A member of the HSP90 family from ovine Babesia in China: molecular characterization, phylogenetic analysis and antigenicity.

Heat shock protein 90 (HSP90) is a key component of the molecular chaperone complex essential for activating many signalling proteins involved in the development and progression of pathogenic cellular transformation. A Hsp90 gene (BQHsp90) was cloned and characterized from Babesia sp. BQ1 (Lintan), an ovine Babesia isolate belonging to Babesia motasi-like group, by screening a cDNA expression library and performing rapid amplification of cDNA ends. The full-length cDNA of BQHsp90 is 2399 bp with an open reading frame of 2154 bp encoding a predicted 83 kDa polypeptide with 717 amino acid residues. It shows significant homology and similar structural characteristics to Hsp90 of other apicomplex organisms. Phylogenetic analysis, based on the HSP90 amino acid sequences, showed that the Babesia genus is clearly separated from other apicomplexa genera. Five Chinese ovine Babesia isolates were divided into 2 phylogenetic clusters, namely Babesia sp. Xinjiang (previously designated a new species) cluster and B. motasi-like cluster which could be further divided into 2 subclusters (Babesia sp. BQ1 (Lintan)/Babesia sp. Tianzhu and Babesia sp. BQ1 (Ningxian)/Babesia sp. Hebei). Finally, the antigenicity of rBQHSP90 protein from prokaryotic expression was also evaluated using western blot and enzyme-linked immunosorbent assay (ELISA).

Integrated physical, genetic and genome map of chickpea (Cicer arietinum L.).

Physical map of chickpea was developed for the reference chickpea genotype (ICC 4958) using bacterial artificial chromosome (BAC) libraries targeting 71,094 clones (~12× coverage). High information content fingerprinting (HICF) of these clones gave high-quality fingerprinting data for 67,483 clones, and 1,174 contigs comprising 46,112 clones and 3,256 singletons were defined. In brief, 574 Mb genome size was assembled in 1,174 contigs with an average of 0.49 Mb per contig and 3,256 singletons represent 407 Mb genome. The physical map was linked with two genetic maps with the help of 245 BAC-end sequence (BES)-derived simple sequence repeat (SSR) markers. This allowed locating some of the BACs in the vicinity of some important quantitative trait loci (QTLs) for drought tolerance and reistance to Fusarium wilt and Ascochyta blight. In addition, fingerprinted contig (FPC) assembly was also integrated with the draft genome sequence of chickpea. As a result, ~965 BACs including 163 minimum tilling path (MTP) clones could be mapped on eight pseudo-molecules of chickpea forming 491 hypothetical contigs representing 54,013,992 bp (~54 Mb) of the draft genome. Comprehensive analysis of markers in abiotic and biotic stress tolerance QTL regions led to identification of 654, 306 and 23 genes in drought tolerance "QTL-hotspot" region, Ascochyta blight resistance QTL region and Fusarium wilt resistance QTL region, respectively. Integrated physical, genetic and genome map should provide a foundation for cloning and isolation of QTLs/genes for molecular dissection of traits as well as markers for molecular breeding for chickpea improvement.

[Retracted] A disintegrin and metallproteinase 15 knockout decreases migration of fibroblast­like synoviocytes and inflammation in rheumatoid arthritis.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that certain of the western blotting assay data shown in Figs. 1B and 5A and the histological data shown in Fig. 6C were strikingly similar to data appearing in different form in other articles written by different authors at different research institutes that had either already been published elsewhere prior to the submission of this paper to Molecular Medicine Reports, or were under consideration for publication at around the same time. In view of the fact that certain of these data had already apparently been published previously, the Editor of Molecular Medicine Reports has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 11: 4389­4396, 2015; DOI: 10.3892/mmr.2015.3302].

Salinization of groundwater in shale gas extraction area in the Sichuan Basin, China: Implications for water protection in shale regions with well-developed faults.

