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HLA-DPB1*1553:01 differs from HLA-DPB1*09:01:01:01 by one nucleotide in exon 3., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-B*40:550 differs from HLA-B*40:01:02:01 by one nucleotide in exon 1., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Purpose: The objective of this study was to provide theoretically feasible strategies by understanding the relationship between the immune microenvironment and the diagnosis and prognosis of AML patients. To this end, we built a ceRNA network with lncRNAs as the core and analyzed the related lncRNAs in the immune microenvironment by bioinformatics analysis., Methods: AML transcriptome expression data and immune-related gene sets were obtained from TCGA and ImmPort. Utilizing Pearson correlation analysis, differentially expressed immune-related lncRNAs were identified. Then, the LASSO-Cox regression analysis was performed to generate a risk signature consisting immune-related lncRNAs. Accuracy of signature in predicting patient survival was evaluated using univariate and multivariate analysis. Next, GO and KEGG gene enrichment and ssGSEA were carried out for pathway enrichment analysis of 183 differentially expressed genes, followed by drug sensitivity and immune infiltration analysis with pRRophetic and CIBERSORT, respectively. Cytoscape was used to construct the ceRNA network for these lncRNAs., Results: 816 common lncRNAs were selected to acquire the components related to prognosis. The final risk signature established by multivariate Cox and stepwise regression analysis contained 12 lncRNAs engaged in tumor apoptotic and metastatic processes: LINC02595, HCP5, AC020934.2, AC008770.3, LINC01770, AC092718.4, AL589863.1, AC131097.4, AC012368.1, C1RL-AS1, STARD4-AS1, and AC243960.1. Based on this predictive model, high-risk patients exhibited lower overall survival rates than low-risk patients. Signature lncRNAs showed significant correlation with tumor-infiltrating immune cells. In addition, significant differences in PD-1/PD-L1 expression and bleomycin/paclitaxel sensitivity were observed between risk groups., Conclusion: LncRNAs related to immune microenvironment were prospective prognostic and therapeutic options for AML., Competing Interests: None., (© 2024 Published by Elsevier Ltd.)

HLA-B*15:659 differs from HLA-B*15:02:01:01 by one nucleotide in exon 2., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-DPA1*02:117 differs from HLA-DPA1*02:02:02:01 by one nucleotide in exon 2., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-DPB1*05:01:21 differs from HLA-DPB1*05:01:01:01 by one nucleotide in exon 3., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-B*40:555 differs from HLA-B*40:01:02:01 by one nucleotide in exon 3., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-C*03:651 differs from HLA-C*03:03:01:01 by one nucleotide in exon 4., (© 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

HLA-C*17:69 differs from HLA-C*17:01:01:02 by one nucleotide in exon 4., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Bryophytes represent a sister to the rest of land plants. Despite their evolutionary importance and relatively simple body plan, a comprehensive understanding of the cell types and transcriptional states that underpin the temporal development of bryophytes has not been achieved. Using time-resolved single-cell RNA sequencing, we define the cellular taxonomy of Marchantia polymorpha across asexual reproduction phases. We identify two maturation and aging trajectories of the main plant body of M. polymorpha at single-cell resolution: the gradual maturation of tissues and organs along the tip-to-base axis of the midvein and the progressive decline of meristem activities in the tip along the chronological axis. Specifically, we observe that the latter aging axis is temporally correlated with the formation of clonal propagules, suggesting an ancient strategy to optimize allocation of resources to producing offspring. Our work thus provides insights into the cellular heterogeneity that underpins the temporal development and aging of bryophytes., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Granulomatous lobular mastitis (GLM) is a rare and chronic benign inflammatory disease of the breast. Difficulties exist in the management of GLM for many front-line surgeons and medical specialists who care for patients with inflammatory disorders of the breast. This consensus is summarized to establish evidence-based recommendations for the management of GLM. Literature was reviewed using PubMed from January 1, 1971 to July 31, 2020. Sixty-six international experienced multidisciplinary experts from 11 countries or regions were invited to review the evidence. Levels of evidence were determined using the American College of Physicians grading system, and recommendations were discussed until consensus. Experts discussed and concluded 30 recommendations on historical definitions, etiology and predisposing factors, diagnosis criteria, treatment, clinical stages, relapse and recurrence of GLM. GLM was recommended as a widely accepted definition. In addition, this consensus introduced a new clinical stages and management algorithm for GLM to provide individual treatment strategies. In conclusion, diagnosis of GLM depends on a combination of history, clinical manifestations, imaging examinations, laboratory examinations and pathology. The approach to treatment of GLM should be applied according to the different clinical stage of GLM. This evidence-based consensus would be valuable to assist front-line surgeons and medical specialists in the optimal management of GLM., (© 2022. The Author(s).)

