[One case of myocardial damage caused by carbamate pesticide poisoning].

The data of a patient with carbamate pesticide poisoning were analyzed. Cardiac arrest, oliguria, acute renal injury and pulmonary infection occurred during treatment. After cardiopulmonary resuscitation, tracheal intubation, CRRT, anti-infection and other symptomatic support treatment, the patient recovered and discharged. The myocardial damage caused by carbamate pesticide poisoning is easy to be ignored, and it often causes cardiac manifestations such as arrhythmia and cardiac insufficiency, and the related markers of cardiac injury, electrocardiogram and echocardiogram are also changed. Therefore, the awareness of cardiac damage caused by carbamate pesticide poisoning should be improved.

[Establishment of morphological reference values for the differential count of white blood cells in peripheral blood smear, as well as nucleated cells and megakaryocytes in bone marrow smear].

Objective: To establish the morphological reference values for the differential count of white blood cells in peripheral blood smear as well as nucleated cells and megakaryocytes in bone marrow smear. Methods: From April 2012 to June 2020, 4 221 healthy donors for hematopoietic stem cell transplantation in Hebei Yanda Lu Daopei Hospital were selected. The median age was 36 (3-72) years old, including 2 520 males and 1 701 females. They were divided into four groups according to age: children group, with age≤14 years old [n=334, 11 (3-14) years old], youth group, with age >14 years old and <45 years old [n=2 855, 33 (15-44) years old], middle-aged adult group, with age ≥45 years old and < 60 years old [n=929, 49 (45-59) years old], and older adult group, with age ≥60 years old [n=103, 62 (60-72) years old]. Gender subgroups were established in each age group. According to different hematopoietic characteristics, the children group were divided into two subgroups: children group 1 [n=48, 6 (3-7) years old] and children group 2 [n=286, 11 (8-14) years old]. According to the clinical routine, 100 white blood cells in peripheral blood, 200 nucleated cells in bone marrow, and cell numbers/4.5 cm2 for megakaryocytes were classified and counted. The results of cell count in different age and gender groups were compared, and the reference values of morphological classification were established for different groups with statistical or clinical significance. Results: Due to the existence of statistically significant differences between children and adult groups and different gender subgroups in adults (all P<0.05), the reference values were established for children group and adult gender subgroups. The counts of segmented neutrophils and lymphocytes in peripheral blood were 46.65(43.97-49.32)% and 44.00(10.60-65.10)% in children group 1, 50.73(49.50-51.96)% and 39.55 (38.36-40.74)% in children group 2, and 57.00 (39.00-75.23) % and 33.00 (17.00-52.00) % in adult group, respectively. Bone marrow segmented neutrophils, orthochromatic erythroblasts, and mature lymphocytes were 11.54 (10.68-12.41)%, 14.20 (13.19-15.21)%, and 23.99 (22.06-25.92)% in children group 1, 12.50 (7.00-21.50)%, 15.00(9.50-25.50)%, and 21.02 (20.24-21.81)% in children group 2, 13.50 (7.50-21.00)%, 16.50 (10.50-26.00)%, and 15.50 (7.50-26.00)% in adult male group, and 14.50 (8.00-24.50)%, 14.50 (9.00-23.00)%, and 17.50 (8.50-29.00)% in adult female group, respectively. The myelopoiesis/erythropoiesis ratio in children group, adult male group and adult female group was 1.86∶1 (1.14∶1-3.23∶1), 1.96∶1 (1.12∶1-3.19∶1), 2.22∶1 (1.30∶1-3.69∶1), respectively. The numbers of granular megakaryocytes and thromocytogenic megakaryocytes were 138 (25-567) cells/4.5cm2 and 86 (13-328) cells/4.5 cm2 in children group, and 92 (13-338) cells/4.5 cm2 and 38 (3-162) cells/4.5 cm2 in adult group, respectively. Conclusion: The morphological reference values for the differential count of white blood cells in peripheral blood smear as well as nucleated cells and megakaryocytes in bone marrow smear are successfully established, which is helpful to improve the application of morphological examination in disease screening, diagnosis and monitoring.