The expanding growth of shale gas development has sparked global concern over water-related environmental issues. However, research on groundwater contamination in shale gas areas in China remains limited, impeding environmentally friendly industry practices. To address this gap, we investigated the Wufeng-Longmaxi shale region in the Sichuan Basin, encompassing both operational and prospective shale gas extraction sites, to assess the effects of shale gas operations on shallow groundwater quality. We found there was no significant correlation between groundwater quality and the minimum distance from the shale gas well pads, and some groundwater samples located far from shale gas well pads, rather than those close to pads, were salinized. These findings suggest minimal impacts from shale gas drilling and hydraulic fracturing. The salinized groundwater samples are characterized by high salinity levels and ion concentrations, and are located near fault zones. The primary source of shallow groundwater salinization was derived from the Triassic formation brines confirmed through the assessment of the sensitivity and conservative mixing models. Faults in the study area were identified as pathways for the upward migration of Triassic brines, evidenced by the proximity of salinized samples to fault zones. However, further investigation is required to ascertain whether shale gas extraction activities have induced the migration of formation brines. The occurrence and reactivation of faults, induced by microseismic activities, may pose an increased risk of groundwater contamination in tectonically complex fault zones during shale gas extraction. Therefore, it is imperative to enhance extraction strategies and technologies, particularly in shale regions with well-developed faults, such as optimizing well placement regulation, controlling hydraulic fracturing scale, and strengthening environmental monitoring. By shedding light on potential environmental ramifications of shale gas extraction, especially in fault-rich regions, our study informs water protection strategies and the sustainable advancement of the shale gas industry.

Identification of a novel protein Hq023 of the hard tick Haemaphysalis qinghaiensis and preliminary evaluation of its analgesic effect in mice model.

Tick saliva contains a range of critical biological molecules which could inhibit host defenses and guarantee their food supply. Hq023, a novel cDNA sequence, was cloned from a cDNA library constructed from salivary glands of partially-engorged Haemaphysalis qinghaiensis. Hq023 has an open reading frame (ORF) of 408 bp coding a protein containing 135 amino acid residues with a molecular mass of 15 kDa. Database homology showed that Hq023 protein was structurally similar to a natural toxin U33-theraphotoxin-Cg1c from the Chinese tarantula Chilobrachys guangxiensis. A recombinant protein was expressed with the novel cDNA in a prokaryotic system and its analgesic effect was evaluated in mice model. Both tail immersion and hot-plate tests uncovered an antinociceptive activity, while in the acetic acid-induced writhing test this effect was not observed. These results indicated that the novel recombinant protein Hq023 (rHq023) probably possessed a central antinociceptive activity. Finding of the novel protein might pave a new avenue for the development of tick-derived analgesics.

Promoter methylation status of the tumor suppressor gene SOX11 is associated with cell growth and invasion in nasopharyngeal carcinoma.

BACKGROUND: The transcription factor SOX11 is one of members of the SRY box-containing (SOX) family emerging as important transcriptional regulators. In recent years, up-regulation of SOX11 has been detected in various types of solid tumors. In this study, the effects of promoter methylation of the SOX11 gene on SOX11 expression and cell growth and invasion of nasopharyngeal carcinoma were investigated. METHODS: In this study,methylation-specific PCR and real time quantitative PCR have been applied to investigate the effect of promoter methylation of the SOX11 gene on SOX11 expression in the nasopharyngeal carcinoma and chronic inflammation tissues. The nasopharyngeal carcinoma cell line (CNE2) was treated with 5-aza-2'-deoxycytidine. The effect of promoter methylation of SOX11 on growth and invasion of nasopharyngeal carcinoma cells was detected with MTT test and Boyden chamber Matrigel invasion assay. RESULTS: No or weak expression of SOX11 mRNA was detected in the nasopharyngeal carcinoma tissues of SOX11 gene promoter methylation. Strong expression of SOX11 mRNA was detected in the nasopharyngeal carcinoma tissues of SOX11 gene promoter unmethylation and chronic inflammation tissues of pharynx nasalis. SOX11 mRNA and protein were re-expressed, SOX11 gene was demethylated, and growth and invasion of cells were inhibited in CNE2 cell line after 5-aza-2'-deoxycytidine treatment. CONCLUSIONS: The results of the study indicate that expression of SOX11 mRNA and protein were related to SOX11 gene methylation status. SOX11 gene methylation may be plays a role in growth and invasion of nasopharyngeal carcinoma cells.

Comparison of Outcomes Between Early and Late Tracheostomy.