Objective: To explore the occurrence level of depressive symptoms and it's influencing factors among gas field workers. Methods: In October 2018, a cross-sectional study was conducted in 1726 gas field workers from a gas field by using cluster sampling method. Questionaire was used to evaluate the individual factors, depressive symptoms, occupational stress factors and stress regulatory factors. The correlation between depressive symptoms and occupational stress was analyzed. Multivariate logistic regression was used to analyze the influencing factors of depressive symptoms. Results: The depressive symptoms score of gas field workers was 12.00 (7.00, 19.00) point. Correlation analysis revealed that depressive symptoms score was positively related to sleep disorders ( r =0.598) , effort ( r =0.186) , daily tension ( r =0.478) , negative affectivity ( r =0.565) , social support ( r =0.446) and monotony of work (r=0.484) ( P <0.01) . And it was negatively related to reward ( r =-0.386) , work stability ( r =-0.294) , promotion opportunities ( r =-0.258) , positive affectivity ( r = -0.310) , self-efficacy ( r =-0.312) , contral strategy ( r =-0.268) , support strategy ( r =-0.209) and job satisfaction ( r =-0.398) ( P <0.01) . Multivariate logistic regression analysis revealed that sleep disorder, high negative affectivity, low support from colleagues, low support from family, high monotony of work and high daily tension were the risk factors for depressive symptoms of gas field worker ( OR =3.423, 95% CI : 2.644-4.397; OR =2.847, 95% CI : 2.200-3.683; OR =1.646, 95% CI : 1.215-2.116; OR =1.496, 95% CI : 1.164-1.923; OR =1.578, 95% CI : 1.227-2.303; OR =1.903, 95% CI : 1.480-2.440; P <0.01) . High work stability, high self-efficacy and high job satisfaction were protective factors for depressive symptoms of gas field workers ( OR =0.752, 95% CI : 0.591-0.958; OR =0.590, 95% CI : 0.465-0.749; OR =0.718, 95% CI : 0.516-0.999; P <0.05) . Conclusion: Occupational stress factors have a great influence on the depressive symptoms of gas field workers. Increased work stability, self-efficacy and job satisfaction could reduce the risk of depressive symptoms.

The behavior of γ-β/β 0 phase transition in TiAl alloy doped with β stabilizers (V, Cr, Mn) are studied by using the first principles method. It is found that alloying addition as well as anharmonic lattice vibration and disordered atomic occupation contributes to enhance the stability of cubic structure and accordingly introduce the disordered β phase into the high-temperature microstructure. The formation of low-temperature β 0 phase originates from not only the stabilization of cubic structure but also the destabilization of tetragonal structure. In particular, the latter is the main reason for the premature precipitation of the hard-brittle β 0 phase in the room-temperature microstructure at low nominal doping concentrations. We also find a special doping region in which the γ and the β phases are stable, while the β 0 phase is unstable. The existence of this region provides an opportunity for the regulation of the contents of β and β 0 phases.

Objective: To investigate the level of social support and its correlation with occupational stress among gas production workers in the field. Methods: In October 2018, the cluster sampling method was used to perform a cross-sectional survey for 1726 gas production workers in the field, and related data of these workers were collected, including age, education level, marital status, level of social support, and related factors for occupational stress. A Spearman's rank correlation analysis was used to investigate the correlation between social support and occupational stress, and the levels of occupational stress-related factors were compared between the groups with different social support scores. Results: The gas production workers in the field had a median (25th percentile, 75th percentile) social support score of 24.00 (19.00, 28.00) , and there was a significant difference in social support score between the workers with different posts or work shifts ( P <0.01) . Social support score was positively correlated with effort, daily stress, negative emotion, and job routinization ( P <0.05) and was negatively correlated with job satisfaction, reward, working stability, and promotion opportunity ( P <0.05) . The group with a high social support score had significantly higher scores of effort, job routinization, sleep disorders, and daily stress than the other two groups ( P <0.01) , and the group with a low social support score had significantly higher scores of reward, self-efficacy, positive affection, and job satisfaction than the other two groups ( P <0.01) . Conclusion: High-level social support plays an important role in alleviating occupational stress and protecting mental health among gas production workers in the field.

