Sterility of Aedes albopictus by X-ray Irradiation as an Alternative to γ-ray Irradiation for the Sterile Insect Technique.

The mosquito Aedes albopictus can transmit various arboviral diseases, posing a severe threat to human health. As an environmentally friendly method, sterile insect technology (SIT) is considered an alternative to traditional methods such as chemical pesticides to control Ae. albopictus. In SIT, the sterility of male mosquitoes can be achieved by γ-ray or X-ray radiation. Compared with γ-rays, X-rays are easier to obtain, cheaper, and less harmful. However, there is a lack of comparative assessment of these two types of radiation for SIT under the same controlled conditions. Here, we compared the effects of X-ray and γ-ray radiation on the sterility of Ae. albopictus males under laboratory-controlled conditions. Neither type of radiation affected the number of eggs but significantly reduced the survival time and hatch rate. The same dose of γ-rays caused a higher sterility effect on males than X-rays but had a more significant impact on survival. However, X-rays could achieve the same sterility effect as γ-rays by increasing the radiation dose. For example, X-rays of 60 Gy induced 99% sterility, similar to γ-rays of 40 Gy. In the test of male mating competitiveness, the induced sterility and the male mating competitiveness index were also identical at the same release ratio (sterile males/fertile males). At a release ratio of 7:1, nearly 80% of eggs failed to hatch. Sterile males produced by X-ray and γ-ray radiation had similar male competitiveness in competition with field males. In conclusion, a higher dose of X-rays is required to achieve the same sterility effect, compared to γ-rays. When γ-rays are not readily available, high-dose X-rays can be used instead. This study provides data supporting the selection of more suitable radiation for the field release of sterile male mosquitoes.

An Efficient Schistosoma japonicum Bivalent Membrane Protein Antigen DNA Vaccine Against Schistosomiasis in Mice.

BACKGROUND Schistosomiasis is one of the most important infectious parasitic diseases in the world. The most important was to control schistosomiasis is through a combination of medical therapy and immunization. The membrane antigens Tsp2 and 29 from Schistosoma are promising anti-schistosomiasis vaccine candidates. MATERIAL AND METHODS In this study, the pcDNA3.1(+)-SjTsp2, pcDNA3.1(+)-Sj29, and pcDNA3.1 (+)-SjTsp2-29 eukaryotic expression vectors were successfully constructed as DNA vaccines, and the protective abilities of these vaccines were evaluated in mice. RESULTS The results showed that vaccination with SjTsp2, Sj29, and SjTsp2-29 reduced parasite burden and hepatic pathology compared to the control group, and the protective effect of the bivalent SjTsp2-29 DNA vaccine was better than that of the univalent SjTsp2 or Sj29 DNA vaccines. We also found high levels of IgG, IgG1, and IgG2a against SjTsp2, Sj29, and SjTsp2-29 DNA vaccines, with high expression of IFN-γ and no IL-4 in the mice. CONCLUSIONS The double-membrane antigen DNA vaccine SjTsp2-29 elicited protection against Schistosoma infection and might serve as a vaccine candidate.

Potential role of IL-37 signaling pathway in feedback regulation of autoimmune Hashimoto thyroiditis.

IL-37, the anti-inflammatory cytokine of the IL-1 family, plays several key roles in the regulation of autoimmune diseases. Yet, its role in Hashimoto's thyroiditis (HT) is not clear. In the present study, we found that, in tissues from HT patients, most of the follicular epithelial cells were positive for both IL-37 and single Ig IL-1-related receptor (SIGIRR) by immunohistochemical staining, while the infiltrating lymphocytes and other inflammatory cells hardly expressed any. Meanwhile, mRNA expression levels of IL-37 in peripheral blood mononuclear cells (PBMC) of HT patients were significantly higher than those in normal controls measured by quantitative real-time PCR. Finally, we studied the possible role of IL-37 in IFN-γ-stimulated rat FRTL-5 cells. The results showed that IL-1ß, TNF-α, and MCP-1 mRNA levels were significantly decreased, while the expression of IL-4 mRNA was dramatically up-regulated in IFN-γ-stimulated rat thyroid cell line FRTL-5 pre-treated with IL-37. The current study, for the first time, demonstrated that the IL-37 network is involved in Hashimoto's thyroiditis, and IL-37 signaling pathway may ameliorate the excessive autoimmune responses in this chronic lymphocytic thyroiditis.

