein staining and western blotting.