[Research progress on the treatment of presbyopia].

Presbyopia is a physiological aging situation that the plasticity and elasticity of the lens and the function of the ciliary muscle become weaker, resulting in a decreased accommodation and inability to focus on near objects. Nowadays, there are many clinical strategies to correct presbyopia, each of which has its own advantages and disadvantages, however, there is no true sense of way to restore accommodation function. This article reviews both worldwide and domestic research on presbyopia, and analyzes and summaries the status quo as well as research progress of presbyopia correction modalities, surgical approaches, and drug therapies, hoping to provide a reference for clinical works.

A rapid sap-direct reverse transcription-polymerase chain reaction method for detection of dendrobium viroid in Dendrobium plants.

Dendrobium viroid (DVd) was first reported in China in 2020, and it is the only viroid known to infect Orchidaceae family plants. In this study, we developed a simple reverse transcription-polymerase chain reaction (RT-PCR) method for the rapid detection of DVd in Dendrobium plants. When extracting the sap template from the leaves, they are first clamped between two layers of plastic film, and the sap is pressed out and collected with a pipette. Using this sap, DVd was detected by dot-blot and RT-PCR methods and, the expected amplicons were confirmed by sequencing analysis. The batch analysis of field samples revealed that this method can be used to detect DVd rapidly. The detection method also reduces cross-contamination between different samples and minimizes false positives. Thus, this sap-direct RT-PCR method allows effective and rapid DVd detection in the study of Orchidaceae plants.

[Transcranial sonographic characteristics of Parkinson's disease with symptoms of restless legs syndrome].

Objective: To evaluate the transcranial sonographic characteristics in patients with Parkinson's disease (PD) with symptoms of restless legs syndrome (RLS). Methods: Patients with diagnosis of definite PD from the Second Affiliated Hospital of Soochow University and 3 other participating hospitals between September 2018 and December 2019 were consecutively enrolled. Concurrent RLS symptoms were determined using Non-motor Symptoms Questionnaire. Transcranial sonography (TCS) and clinical assessments were performed during the same time and the related variables were compared between the two groups using t-test, non-parametric test, Chi-square test and Spearman correlation analysis, respectively. Results: Among 349 patients with PD, the prevalence of RLS symptoms was 22.6%. Compared to patients without RLS symptoms, those with RLS had longer disease duration (43.0 (24.0, 91.0) months vs 37.0 (20.0, 60.0) months, P<0.05) and higher Hoehn-Yahr stage (2.5 (2.0, 3.0) vs 2.0 (1.5, 2.5), P<0.01).TCS revealed that patients with RLS symptoms were more likely to have abnormality in the raphe nucleus (21.50% vs 7.78%, χ²=15.9, P<0.001) and increased third ventricle width ((6.22±1.97) mm vs (5.16±1.90) mm, P<0.001). No significant differences were found regarding parameters of substantia nigra. Conclusions: Concurrent RLS symptoms are common in PD patients. Abnormal echogenicity of raphe nucleus and increased third ventricle width could be characteristics of TCS in PD patients with RLS symptoms.

[Cell morphological analysis of hepatosplenic T-cell lymphoma gamma-delta type].