BACKGROUND: The timing of tracheostomy in ventilated patients remains controversial. This study aimed to compare the effect of early tracheostomy (≤7 d) with late tracheostomy (>7 d) on the prognosis of patients requiring prolonged mechanical ventilation. METHODS: This was a retrospective observational cohort study. The data of 175 patients who received tracheostomy at the ICU between January 1, 2015-July 31, 2022, were collected. Patients were excluded from the study if medical records were incomplete or they underwent tracheostomy as part of a planned operation procedure. One-to-one propensity score matching was used to correct the baseline characteristics between the early and late tracheostomy groups. The treatment process and outcomes were compared between the two groups. The primary outcome was the incidence of ventilator-associated pneumonia (VAP) between groups. RESULTS: After propensity score matching, 88 subjects were included in the analysis. Compared with the late tracheostomy group, the incidence of VAP, hospital length of stay, sedation-free days, ventilator-free days, and ICU-free days were longer in the early tracheostomy group. There were no significant differences in the 90-d mortality between the two groups. CONCLUSIONS;: Early tracheostomy can reduce the occurrence of complications for ICU patients.

Tracing the variation of dissolved organic matter in the two-stage anoxic/aerobic process treating swine wastewater using fluorescence excitation-emission matrix with parallel factor analysis.

Swine wastewater has become one of the main agricultural pollution sources. Quantitative characterization of dissolved organic matter (DOM) is often used in various water bodies, but there are few studies on DOM analysis of swine wastewater. In this study, swine wastewater was treated by a step-feed two-stage anoxic/aerobic (SF-A/O/A/O) process. By using parallel factor (PARAFAC) analysis of fluorescence excitation-emission matrix (EEM), the main components of swine wastewater were aromatic protein-like substances (C1), tryptophan-like substances (C2), fulvic acid-like/humic-like substances (C3) and humic-like substances (C4). Protein-like substances were degraded significantly, while humic-like substances were difficult to be utilized by microorganisms. Fluorescence spectral indexes showed that the characteristics of endogenous input and humus were enhanced. Moreover, several significant correlations between DOM components, fluorescence spectral indexes and water quality indexes were observed. These findings help to understand the biochemical role and the impact of DOM in water quality monitoring and control of swine wastewater.

Long non­coding RNA NEAT1 promotes mouse granulosa cell proliferation and estradiol synthesis by sponging miR­874­3p.

It has been reported that long non-coding RNA nuclear-enriched abundant transcript 1 (NEAT1) is involved in follicular growth and multiple ovarian diseases, but not the physiological function of NEAT1 in mouse granulosa cells (mGCs). Therefore, the aim of the present study was to investigate the biological roles and regulatory mechanisms of NEAT1 in mGCs. The biological effects of NEAT1 on mGCs proliferation, apoptosis, production of 17ß-Estradiol (E2) and progesterone (P4) were investigated using MTS, flow cytometry and enzyme-linked immunosorbent assays, respectively. The association between NEAT1 and microRNA (miR)-874-3p was verified using luciferase reporter assay and RNA immunoprecipitation analysis. The results demonstrated that the knockdown of NEAT1 in mGC cells significantly promoted mGCs cell proliferation, inhibited apoptosis and increased the production of E2 and P4 in mGCs. The interference-mediated effect of NEAT1 on mGCs could be partially reversed by the downregulation of miR-874-3p. Overall, these results indicated that NEAT1 served as a competing endogenous RNA by competitively binding with miR-874-3p, thereby modulating mGCs proliferation and the production of E2 and P4 in mGCs.

Age-related hearing loss accelerates the decline in fast speech comprehension and the decompensation of cortical network connections.

Patients with age-related hearing loss face hearing difficulties in daily life. The causes of age-related hearing loss are complex and include changes in peripheral hearing, central processing, and cognitive-related abilities. Furthermore, the factors by which aging relates to hearing loss via changes in auditory processing ability are still unclear. In this cross-sectional study, we evaluated 27 older adults (over 60 years old) with age-related hearing loss, 21 older adults (over 60 years old) with normal hearing, and 30 younger subjects (18-30 years old) with normal hearing. We used the outcome of the upper-threshold test, including the time-compressed threshold and the speech recognition threshold in noisy conditions, as a behavioral indicator of auditory processing ability. We also used electroencephalography to identify presbycusis-related abnormalities in the brain while the participants were in a spontaneous resting state. The time-compressed threshold and speech recognition threshold data indicated significant differences among the groups. In patients with age-related hearing loss, information masking (babble noise) had a greater effect than energy masking (speech-shaped noise) on processing difficulties. In terms of resting-state electroencephalography signals, we observed enhanced frontal lobe (Brodmann's area, BA11) activation in the older adults with normal hearing compared with the younger participants with normal hearing, and greater activation in the parietal (BA7) and occipital (BA19) lobes in the individuals with age-related hearing loss compared with the younger adults. Our functional connection analysis suggested that compared with younger people, the older adults with normal hearing exhibited enhanced connections among networks, including the default mode network, sensorimotor network, cingulo-opercular network, occipital network, and frontoparietal network. These results suggest that both normal aging and the development of age-related hearing loss have a negative effect on advanced auditory processing capabilities and that hearing loss accelerates the decline in speech comprehension, especially in speech competition situations. Older adults with normal hearing may have increased compensatory attentional resource recruitment represented by the top-down active listening mechanism, while those with age-related hearing loss exhibit decompensation of network connections involving multisensory integration.