Value of Apparent Diffusion Coefficient (ADC) and Dynamic Contrast-Enhanced Magnetic Resonance Imaging (DCE-MRI) in Differentially Diagnosing Angiomatous Meningiomas and Solitary Fibrous Tumors/Hemangiopericytomas.

BACKGROUND To determine if ADC and DCE-MRI can be used to differentiate angiomatous meningiomas (AMs) from solitary fibrous tumors/hemangiopericytomas (SFT/HPCs). MATERIAL AND METHODS We retrospectively reviewed records of 103 patients from 1 January 1 2014 to 1 November 2018. We enrolled 41 patients who had undergone a 3T MRI, with histologically confirmed AMs in 20 (48.80%) patients, and SFT/HPCs in 21 (51.20%) patients. The ADC and DCE-MRI features were derived and then compared by 2 independent-samples t tests and Wilcoxon rank sum test to obtain the ROC. RESULTS AMs had significantly lower ADC values than did SFT/HPCs, but AMs had significantly higher MCER values than did SFT/HPCs. A threshold value of 1.03×10⁻³ mm²/s for ADC to predict AMs from SFT/HPCs was estimated (AUC=0.902, sensitivity=88.20%, specificity=83.30%). Optimal diagnostic performance (AUC=0.825, sensitivity=84.60%, specificity=81.80%) was obtained when setting MCER=226.7% as the threshold value. CONCLUSIONS The ADC values of AMs were lower than those of SFT/HPCs; the MCER of AMs were greater than those of SFT/HPCs, and ADC was more useful than MCER, and these parameters could help diagnosis.

Histogram Analysis Parameters ADC for Distinguishing Ventricular Neoplasms of Ependymoma, Choroid Plexus Papilloma, and Central Neurocytoma.

BACKGROUND To determine if histograms of ADC can be used to differentiate ventricular ependymomas, choroid plexus papillomas (CPPs), and central neurocytomas (CNCs). MATERIAL AND METHODS We retrospectively reviewed records from 185 patients from 1 January 2014 to 1 November 2018. We finally included a total of 60 patients: 36 (60.00%) had histologically confirmed ependymomas, 10 (16.67%) had CPPs, and 14 (23.33%) had CNCs, as determined by routine MRI scanning at 3.0T. The ADC histogram features were derived and then compared by Kruskal-Wallis test (they were not normally distributed). Bonferroni test was used to compare the 2 groups and then we determined the ROC. RESULTS Ependymomas had significantly higher mean, perc.01%, perc.10%, perc.50%, perc.90%, and perc.99% than CNCs. Ependymomas had significantly lower skewness than CNCs. Histogram metrics derived from mean, perc.01%, perc.10%, perc.50%, and perc.90% were significantly lower in the CNCs group than in the CPPs group. CPPs showed significantly lower skewness than CNCs. A threshold value of 86.50 for perc.50% to predict ependymomas from CNCs was estimated (AUC=0.97, sensitivity=97.20%, specificity=85.70%). Optimal diagnostic performance to predict CPPs from CNCs (AUC=0.96, sensitivity=100.00%, specificity=85.70%) was obtained when setting Perc.50%=84.00 as the threshold value. CONCLUSIONS The ADC histogram analysis may help to discriminate ependymomas, CPPs, and CNCs.

Development of monoclonal antibodies against Sj29 and its possible application for schistosomiasis diagnosis.