Objective: To explore the interaction between shift work and psychological capital on abnormal glucose and lipid metabolism. Methods: A convenient sampling survey of demographics characteristics, shift work and psychological capital was conducted on 1 415 natural gas field workers by questionnaire in October 2018,and their physiological and biochemical indexes were measured according to standard norms. Multivariate logistic regression model was used to analyze the interaction between shift work and psychological capital on abnormal glucose and lipid metabolism. Results: For 1 415 subjectsthe prevalence of abnormal blood glucose was 21.2%, the prevalence of diabetes was 8.3%.The prevalence of abnormal total cholesterol was 40.4%, the prevalence of hypercholesterolemia was 11.3%.The prevalence of abnormal triglyceride was 41.6%, the prevalence of hypertriglyceridemia was 24.7%.The detection rate of Low-density Lipoprotein was 17.3%, the detection rate of Low-density Lipoprotein was 4.0%, and the detection rate of high-density Lipoprotein was 1.3%. Multiple logistic regression model analysis showed that shift work, the low level of self-efficacy and the low level of optimism was positively associated with abnormal blood glucose, respectively ( P <0.05). Shift work was positively associated with abnormal triglyceride ( P <0.05). However, there was no interaction between shift work, low self-efficacy, low hope, low resilience, and low optimism on abnormal glucose and lipid metabolism. Conclusion: Shift work was a risk factor of abnormal blood glucose and triglyceride, self-efficacy and optimism were protective factors of abnormal blood glucose. There was no multiplicative interaction between shift work and psychological capital on abnormal glucose and lipid metabolism in the study population.

Objective: To explore the relationship between sleep quality and occupational stress in field gas recovery workers. Methods: In October 2018, cluster sampling method was adopted to conduct cross-sectional survey on 1726 field workers in a gas production oilfield. The individual characteristics, occupational stress factors, stress regulation factors, stress response and sleep quality, social support and coping strategies were evaluated by occupational stress measurement tools and job content questionnaire. Mann Whitney U test and Kruskal Wallis H test were used to compare sleep quality scores between the groups. Spearman rank correlation analysis was used to analyze the correlation between sleep quality and occupational stress, and logistic regression analysis was used to analyze multiple factors. Results: There were significant differences in sleep quality scores among different positions, gender, marital status, age, length of service, smoking and drinking ( P <0.05) . There were no significant differences in sleep quality scores between different education levels and work shift groups ( P >0.05) . Spearman rank correlation analysis showed that sleep quality score was negatively correlated with job satisfaction, reward, job stability, promotion opportunity, positive emotion, respect, self-esteem, control strategy, support strategy and self-efficacy score ( r(s) =-0.361, -0.311, -0.238, -0.261, -0.248, -0.212, -0.139, -0.188, -0.152, -0.226, P <0.01) , and was positively correlated with social support, giving, daily tension, negative emotion, work monotony and depression symptom ( r(s) =0.312, 0.279, 0.547, 0.493, 0.429, 0.599, P <0.01) . Compared with the high sleep quality score group, the middle and low sleep quality score groups had lower giving, work monotony, daily tension, depressive symptoms, negative emotions and social support ( P <0.01) , while the scores of respect, reward, job satisfaction, positive emotion, self-efficacy, job stability, promotion opportunity, control strategy and support strategy were higher ( P <0.01) . Multiple depressive symptoms, high daily tension, high negative emotion and high work monotony were the risk factors for sleep disorders ( OR =3.417, 2.659, 2.913, 1.543) . Conclusion: Depressive symptoms, daily tension and negative emotion have great influence on sleep quality of field gas recovery workers.