Objective: To analyze the cell morphological features of hepatosplenic T-cell lymphoma (HSTCL) gamma-delta (γδ) type, differentiate from acute leukaemia (AL). Methods: This was a retrospective study. The clinical data of four cases of HSTCL γδ type who were treated in Hebei Yanda Ludaopei Hospital from 2009 to 2014 were collected. Their initial morphology diagnoses in other hospitals were all acute leukemia or myelodysplastic syndrome (MDS). Morphological analysis and cytochemical stains to their bone marrow (BM) aspiration and peripheral blood (PB) smears were completed when they had no response to previous chemotherapies, and the morphological reports were compared with results of immunophenotyping, chromosome, and T cell receptor (TCR) gene rearrangement. Results: The percentages of malignant cells in four patients' BM aspirations were 7.6%-40.0%, and in two patients' PB was 9% and 10%, respectively. The morphology of four cases had a very high similarity in Wright's stain. Predominantly medium-sized cells were seen, with rich cytoplasm and frequently one big conspicuous nucleolus. The malignant cells resembled blasts, especially monoblasts, but with coarse granular chromatin, more compact than that in monoblasts. When comparing to malignant myeloblast and lymphoblasts, HSTCL cells were larger and more irregular in cell shape, with more abundant cytoplasm and prominent nuclear irregularity. The cytochemistric stain played an important role in differential diagnosis. HSTCL malignant cells showed non-specific esterase (NSE) negative or focal punctate activity which couldn't be inhibited by sodium fluoride. Periodic acid-Schiff (PAS) stain was negative or positive with a form of coarse granules. The myeloperioxdase (MPO) stain was negative. Conclusion: Malignant cells of HSTCL γδ type have very distinct morphological features of mature lymphocytic neoplasm. The quality of Wright's stain, being short of complete cytochemical stains, lacking of awareness of this disease, and acute leukemia or MDS like appearance in some cases, result in the possibility of diagnostic error as malignant blast, and probably are main causes of misdiagnosis of HSTCL γδ type.

Identification of Dialeurodes citri as a Vector of Citrus yellow vein clearing virus in China.

In 2009, a new citrus viral disease caused by Citrus yellow vein clearing virus (CYVCV) was first discovered in China and now CYVCV is widely distributed in the field. CYVCV is transmissible by grafting and is spread by aphids from lemon to bean, and from bean to bean. However, until now, no vector has been shown to transmit CYVCV from citrus to citrus. In this study, after a 24-h acquisition access period (AAP), CYVCV was tested for in Dialeurodes citri (Ashmead), Panonychus citri McGregor, and Aphis citricidus (Kirkaldy) by quantitative RT-PCR. After an AAP of 48 h, groups of adults of D. citri, P. citri, and A. citricidus were given a 48 h inoculation access period on cultivar Daidai sour orange seedlings. Three, 6, and 12 months post-transmission by D. citri, CYVCV was detected in the receptor plants, and the mean incidence of infected trees was 31.9, 39.1, and 39.1%, respectively. CYVCV was not transmitted to citrus by P. citri or A. citricidus. This is the first report of the ability of D. citri to transmit CYVCV from infected to healthy citrus under laboratory conditions.

miR-483 is down-regulated in gastric cancer and suppresses cell proliferation, invasion and protein O-GlcNAcylation by targeting OGT.

MicroRNAs (miRNAs) are involved in the gastric carcinogenesis and progression. Here, we confirmed that miR-483 was frequently decreased in gastric cancer patients. The expression levels of miR-483 were negatively correlated with tumor stage, node metastasis and stromal invasion. Log-rank tests demonstrated that low expression of miR-483 was strongly correlated with poor overall survival in patients with gastric cancer. Moreover, ectopic expression of miR-483 remarkably suppressed gastric cancer cell proliferation by enhancing cell apoptosis and significantly inhibited the invasion of gastric cancer cells, while low expression of miR-483 exhibited the opposite effect. Bioinformatics analysis revealed that OGT was a potential target of miR-483, and miR-483 inhibited the expression level of OGT mRNA by direct binding to its 3'-untranslated region (3'UTR). Expression of miR-483 was negatively correlated with OGT in gastric cancer tissues. In addition, modulation of miR-483 expression could affect the global cellular protein O-GlcNAcylation in gastric cancer cells. Furthermore, silencing of OGT counteracted the effects of miR-483 repression, while its overexpression reversed tumor inhibitory effects of miR-483. In conclusion, our study revealed that miR-483 functions as a tumor suppressor by inhibiting proliferation, invasion and protein O-GlcNAcylation of gastric cancer via targeting OGT, and that miR-483 may serve as prognostic or therapeutic target for gastric cancer.