OBJECTIVE: Timely Schistosoma japonicum detection improves outcomes in schistosomiasis. Here, we established a double antibody sandwich ELISA to detect Schistosoma japonicum. METHODS: Sj29 polyclonal and monoclonal antibodies were developed and identified. A Sj29 double antibody sandwich ELISA was evaluated. RESULTS: Assay sensitivity for detecting Schistosoma japonicum circulating antigen Sj29 was 76.7% (23/30), 54.5% (18/33) and 50.0% (18/36) in patients with acute, chronic and advanced schistosomiasis. No false positives or cross-reactivity was observed in healthy controls or patients with clonorchiasis, paragonimiasis, or ancylostomiasis, respectively. By contrast, false positives (5.7%) and cross-reactivity (6.5%-10%) were detected using an AWA-ELISA. The circulating antigen positive rates decreased significantly faster than that of the antibody detection after 6 months treatment (22.2%, 4/18 and 88.9%, 16/18). Chi-Square Tests revealed that Sj29 sandwich ELISA had lower sensitivity than AWA indirect ELISA in the detection of S. japonicum infected patients (p<0.05). Although our assay detection specificity in patients infected with other parasites or healthy controls appeared higher, the difference between the assays was insignificant. However, our assay showed significantly better results in monitoring praziquantel therapeutic effects (p=0.001), with antigen-positive rates decreasing significantly faster than antibody detection rates after 6 months of treatment (22.2%, 4/18 versus 88.9%, 16/18). CONCLUSIONS: Sj29 double antibody sandwich ELISA was established. The specificity of this method for detecting healthy sera was 100%. Meanwhile, Sj29 sandwich ELISA may have a potential diagnostic capability to distinguish current from past infections and assess drug treatment responses.

Conventional MRI for diagnosis of subacute combined degeneration (SCD) of the spinal cord due to vitamin B-12 deficiency.

Subacute combined degeneration of the spinal cord (SCD) is often found in vitamin B-12 deficiency and typically shows hyperintensity on T2-weighted images of the lateral and posterior columns. The purpose of the study was to evaluate the use of conventional magnetic resonance examination in diagnosing SCD. Thirty-six patients were clinically confirmed and retrospectively analyzed; conventional spine MRIs were available for all patients and eight of them had contrast enhancement MRIs. 19 out of 36 patients showed abnormal signal intensity on T2 weighted images with a sensitivity of 52.8%, among which 18 in the posterior aspect of the spinal cord and 1 in the anterior horn of the thoracic spinal cord The spinal cord abnormalities were seen at the cervical spine in 12 patients (33.3%) and at the thoracic spine in the other 7 patients (19.4%). Axial T2-weighted images showed symmetric linear T2-hyperintensity as an "inverted V" at the cervical spinal cord in 5 patients, which has been reported as a typical sign of SCD. For patients with thoracic spinal cord abnormalities, the bilateral paired nodular T2-hyperintensity looked like "binoculars" at the thoracic spinal cord. Only one out of the eight patients showed slight enhancement after injection with contrast agent. All the 36 patients reported clinical improvement after appropriate vitamin B-12 treatment. The two follow-up spine MRIs showed a decreased extent of the lesion. Therefore, conventional MRI is useful in the diagnosis and management of SCD caused by vitamin B-12 deficiency.

Schistosoma japonicum: Tsunagi/Y14 protein plays a critical role in the development of the reproductive organs and eggs.

Tsunagi/Y14 is an evolutionarily conserved RNA-binding protein that is required for the maintenance of oogenesis and the masculinization of the germ-line in many animal models. We speculated that Tsunagi/Y14 might also regulate reproductive organ development in Schistosoma japonicum (S. japonicum, Sj). Sj Tsunagi/Y14 and control double-stranded RNAs were introduced into schistosomula by electroporation respectively. These transfected schistosomula were cultured in vitro for 1, 3 or 5 days. The mRNA and protein levels of the target gene in the cultured schistosomula were significantly suppressed compared with those of the control group. Furthermore, BALB/c mice were infected with the transfected schistosomula for 6 weeks and were sacrificed to harvest the adult worms. We found that the silencing of Sj Tsunagi/Y14 led to defects in reproductive organs development in both male and female worms. Moreover, it also affected the size, quantity and activity of the eggs in the mice liver. Our findings indicated that Tsunagi/Y14 plays a critical role in the development of reproductive organs and eggs in S. japonicum.

Worm morphology of Schistosoma japonicum using confocal laser scanning microscopy.