Objective: To investigate the effect of Toll-like receptor 7 (TLR7) in CD8(+) T cells activity from patients with breast cancer. Methods: Thirty-three patients with breast cancer, twenty-three patients with benign breast tumor, and twenty healthy individuals were collected from The First Affiliated Hospital of Xinxiang Medical University between December 2017 and March 2018. Peripheral blood mononuclear cells (PBMCs) were isolated, and CD8(+) T cells were purified. TLR7 protein and mRNA relative expression in CD8(+) T cells was measured using flow cytometry and real-time PCR, respectively. mRNA relative expressions corresponding to perforin, granzyme B, and FasL in CD8(+) T cells were measured in response to TLR7 agonist stimulation. Direct/indirect contact co-culture system of CD8(+) T cells and breast cancer cell line MCF-7 was also used to assess cytolytic and noncytolytic function in response to TLR7 agonist CL097 stimulation. Results: The mean fluorescence intensity corresponding to TLR7 protein in CD8(+) T cells from breast cancer patients was 124.0±15.32, which was significantly down-regulated in comparison with benign breast tumor patients (255.5±54.91) and healthy individuals (261.9±68.65) ( P< 0.000 1). TLR7 mRNA relative level was also remarkably reduced in CD8(+) T cells from breast cancer patients (1.97±1.18) in comparison with benign breast tumor patients (4.84±1.01) and healthy individuals (4.75±1.40) ( P< 0.000 1). TLR7 agonist CL097 stimulation notably increased mRNA relative levels of perforin and granzyme B mRNA in CD8(+) T cells ( P< 0.01), but not elevated FasL mRNA ( P> 0.05).Furthermore, TLR7 agonist CL097 stimulation enhanced the cytolytic and noncytolytic function of CD8(+) T cells to MCF-7 cells, which presented as the elevation of target cell death and increase of interferon-γ production in direct and indirect contact co-culture system. Conclusion: TLR7 agonist promoted CD8(+) T cells function from breast cancer patients.

Background: MicroRNAs are believed to influence breast cancer cell tumorgenicity by interacting with the production of tumor associated macrophages. At this stage, this hypothesis lacks sufficient empirical evidence. Our study is an investigation of the effects of let-7a on the function of human breast cancer cell lines that had undergone chemokine ligand 18 (CCL18) stimulation., Methods: Two breast cancer cell lines MDA-MB-231 and MCF-7 were transfected with let-7a mimics with or without CCL18 simulation. The expression level of let-7a was evaluated with qRT-PCR. Our study examined cell proliferation, migration and cell cycles following let-7a treatment. The predicted target of let-7a was identified and confirmed in vitro by a dual luciferase reporter system. The associations between let-7a, CCL18 and target gene expression were evaluated using RT-PCR and the Western blotting method., Results: The downregulated expression level of let-7a was observed in both breast cancer cell lines. When compared to the control and CCL18 stimulation groups, cell proliferation and migration in MDA-MB-231 and MCF-7 cells were significantly inhibited by let-7a. Furthermore, the cell cycle was dramatically blocked at the G2/M phase. The luciferase reporter identified Lin28 as the direct binding target of let-7a in both breast cancer cell lines., Conclusion: Upregulation of let-7a carries the potential to reverse CCL18 induced cell proliferation and migration alteration in breast cancer cells by regulating Lin28 expression. Our results provided evidence which suggests the use of let-7a as a therapeutic agent in the treatment of breast cancer., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

This study aims to observe the changes of myocardial injuries after the firearm wound-induced intestinal perforation in porcine abdomen. 42 healthy Landrace piglets were randomly divided into the control group and the injury group, which was then subdivided into the post-injury 1 h, 2 h, 4 h, 8 h, 12 h and 24 h subgroup. the LDH, CK and CK-MB levels of each group, as well as the plasma endotoxin, were determined and compared. The plasma endotoxin levels of the experimental groups were significantly higher than the control group, and the light microscope observation revealed that the 8 h, 12 h and 24 h subgroup appeared the gradually-aggravated myocardial cell edema and degeneration; the electron microscope revealed that the 4 h, 8 h, 12 h and 24 h subgroup appeared the mitochondrial swelling and dissolution gradually; the serum levels of LDH, CK and CK-MB of each experimental group were higher than the control group. The abdominal firearm wound-induced intestinal perforation would lead to the damaged changes of myocardial morphology and enzymes, which would aggravate as time went along.