Occurrence, Distribution, and Molecular Characterization of Citrus yellow vein clearing virus in China.

In 2009, a new citrus viral disease caused by Citrus yellow vein clearing virus (CYVCV) was discovered in China. To more effectively monitor the presence of CYVCV, a survey was conducted in 166 citrus orchards from 11 major citrus-growing provinces in China from May 2014 to April 2016. In all, 458 of a total of 2,350 citrus samples tested positive for CYVCV, demonstrating that the virus is widely distributed in China. In this study, the complete genome sequences of 19 CYVCV isolates from different provinces and hosts were sequenced and characterized. Comparisons of the whole-genome sequences of these 19 CYVCV isolates as well as 4 isolates previously reported from around the world revealed that the sequence identity ranged from 97.1 to 99.8%, indicating that there is a very low level of sequence heterogeneity among CYVCV isolates of different geographic origins and hosts. Phylogenetic analysis of these 23 genomic sequences suggested that all of the isolates from China were clustered into the same clade, clearly apart from the CYVCV isolates from Turkey and Pakistan. To our knowledge, this is the first extensive survey conducted in China for CYVCV incidence.

Association of NF-κB1 gene polymorphisms with coronary artery disease in a Han Chinese population.

Nuclear factor (NF)-κB is a transcription factor that controls cell proliferation, differentiation, and immunity. Activated NF-κB1 is associated with the pathogenesis of coronary artery disease (CAD) and genetic polymorphisms in NF-κB1 have a plausible role in modulating the risk of CAD. To identify markers that contribute to the genetic susceptibility to CAD, we examined the potential association between CAD and single nucleotide polymorphisms (SNPs; rs28362491, rs230531, rs230528, rs1005819, rs4648055, rs3774964, and rs3774968) in the NF-κB1 gene using SNaPshot SNP genotyping assay. Participants included 361 patients with CAD and 385 healthy controls. The genotype and allele frequencies of the rs28362491 (promoter region) polymorphism in the CAD patients were significantly different from those in the healthy controls. The frequency of the D allele was significantly higher in CAD patients than in the healthy controls (P = 0.005 after Bonferroni correction). Strong linkage disequilibrium was observed in one block (D' > 0.9). Haplotype analysis revealed that haplotypes in block 1 of the NF-κB1 gene did not display a risk or protective effect (P > 0.05). These data suggest that NF-κB1 gene polymorphisms confer susceptibility to CAD and also support the notion that dysfunction of NF-κB1 is involved in the pathophysiological process of CAD.

Association between interleukin-17A polymorphism and coronary artery disease susceptibility in the Chinese Han population.

Coronary artery disease (CAD) is a major global health problem. In China, the incidence of CAD and the rate of mortality arising from it have increased every year. Interleukin-17A (IL-17A) is a proinflammatory cytokine produced by activated T cells, and it may be involved in the development of CAD. Genetic polymorphisms in functional regions of the IL17A gene have a plausible role in modulating the risk of CAD. To evaluate the role of IL17A polymorphisms as a risk factor for CAD, we performed a detailed analysis of possible functional single nucleotide polymorphisms (SNPs) in regulatory regions of IL17A. This study examined the potential association between CAD and five SNPs (rs8193037, rs8193036, rs3819024, rs2275913, and rs3748067) of the IL17A gene. The allelic or genotypic frequencies of the rs8193037 (promoter region) and rs8193036 (promoter region) polymorphisms in CAD were significantly different from those in healthy controls. The CAD subjects had a significantly lower frequency of the A allele of rs8193037 (P = 0.009, OR = 1.772, 95%CI = 1.146- 2.742) and the T allele of rs8193036 (P = 0.010, OR = 1.754, 95%CI = 1.139-2.701). Strong linkage disequilibrium was observed in one block (D' > 0.9). Significantly fewer T-G-G-A haplotypes (P = 0.045) were found in CAD subjects in block 1. These data suggest that IL17A gene polymorphisms confer susceptibility to CAD, and support the notion that dysfunction of IL-17A is involved in the pathophysiological process of CAD.