Male and female Schistosoma japonicum worms have dissimilar appearances in their final host. In this study, a morphometric and morphological assessment of whole worms derived from unisexual and mixed infections in mice was conducted using confocal laser scanning microscopy. Worms from mixed infections showed significant morphological changes between 15 and 25 days post-infection (PI). On the fifteenth day PI, 33% of males had formed the conspicuous gynecophoric canal, but only 8% of them had testicular lobes containing a few germinative cells; 13% of females had incipient ovaries with a few immature ovarian cells inside. On the twentieth day PI, the testicular lobes contained more germinative cells in all male worms, while female worms presented vitelline glands. On the twenty-fifth day PI, more germinative cells were observed in the male testicular lobes, and differentiated cells were present in the female ovaries. All worms had fully developed reproductive organs from 30 days PI onwards. Morphometric analysis showed significant differences between mixed and unisexual infections at 35 days PI. Ovaries of worms from unisexual infections contained cells in one stage of maturation and vitelline glands had undifferentiated cells. Our study of S. japonicum provides a detailed comparison of different morphological traits from worms of mixed and unisexual infections throughout development.

[Preliminary function study on Mago nashi gene of Schistosoma japonicum].

OBJECTIVE: To study the function of Mago nashi gene in reproductive system of Schistosoma japonicum. METHODS: dsRNA products of SjMago nashi gene and control gene (lacZ) were generated by in vitro transcription. SjMago nashi dsRNA and control (lacZ) dsRNA were electroporated into mechanically transformed schistosomula. Aliquots of parasites (1000) were harvested at day 1, 3, and 5 after electroporation, respectively. Total RNA and proteins were isolated simultaneously using TRIzol reagent. Levels of SjMago nashi mRNA and protein were determined by RT-PCR and Western blotting analysis, respectively. About 1000 dsRNA-electroporated schistosomula were injected into each BALB/c female mouse. Six weeks later the worms were collected, fixed, stained, clarified, dehydrated and mounted. The male and female reproductive organs were observed and measured under the confocal laser scanning microscope. RESULTS: At day 1, 3 and 5 post-electroporation, 22%, 69%, and 80% reduction in Mago nashi mRNA levels were detected respectively in SjMago nashi dsRNA-electroporated schistosomula (experiment group) compared to parasites treated with control dsRNA (control group); and schistosomula of experiment group exhibited 12%, 39%, and 56% decreased in Mago nashi protein expression levels in comparison to the control group, respectively. In experiment group there were many spermatozoa in testicular lobes and no changes were observed in ovary and vitelline gland. Compared to control group, adult worms in experiment group were smaller in the body width, the width and length of testicles and ovaries (P < 0.05). CONCLUSION: Mago nashi dsRNA can specifically inhibit the expression of target gene and protein. SjMago nashi gene is a reproduction-related gene.

[Evaluation of recombinant 29,000 extra membranous protein for the immunodiagnosis of schistosomiasis japonica].

OBJECTIVE: To evaluate the recombinant membranous protein 29000 (rSj29) of Schistosoma japonicum in the immunodiagnosis of schistosomiasis by indirect ELISA. METHODS: 394 serum samples collected from two low-endemic villages in Anhui Province were tested by rSj29-ELISA and AWA-ELISA using adult worm antigen (AWA) of S. japonicum. Meanwhile all subjects were each tested by 9 examinations with 3 fecal samples by modified Kato-Katz method, and by indirect hemagglutination assay (IHA). RESULTS: It was found that the positive detection rates of stool examination, IHA, rSj29-ELISA and AWA-ELISA were 4.8% (19/394), 62.2% (245/394), 68.3% (269/394), and 89.9% (354/394), respectively. There was no statistical difference between rSj29-ELISA and IHA (chi2 = 3.2, P > 0.05) with the coincidence of 80.7% (318/394). The coincidence of IHA, rSj29-ELISA and AWA-ELISA with stool examination was 42.6% (168/394), 36.0% (142/394), and 15.0% (59/394), respectively. CONCLUSION: The indirect ELISA with rSj29 antigen shows similar effect as IHA for the diagnosis of schistosomiasis.

[Construction of DNA vaccine pcDNA3.1(+)/tetraspanin 2-A against Schistosoma japonicum and its immune-protective effect in mice].