Objective: To summarize the surgical effect and clinical application value of esophagectomy with extended 2-field lymph node dissection for patients with esophageal carcinoma., Methods: From June 1987 to December 2008, 1690 patients with esophageal cancer underwent esophagectomy with extended 2-field (thoracic and abdominal) dissection of lymph nodes. Patients with the middle and lower thoracic esophageal cancer underwent Ivor-Lewis esophagectomy, and patients with upper thoracic esophageal cancer underwent Akiyama esophagectomy. 2-field (thoracic and abdominal) lymph node metastases information and the 1, 3, 5, 10-year survival rates were analyzed retrospectively., Results: Lymph node metastases were found in 713 patients. The lymph node metastases rate was 42.2% (713/1690).Thoracic lymph node metastasis rate was 39.3% (665/1690), among which in the right pleural apical para-tracheal triangle was 20.7% (349/1690), in the posterior upper mediastinum was 26.3% (444/1690), in the lower mediastinum was 18.2% (307/1690). Abdominal lymph node metastasis rate was 20.1% (339/1690). THE Postoperative complication rate was 16.4% (278/1690), among which the pulmonary complication rate ranking the first, was 43.6% (136/312). The operative mortality rate was 0.2%. The 1-year, 3-year, 5-year and 10-year survival rates were 88.2% (1388/1574), 63.5% (868/1367), 54.8% (705/1287) and 30.8% (232/754), respectively. The 5-year survival rate in patients without lymph node metastasis was 76.2% (448/588), but that in patients with lymph node metastases was 36.8% (257/669)., Conclusion: The results of this study demonstrated that Ivor-Lewis and Akiyama esophagectomy with two-field lymph node dissection exposes the operation fields clearly and make radical lymphadenectomy thoroughly, especially the lymph nodes in the posterior upper mediastinum around the recurrent laryngeal nerve and in the right pleural apical para-tracheal triangle. It is essential that patients with esophageal carcinoma with lymph node metastases should undergo esophagectomy with extended 2-field dissection of lymph nodes. This can elevate the postoperative 5-year survival rate remarkably.

Background & Objective: As one of the principal causes of gene inactivation, aberrant hypermethylation in the promoter of cancer-related genes has attracted more and more attention. However, such studies on esophageal cancer are still limited. This study was to investigate the association between aberrant hypermethylation of MGMT gene and clinical characteristics as well as MTHFR C677T genetic polymorphisms in esophageal squamous cell carcinoma in a Chinese population., Methods: A molecular epidemiologic study was conducted at Yangzhong County, Jiangsu Province of China, on histologically confirmed esophageal squamous cell carcinoma patients who were operated in the People's Hospital of Yangzhong County between January 2005 and March 2006. Peripheral blood samples, esophageal cancer tissues and paracancerous normal tissues were collected. Methylation-specific polymerase chain reaction(MSP) was used to detect the CpG island methylation status of MGMT gene. Restrictive fragment length polymorphism (RFLP) technique was used to test polymorphisms of folate metabolism enzyme gene MTHFR. The association between methylation status of MGMT gene and clinical characteristics as well as MTHFR C677T polymorphisms were analyzed., Results: Among 125 esophageal squamous cell carcinoma patients, the aberrant hypermethylation rate of MGMT gene was 27.2% in cancer tissues and 11.2% in paracancerous normal tissues. No hypermethylation was found in normal esophageal tissues from 10 healthy adult subjects. Methylation rate of MGMT gene in cancer tissues was significantly higher in the patients with lymph node metastasis than in those without lymph node metastasis (37.3% vs. 18.2%, P=0.017). No association was found between aberrant DNA methylation and selected factors including sex, age, tobacco smoking, alcohol drinking and green tea drinking. After adjusting by potential confounders, variant allele of MTHFR C677T was found to be associated with hypermethylation of MGMT gene. Compared with wild type CC, the odds ratio was 3.34 (95% CI: 1.07-10.39) for CT and 3.83 (95% CI: 1.13-12.94) for TT., Conclusion: Aberrant CpG island hypermethylation of MGMT gene is closely related with the genesis and progression of esophageal squamous cell carcinoma.