Development and characterization of BAC-end sequence derived SSRs, and their incorporation into a new higher density genetic map for cultivated peanut (Arachis hypogaea L.).

BACKGROUND: Cultivated peanut (Arachis hypogaea L.) is an important crop worldwide, valued for its edible oil and digestible protein. It has a very narrow genetic base that may well derive from a relatively recent single polyploidization event. Accordingly molecular markers have low levels of polymorphism and the number of polymorphic molecular markers available for cultivated peanut is still limiting. RESULTS: Here, we report a large set of BAC-end sequences (BES), use them for developing SSR (BES-SSR) markers, and apply them in genetic linkage mapping. The majority of BESs had no detectable homology to known genes (49.5%) followed by sequences with similarity to known genes (44.3%), and miscellaneous sequences (6.2%) such as transposable element, retroelement, and organelle sequences. A total of 1,424 SSRs were identified from 36,435 BESs. Among these identified SSRs, dinucleotide (47.4%) and trinucleotide (37.1%) SSRs were predominant. The new set of 1,152 SSRs as well as about 4,000 published or unpublished SSRs were screened against two parents of a mapping population, generating 385 polymorphic loci. A genetic linkage map was constructed, consisting of 318 loci onto 21 linkage groups and covering a total of 1,674.4 cM, with an average distance of 5.3 cM between adjacent loci. Two markers related to resistance gene homologs (RGH) were mapped to two different groups, thus anchoring 1 RGH-BAC contig and 1 singleton. CONCLUSIONS: The SSRs mined from BESs will be of use in further molecular analysis of the peanut genome, providing a novel set of markers, genetically anchoring BAC clones, and incorporating gene sequences into a linkage map. This will aid in the identification of markers linked to genes of interest and map-based cloning.

Estimation of leak area-pressure relationship for cracks on water pipes using models based on linear-elastic fracture mechanics.

The assessment and control of the real losses from water distribution systems require the accurate estimation of the flow rate from an individual leak as a function of the internal pressure. The lack of analytical models able to accurately describe the relationship between the area of the leak and the pressure head is the key problem. This paper utilized the linear-elastic fracture mechanics (LEFM) theory for thin shells to derive models for both longitudinal and circumferential cracks. The models were validated by both finite element (FE) simulations and laboratory experiments under varying crack and pipe parameters. Both fluid-structure interaction (FSI) and traditional FE simulations were performed, and the results were compared to quantify the effect of leakage hydraulics on leak area. In the laboratory experiments, an image analysis technology was utilized to measure the leak area and flow rate simultaneously, so that the effect of the discharge coefficient could be excluded. In addition, the leak area was systematically measured under the effect of different parameters. The results revealed that the values predicted by the derived models were in good agreement with the experimental and FE simulation values for both types of cracks. The LEFM theory and the phenomena observed in this study can improve our understanding of the leak behavior and enable the development of effective pressure management strategies for water distribution systems.

Secondary coronary artery ostial lesions: Three case reports.

BACKGROUND: Atherosclerosis is one of the main causes of coronary artery ostial lesions seen clinically. Secondary coronary artery ostial lesions are rare, and cases reported previously were associated with syphilitic vasculitis and aortic dissection. Here, we report three rare cases of secondary coronary ostial lesions. Due to their rareness, these lesions can easily be neglected, which may lead to misdiagnosis and missed diagnosis. CASE SUMMARY: We present three patients with acute myocardial infarction and unstable angina caused by secondary coronary artery ostial lesions. In Case 1, coronary angiography (CAG) revealed 90% stenosis of the left main coronary ostium. Chest contrast computed tomography (CT) suggested thymic carcinoma invading the left main coronary ostium. Coronary artery bypass grafting and tumor resection were performed. In Case 2, echocardiography revealed a sinus of Valsalva aneurysm (SVA)-like dilatation. CAG showed a right coronary sinus giant aneurysm and complete obstruction of the right coronary artery (RCA) ostium. Aortic contrast CT confirmed these findings. The Bentall procedure was performed. In Case 3, CT CAG identified an anomalous origin of the right coronary artery (AORCA) from the left sinus of Valsalva coursing between the aorta and pulmonary trunk, causing severe RCA ostium stenosis by compression. Surgical correction of the AORCA was performed. CONCLUSION: The cases reported here suggest that we should consider other causes of coronary ostial lesions other than atherosclerosis.