[Pregnancy outcomes and symptom improvement of patients with adenomyosis treated with high intensity focused ultrasound ablation].

Objective: To investigate the symptom improvement and pregnancy outcomes of patients with adenomyosis after treatment with high intensity focused ultrasound (HIFU) ablation. Methods: From October 2010 to October 2015, 68 patients with adenomyosis who wish to get pregnancies were treated with HIFU ablation in Suining Central Hospital. Among these patients, 56 presented with dysmenorrhea, 11 presented with menorrhagia, and 1 patient complained both; 41 of them had histories of abnormal pregnancy. The clinical data were analyzed retrospectively. Results: Fifty-four patients got pregnancy at the median of 10 months(range:1 to 31 months) after HIFU ablation, and 21 of them had delivered healthy babies. No uterine rupture occurred during gestation or delivery, and the newborn babies were healthy. Dysmenorrhea and menorrhagia in the patients who had pregnancies after HIFU ablation treatment were significantly relieved. The average menstruation volume score before and 1, 3, 6-month post-HIFU were 2.6±1.7, 1.7±0.8, 1.4±0.6, 1.3±0.6, respectively (P<0.05). The menstruation pain score before and 1, 3, 6-month after HIFU were 1.4±0.9, 0.9±0.7, 0.6±0.5, and 0.9±0.7, respectively (P<0.05). The volume of the adenomyotic lesions before and after HIFU at 1, 3, 6 month were (34±23), (23±15), (20±17), (20±12) cm3 (P<0.05). Although the spontaneous abortion rate was decreased after HIFU ablation treatment, there was no significant difference between the preoperative and postoperative [43% (23/54) versus 37% (20/54), P>0.05]. However, 20 of the 54 patients had spontaneous abortion, compared with 21 patients who had delivered babies, there were no significant statistical difference in terms of age, duration of disease, lesion size, non-perfused volume ratio, as well as the symptom scores before and after HIFU ablation treatment. Conclusions: HIFU ablation treatment is effective in improving symptoms of patients with adenomyosis. Based on our results, HIFU ablation treatment maybe also help to improve the pregnancy outcomes of patients with adenomyosis, but further studies are needed.

Comparison of high-intensity focused ultrasound therapy under nasal endoscopy guidance versus first-line drug treatment in patients with persistent allergic rhinitis.

The aim of this study was to determine whether high-intensity focused ultrasound (HIFU) therapy under nasal endoscopy guidance could provide better efficacy and safety in patients with persistent allergic rhinitis (PAR) than the first-line drugs recommended by the World Health Organization. A total of 120 adult patients with PAR were randomly divided into 2 groups (N = 60 each). One group underwent HIFU therapy under nasal endoscopy guidance using an ultrasound rhinitis therapeutic machine. The other group served as the control group and was treated with corticosteroid nasal spray and oral cetirizine hydrochloride. All patients underwent follow-up treatment for 1 year, after which the efficacy and safety were evaluated. There was no significant difference between the two groups (P > 0.05) in the total effective rate. Moreover, no complications such as nasal adhesion, septal perforation, mucosal atrophy, and hyposmia were observed, indicating that HIFU was as effective as the first-line drug treatments recommended by the World Health Organization for symptom relief in PAR patients. The treatment efficacy, repeatability, safety, economical aspects, ease of performance, and few complications of HIFU therapy strongly suggest that HIFU should be routinely incorporated into clinical practice.

Ultrastructural Changes and Putative Phage Particles Observed in Sweet Orange Leaves Infected with 'Candidatus Liberibacter asiaticus'.