Tetraspanin 2-A (SjTsp2-A) gene was amplified by PCR. pcDNA3.1(+)/SjTsp2-A recombinant plasmids were constructed and transformed into E. coli DH5alpha. Twenty four BALB/c mice were randomly divided into pcDNA3.1(+)/ -SjTsp2-A group (A), pcDNA3.1(+)/SjGST group (B) and pcDNA3.1(+) group (C). Each mouse was injected through musculus quadriceps femoris by three times (two weeks interval) respectively with 100 microg pcDNA3.1(+)/SjTsp2-A, pcDNA3.1 (+)/SjGST, or pcDNA3.1(+). At two weeks after the final inoculation, mice were each challenged by 40 +/- 2 cercariae of S. japonicum. Forty-five days after infection, all mice were sacrificed, the number of worms collected and eggs in liver tissue was counted. Anti-pcDNA3.1(+)/SjTsp2-A antibody was detected by ELISA and protein expression in quadriceps muscle by immunohistochemical staining. The worm reduction rate (44.4%) and egg reduction rate (28.4%) of group A was higher than those of group B and C (P < 0.05), but no significant difference between groups B (3.9%, 19.3%) and C. Higher antibody titer (1:25 600) was detected in sera of group A. Immunohistochemistry analysis showed an expression of specific antigens in quadriceps muscles of groups A and B. The DNA candidate vaccine induces partial protective immunity against S japonicum in BALB/c mice.

[Cloning, expression and identification of membrane protein Tetraspanin 2-A of Schistosoma japonicum].

OBJECTIVE: To clone and express a membrane protein (Tetraspanin 2) gene of Schistosoma japonicum (SjTsp2). METHODS: A pair of primers was designed to amplify the SjTsp2 gene which was subcloned into prokaryotic plasmid pET28a(+). The recombinant plasmid was transformed into E. coli BL21(D3) and followed by expression of the protein induced by IPTG. The protein was purified by affinity chromatography and used to immunize BALB/c mice. Dilution of antibody against SjTsp2 was determined by ELISA. The protein was also identified by Western blotting. RESULTS: Big loop of SjTsp2-A, 228 bp, was amplified in vitro by PCR. Its deduced amino sequence shared 52% similarity with SmTsp2. The soluble recombinant SjTsp2-A was expressed in the experiment and high dilution antibody against the recombinant (1:32,000 in maximum) was produced in immunized mice. SjTsp2-A reacted positively with sera of acute and chronic schistosomiasis patients but not with sera from healthy persons by Western blotting. CONCLUSION: SjTsp2 has been expressed and shows certain antigenicity.

Schistosoma japonicum: inhibition of Mago nashi gene expression by shRNA-mediated RNA interference.

RNA interference (RNAi) mediated by short interfering RNA (siRNA) is a powerful reverse genetics tool and holds enormous therapeutic potential for various diseases, including parasite infections. siRNAs bind their complementary mRNA and lead to degradation of their specific mRNA targets. RNAi has been widely used for functional analysis of specific genes in various cells and organisms. In this paper, we tested the potential of silencing the expression of the Mago nashi gene in Schistosoma japonicum by siRNAs derived from shRNA expressed by mammalian Pol III promoter H1. Schistosomula, transformed from cercariae by mechanical shearing of the tails, were electroporated with Mago nashi shRNA expression vector. Aliquots of parasites were harvested at days 1, 3, and 5 after electroporation, respectively. Levels of Mago nashi mRNA and protein were determined by RT-PCR and Western blotting analysis. The results showed that shRNA expressed from mammalian Pol III promoter H1 specifically reduced the levels of Mago nashi mRNA and proteins in S. japonicum. Changes in testicular lobes were apparent when parasites were introduced into mammalian hosts. Thus, vector-mediated gene silencing is applicable to S. japonicum, which provides a means for the functional analysis of genes in this organism.

[Morphological observation on the reproductive system of adult Schistosoma japonicum under confocal laser scanning microscopy].

BALB/c mice were infected with Schistosoma japonicum cercariae (40+/-2 per mouse) through abdominal skin. Mice were sacrificed after 35 days to acquire the adult worms which were then fixed, stained, clarified, dehydrated and mounted. The specimens were observed under the confocal laser scanning microscope. The overall morphology of the adult worms was displayed distinctly, especially the testicular lobes, seminal vesicle and genital pore of the male, reproductive system, and the ovary, vitelline glands, oviduct, vitelline duct, seminal receptacle, ootype, mehlis gland, uterus, genital pore and eggs of female reproductive system. The confocal laser scanning microscopy is an alternative method to research organs, tissues and cell structure of schistosome worm.