A Deep Learning Approach to Predict Conductive Hearing Loss in Patients With Otitis Media With Effusion Using Otoscopic Images.

Importance: Otitis media with effusion (OME) is one of the most common causes of acquired conductive hearing loss (CHL). Persistent hearing loss is associated with poor childhood speech and language development and other adverse consequence. However, to obtain accurate and reliable hearing thresholds largely requires a high degree of cooperation from the patients. Objective: To predict CHL from otoscopic images using deep learning (DL) techniques and a logistic regression model based on tympanic membrane features. Design, Setting, and Participants: A retrospective diagnostic/prognostic study was conducted using 2790 otoscopic images obtained from multiple centers between January 2015 and November 2020. Participants were aged between 4 and 89 years. Of 1239 participants, there were 209 ears from children and adolescents (aged 4-18 years [16.87%]), 804 ears from adults (aged 18-60 years [64.89%]), and 226 ears from older people (aged >60 years, [18.24%]). Overall, 679 ears (54.8%) were from men. The 2790 otoscopic images were randomly assigned into a training set (2232 [80%]), and validation set (558 [20%]). The DL model was developed to predict an average air-bone gap greater than 10 dB. A logistic regression model was also developed based on otoscopic features. Main Outcomes and Measures: The performance of the DL model in predicting CHL was measured using the area under the receiver operating curve (AUC), accuracy, and F1 score (a measure of the quality of a classifier, which is the harmonic mean of precision and recall; a higher F1 score means better performance). In addition, these evaluation parameters were compared to results obtained from the logistic regression model and predictions made by three otologists. Results: The performance of the DL model in predicting CHL showed the AUC of 0.74, accuracy of 81%, and F1 score of 0.89. This was better than the results from the logistic regression model (ie, AUC of 0.60, accuracy of 76%, and F1 score of 0.82), and much improved on the performance of the 3 otologists; accuracy of 16%, 30%, 39%, and F1 scores of 0.09, 0.18, and 0.25, respectively. Furthermore, the DL model took 2.5 seconds to predict from 205 otoscopic images, whereas the 3 otologists spent 633 seconds, 645 seconds, and 692 seconds, respectively. Conclusions and Relevance: The model in this diagnostic/prognostic study provided greater accuracy in prediction of CHL in ears with OME than those obtained from the logistic regression model and otologists. This indicates great potential for the use of artificial intelligence tools to facilitate CHL evaluation when CHL is unable to be measured.

Gene interfered-ferroptosis therapy for cancers.

Although some effective therapies have been available for cancer, it still poses a great threat to human health and life due to its drug resistance and low response in patients. Here, we develop a ferroptosis-based therapy by combining iron nanoparticles and cancer-specific gene interference. The expression of two iron metabolic genes (FPN and LCN2) was selectively knocked down in cancer cells by Cas13a or microRNA controlled by a NF-κB-specific promoter. Cells were simultaneously treated by iron nanoparticles. As a result, a significant ferroptosis was induced in a wide variety of cancer cells. However, the same treatment had little effect on normal cells. By transferring genes with adeno-associated virus and iron nanoparticles, the significant tumor growth inhibition and durable cure were obtained in mice with the therapy. In this work, we thus show a cancer therapy based on gene interference-enhanced ferroptosis.

Erratum: A New Tool for CRISPR-Cas13a-BasedCancer Gene Therapy.

[This corrects the article DOI: 10.1016/j.omto.2020.09.004.].