Huanglongbing (HLB), also known as citrus greening, is currently the most destructive citrus disease. Anatomical analyses of HLB-affected sweet orange were carried out by light and electron microscopy. As compared with healthy citrus, the phloem plasmodesmata were plugged with callose, and in some samples the phloem was collapsed. Chloroplast structures were deformed. Prophage sequences occupy a significant portion of the genome of 'Candidatus Liberibacter asiaticus' and have been used to distinguish strains from Yunnan and Guangdong provinces in China and Florida. Interestingly, a large number of possible putative phage particles were observed attached on the surface of 'Ca. L. asiaticus' cells in plants inoculated with strain FJ3 from Fujian Province, China. Phage particles have been observed previously only in periwinkle plants artificially inoculated in Florida with 'Ca. L. asiaticus' that carried the SC1-type prophage. PCR assays verified the presence of the SC1-type prophage sequences previously described from this bacterium in Florida in the FJ3 isolate. This is the first time that suspected phage particles have been observed in sweet orange trees infected with 'Ca. L. asiaticus.'

Association of GDF5, SMAD3 and RUNX2 polymorphisms with temporomandibular joint osteoarthritis in female Han Chinese.

Temporomandibular joint osteoarthritis (TMJOA) is a complex disease and has a strong genetic component in its pathogenesis. Experimental evidence suggests the involvement of biological pathway in the disease. This case-control study was designed to investigate whether five common single nucleotide polymorphisms (SNPs) in GDF5, SMAD3, RUNX2, TGFß1 and CHST11, respectively, are associated with TMJOA in female Han Chinese patients. A total of 240 participants were evaluated comprising 114 female patients diagnosed with TMJOA based on Research Diagnostic Criteria for Temporomandibular Disorders and 126 healthy female controls. The SNPs of the five genes in the genomic DNA were examined by sequencing, and their allelic, genotypic and carriage rate frequency distributions, as well as the triple combination of the risk genotypes, were analysed using the logistic regression model. The SNP in GDF5 or SMAD3 showed significant association with TMJOA, a relatively weak association was observed in RUNX2. In the triple combinational analysis, the risk of TMJOA grew 5·09 times in the patients with five or six risk alleles (P < 0·01). This is the first study to evaluate the association of GDF5, SMAD3, RUNX2, TGFß1 and CHST11 with TMJOA in female Han Chinese. Our study suggests that the SNPs of genes related to TGFß family might contribute to the risk of TMJOA.

Percutaneous manual aspiration thrombectomy followed by stenting for iliac vein compression syndrome with secondary acute isolated iliofemoral deep vein thrombosis: a prospective study of single-session endovascular protocol.

OBJECTIVE: To evaluate the feasibility, safety, and effectiveness of single-session endovascular treatment with manual aspiration thrombectomy (MAT) as the first-line method of thrombus removal for iliac vein compression syndrome (IVCS) with secondary acute isolated iliofemoral deep vein thrombosis (DVT). METHODS: This was a prospective clinical study. Twenty-six patients (19 women, 7 men, mean age 54 years) with left-sided acute iliac-common femoral DVT secondary to IVCS were enrolled. All patients presented with leg swelling or pains. Endovascular treatment, consisting of MAT, balloon angioplasty, and stent placement, was performed in the same setting. Overnight antegrade thrombolysis was performed in patients with residual thrombus after MAT. Patients were followed up by ultrasonography. The mean follow-up period was 17.8 months (12-25 months). RESULTS: Single-session endovascular procedures were performed successfully in all patients. The mean procedure time was 67 minutes (ranging from 45 to 90 minutes). Complete thrombus removal, including almost 100% of removal in 24 patients and little residual thrombus (<5%) in two, was achieved after repeated MAT. Thrombolysis was used in these two patients. Complete symptomatic relief was achieved in 25 patients (96%) and partial relief in one. The hospital stay ranged from 2 to 4 days (mean 2.7 days). Recurrent thrombosis within the stent was observed in one case and recanalized with thrombolysis. The 1-year primary and secondary patency rate was 96% and 100%, respectively. No symptomatic pulmonary embolization, bleeding, and venous reflux were observed. Five patients complained about transitory low back pains during balloon angioplasty. CONCLUSION: Single-session endovascular treatment with MAT as the first-line thrombus removal method is feasible, safe, and effective for IVCS with secondary acute isolated iliofemoral DVT. Although limited, our experience suggests that patients thought to be at high risk of bleeding may be candidates for the present single-session endovascular protocol.

Effects of partial tonsillectomy on the immune functions of children with obstructive sleep apnea-hypopnea syndrome at early stage.

The purpose of this study was to investigate the changes in the humoral and cellular immunity of children with obstructive sleep apnea-hypopnea syndrome and hypertrophy of tonsils before and after plasma-mediated temperature-controlled radiofrequency ablation treatment. Fifty-seven children suffering from obstructive sleep apnea-hypopnea syndrome and with hypertrophy of tonsils were enrolled in this study. Thirty-seven children were grouped in the partial tonsillectomy group and 20, in the tonsillectomy group. The levels of CD3(+), CD4(+), CD8(+), and CD4(+)/CD8(+) were measured for cellular immunity, and the levels of IgG, IgA, and IgM were measured for humoral immunity. Blood samples were collected before and 1 and 3 months after the operation. The IgG, IgA, and IgM levels in the tonsillectomy group were significantly decreased 1 month after the operation, and recovered to the normal levels within 3 months of the operation (P < 0.05). However, the levels of IgG, IgA, and IgM in the partial tonsillectomy group decreased slightly, without a significant difference (P > 0.05). The cellular immunity of the 2 groups was not statistically different pre- and post-operation (P > 0.05). The results from the present study indicate that partial tonsillectomy by plasma-mediated temperature-controlled radiofrequency ablation did not impact on the humoral and cellular immunity of children.

Analysis of interaction between Bmhrp28 and BmPSI in sex-specific splicing of Bombyx mori Bmdsx gene.

Bombyx mori BmHRP28 and BmPSI, which belong to the family of RNA-binding proteins, have been identified binding to the female-specific exon 4 of the sex-determining gene Bmdsx pre-mRNA. However, the relationships between BmHRP28 and BmPSI still remain unclear. In this study, we carried out yeast two-hybrid (Y2H) and co-immunoprecipitation (Co-IP) analyses to address them. Y2H analysis showed that there was little or no direct binding between the BmHRP28 and BmPSI proteins. Also, the Co-IP experiments revealed that BmHRP28 and BmPSI coexisted in a multiprotein complex. Our results suggested that BmHRP28 and BmPSI form a muliprotein complex to regulate the splicing of Bmdsx pre-mRNA, but are not directly bound to each other. In an effort to find other regulatory factors in the multiprotein complex, we constructed a silkworm Y2H cDNA library of male early embryo. By Y2H screening, we identified an RNA-binding protein BmSPX, a putative component of the spliceosome, binding to BmPSI. These results indicated that BmHRP28 and BmPSI make up a spliceosome complex to regulate Bmdsx splicing and that BmSPX is another potential protein involved in this process. Our study provides some clues to better understand the mechanism of sex determination in the silkworm.

First Report of Citrus yellow vein clearing virus on Lemon in Yunnan, China.