A One-Pot CRISPR/Cas9-Typing PCR for DNA Detection and Genotyping.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated endonuclease Cas9 (Cas9) has high specificity to its target DNA as a gene editing tool. This characteristic makes it useful for DNA detection. Combining the advantages of CRISPR/Cas9 and PCR, this study establishes a novel CRISPR/Cas9-based DNA detection method, named CRISPR/Cas9-typing PCR version 4.0 (ctPCR4.0). This method can detect target DNA in one pot with high specificity and sensitivity. In a homogenous reaction, the target DNA is first cleaved by a pair of Cas9- single-guide RNA complexes and thus releases two single strands with free 3' ends, allowing a pair of oligonucleotides to anneal with the strands. The annealed oligonucleotides provide templates for DNA polymerization from the free 3' ends. A universal primer annealing site is thus produced at the end of two single strands. The target DNA is then amplified by PCR using a universal primer. This method was first verified by accurately detecting the cloned L1 fragments of 10 genotypes of high-risk human papilloma viruses (HPVs). This method was then validated by detecting the L1 fragments of two highest-risk HPVs, HPV 16 and HPV 18, in the genomic DNA of two HPV-positive cervical carcinoma cells, HeLa and SiHa. Finally, this method was further validated by accurately detecting 10 high-risk HPVs in 30 clinical samples.

Development of BRAF V600E Mutation in NRAS Q61L Mutated Rectal Cancer.

BRAF and NRAS are oncogenes in the RAS/RAF/MEK/MAP-kinase signaling pathway. Coexistent mutations of BRAF and NRAS in a single colorectal cancer patient have always been considered mutually exclusive or at least rare. The clinical outcome of these patients remains undetermined. Herein we report a 53-year-old man harboring an NRAS Q61L mutation in his primary rectal carcinoma, who presented with a concomitant mutation of BRAF V600E in his liver metastasis biopsy 55 months after the primary CRC surgical resection. Our findings suggest that a BRAF and NRAS developed co-mutation may lead to a distinct clinicopathological progression. BRAF-mutated CRCwill not benefit from anti-RAS targeted therapy.

Detecting and differentiating Theileria sergenti and Theileria sinensis in cattle and yaks by PCR based on major piroplasm surface protein (MPSP).

Theileria sergenti and Theileria sinensis are closely related members of benign Theileria species found in cattle and yaks in China. They are morphologically indistinguishable. A polymerase chain reaction (PCR) targeting major piroplasm surface protein of T. sergenti and T. sinensis was developed in this study. The newly developed oligonucleotide primer set was able to specifically amplify the DNA of T. sinensis and in conjunction with primers for T. sergenti and these two species could be detected and distinguished. Specificity testing also revealed that there was no cross-reaction with the other tick-borne diseases Theileria annulata, Babesia ovata, Anaplasma marginale as well as bovine white blood cells. Phylogenetic analysis based on the MPSP gene sequences confirmed the specificity of PCR assays. The sensitivity of the methods was 0.1pg DNA for the T. sergenti PCR and 1pg DNA for T. sinensis PCR. Two hundred and thirty-six field blood samples from of cattle and yaks were collected from five different geographical regions in China where benign Theileria species have been found. T. sergenti was found in all five provinces but was absent from one county in Gansu Province. T. sinensis was only found in Gansu Province. In both counties in Gansu where the parasites co-existed, mixed infections were detected. Our results indicate that the PCR methods developed in this study are suitable for the detection and differentiation of T. sergenti and T. sinensis.

Sequence analysis of Theileria annulata surface protein in Chinese isolates.

OBJECTIVE: To study the TaSP polymorphism in three Chinese isolates of Theileria annulata. METHODS: The isolates from Inner Mongolia Autonomous Region, Ningxia Hui Autonomous Region and Xinjiang Uygur Autonomous Region were cultured in RPMI 1640 medium. TaSP gene was amplified from genomic DNA extracted from schizonts using polymerase chain reaction (PCR) and sequenced. Its amino acid sequence comparison was carried out with Clustal W2 multiple sequence alignment program. Molecular component and motif prediction were performed with online servers. RESULTS: The comparison of TaSP amino acid sequences of the three isolates showed that the central region (aa position 38-161) predicted to be the highly immunogenetic domain was polymorphic both in size and amino acid sequence, while the N-terminal (first 37 aa) and C-terminal (last 154 aa) parts were strongly conserved. Phylogenetic analysis and percentage identity revealed that the Chinese isolates were closely related to the isolates from Turkey, but quite different from those of India, Morocco and Tunisia. More importantly, variability was noticed among Chinese isolates, which caused both the location and number's differences of motif (casein kinase II phosphorylation sites) among three TaSP sequences. CONCLUSION: TaSP polymorphism exists in the Chinese isolates of T. annulata.