Eureka lemon (Citrus limon) is an economically important crop in China. In 2009, a new disease was observed on Eureka lemon in an orchard of Ruili city in Yunnan Province. Nearly 10% of the plants were symptomatic with yellow vein clearing on lateral veins, combined with leaf crinkling and warping of young leaves. Water-soaked appearances were also seen on the adaxial leaf veins. The symptoms were present on mature leaves, resulting in almost 20% yield reduction. The symptoms on leaves were more severe in spring and autumn than in summer. This disease was transmitted by grafting from infected Eureka lemon to six lemon (C. limon) varieties (Femminello, Fino, Villafranca, Verna, Kutdiken, and Monachello) and two sour orange (C. aurantium) varieties (Daidai, Morocco), with inoculated plants showing similar symptoms. Variable symptoms were observed on other hosts, including severe yellowing and vein clearing on Dweet tangor (C. reticulata × C. sinensis), slight vein clearing on Tongshui 72-1 sweet orange (C. sinensis) and Marsh grapefruit (C. paradisi), but no symptoms were observed on Etrog citron (C. medica), Mexican lime (C. aurantifolia), or Ponkan (C. reticulata). When infected branches from Etrog citron, Mexican lime, and Ponkan were grafted onto healthy Morocco sour orange, typical yellowing and vein clearing symptoms were observed. In the mechanical transmission trials using eight herbaceous plant species, leaf rolling, shrinking, and pointed necrosis were observed on Vigna unguiculata, slight leaf rolling symptoms were observed on Capsicum frutescens, whereas Lycopersicum esculenum, Nicotiana benthamiana, Momordica charantia, Solanum melongena, Cucumis sativus, and Sesamum indicum remained symptomless. Symptom expression was influenced by temperature, being visible between 18 and 24°C but disappearing over 32°C. Crude extracts from young symptomatic lemon leaves were negatively stained for transmission electron microscopy observation (1) and showed filamentous virus particles of 13 to 15 × 400 to 1,000 nm. The described combination of symptoms in citrus species and herbaceous indicators, and virus morphology revealed by electron microscopy, suggested presence of Citrus yellow vein clearing virus (CYVCV). RNA extractions were made from 52 samples of Eureka lemon with yellow vein clearing symptoms. A one-step RT-PCR using primer pair (sense: 5'-TACCGCAGCTATCCATTTCC-3' and antisense: 5'-GCAGAAATCCCGAACCACTA-3'), designed from coat protein gene of CYVCV genome (GenBank Accession No. JX040635) (1), was performed. All of the symptomatic samples yielded the expected cDNA fragment and no amplicon was obtained from healthy plants. Three PCR amplicons were selected for cloning and sequencing. BLAST analysis showed that these nucleotide sequences (KJ859679 to 81) had greater than 97% nucleotide identity with the corresponding region of CYVCV reference genome (2), and therefore, this disease was associated with CYVCV. Symptoms resembling CYVCV infection were observed in China before (3); however, to our knowledge, this is the first proven report of CYVCV present in China. The identification of the pathogen now allows for appropriate field management and control measures. References: (1) V. Grimaldi et al. 13th IOCV Conference, 343-345, 1996. (2) G. Loconsole et al. Phytopathology 102:1168, 2012. (3) C. Zhou et al. 18th IOCV Conference, 44, 2010.

Coexistent gestational choriocarcinoma and mixed adenocarcinoma of the uterus.

OBJECTIVE: To report the first case of a uterine gestational choriocarcinoma coexisting with an endometrial carcinoma (EC) and to discuss its possible pathogenesis. MATERIALS AND METHODS: All tissues were examined histologically and monoclonal antibodies were used to evaluate the expression of HCG, HPL, P53, PTEN, and ER. Genotyping was performed on DNA extracted from the freshly dissected choriocarcinoma and the paraffin-embedded endometrial carcinoma along with parental blood DNA using multiplex STR-PCR at 16 loci. RESULTS: Histology identified two distinct tumors: a uterine tumor containing cytotrophoblastic and syncytiotrophoblastic cells and a second distinct neoplasm composed of adenocarcinoma resembling endometrioid and mucinous adenocarcinoma. Genotyping of the choriocarcinoma revealed alleles from both the patient and her husband and was classified as biparental in origin. The endometrial adenocarcinoma contained only maternal alleles and was thus classified as maternal in origin. CONCLUSIONS: This is the first description of the simultaneous diagnosis of a uterine gestational choriocarcinoma and an EC within the same patient. DNA genotyping and immunohistochemistry are valuable tools in distinguishing the different origins of coexisting tumors.