Background & Objective: Regional lymph node metastasis plays an important role in the prognosis of esophageal carcinoma. However, the range of lymph node dissection is still controversial. This study was to investigate the regulations of lymph node metastasis of thoracic esophageal carcinoma in the mediastinum and upper abdomen, and explore the rational lymphadenectomy with Ivor-Lewis procedure., Methods: A total of 1 412 thoracic esophageal carcinoma patients underwent radical esophagectomy and mediastinal and abdominal lymphadenectomy by Ivor-Lewis procedure from 1990 to 2005 at Yangzhong People's Hospital; 517 of them underwent right para-trachea triangle field lymphadenectomy through the right pleural apical approach. The regulations of regional lymph node metastasis were analyzed., Results: Of the 1,412 patients, 323 (22.88%) had postoperative complications, 2 (0.14%) died during hospitalization, and 547 (38.74%) had lymph node metastasis. The lymph node metastasis rates were 32.30% in the right para-trachea triangle, 18.43% in the upper mediastinum, 5.31% in the lower mediastinum, and 17.28% in the upper abdomen(P<0.001). Of the 13 916 resected lymph nodes, 2 662 (19.13%) were positive; the metastasis degree (positive lymph nodes/resected lymph modes) were 23.83% in the right para-trachea triangle, 18.92% in the upper mediastinum, 21.07% in the lower mediastinum, and 17.20% in the upper abdomen. For those patients with the cancer focuses in the upper, middle and lower segments of the esophagus, the lymph node metastasis rates were 40.59%, 36.97% and 44.35% (P=0.093), respectively, while the lymph node metastasis degree in these 3 fields were 19.60%, 18.35%, and 21.82%, respectively. Both the lymph node metastasis rate and degree were significantly higher in the patients at advanced stage than in the patients at early stage (46.56% vs. 7.75%, 21.82% vs. 4.01%, P<0.001)., Conclusions: Regional lymph node metastasis, especially in the right para-trachea triangle and upper mediastinum, is a key factor for thoracic esophageal carcinoma. Ivor-Lewis esophagectomy with two-field lymph node dissection is a safe operation for thoracic esophageal carcinoma, and may increase the chances of complete resection.

The aim of this work was to identify the immunodominant regions of the Em18 antigen to improve the specificity in diagnosis of alveolar echinococcosis (AE). Two recombinant antigens ReEm18-1 and ReEm18-2, which have the same sequence except that nine amino acid residues are absent in ReEm18-2, were tested by ELISA and Western Blot (WB) for their diagnostic efficiency. Serological evaluation of the two antigens demonstrated that the sensitivity of both antigens was 95.5% in ELISA and WB, and the specificity was 93.6% and 95.7% in ELISA, and 81.4% and 82.9% in WB, respectively. Five more expression clones (EmS1-EmS5), which contain different regions of the Em18 sequence, were constructed for defining the immunodominant regions of the antigen. Fourteen monoclonal antibodies (mAbs) against ReEm18-2 antigen and the sera from different groups of patients were used to identify the epitope regions of the five antigen fragments. Results showed that the epitopes recognized by the mAbs are located in the N-terminal third of the sequence, but the immunodominant area recognized by native serum antibodies may be located further downstream (C-terminal) in the sequence. The nonspecific cross-reactivity is due to epitopes present in the C-terminal third of the sequence. The antigen fragments that contain the first two-thirds of the sequence have the same sensitivity to AE sera as those of the ReEm18-1 and ReEm18-2 antigens, but removal of the C-terminal third of the sequence improved the specificity of the assay from 93.6% to 99.3% (ELISA) and 81.4% to 90.7% (WB). We conclude that the necessary part of the ReEm18 antigen sequence for AE diagnosis is the N-terminal half to two-thirds of the entire sequence.

The human hookworm Necator americanus was maintained through one hundred generations in the golden hamsters. The strain is now routinely maintained in laboratory hamsters through serial passage, and is the laboratory strain of choice for vaccine studies. Comparison of the mitochondrial cytochrome oxidase 1 (cox-1) sequences was shown previously to be useful for comparing the genetic structure of populations of N. americanus in China. Cytochrome oxidase 1 genes were amplified by the polymerase chain reaction, and the sequences compared to those of N. americanus recovered from infected humans from several regions in China. Sequence comparison revealed little difference between the laboratory strain and the field isolates at the cox-1 locus, but also indicated that the laboratory strain is represented by a single cox-1 haplotype. These results suggest that the laboratory strain of N. americanus has undergone a severe genetic bottleneck, and that the genetic diversity in other genes, including potential vaccine antigens, could be similarly limited.

Objective: To clone, express and serologically evaluate the Em18 antigen gene of Echinococcus multilocularis for diagnostic purpose., Methods: Polymerase chain reaction (PCR) was employed for amplification of the target gene fragments which was then ligated with pET28a+ vector. The constructed plasmid was transferred into E. coli BL21 (DE3) for expression. The recombinant proteins were purified with Ni-NTA agarose by affinity chromatography. 237 sera were used for evaluating diagnostic value of the recombinant Em18 antigen., Results: Two high-level expression clones (designated as ReEm18-1 and ReEm18-2) were obtained. ReEm18-1 showed the expected sequence, ReEm18-2 showed the same sequence but with 27 nucleotides deletion. The molecular weight of the two expression proteins was Mr 28,000 and 26,000, respectively. Serological evaluation by ELISA was carried out using sera from 101 patients with alveolar echinococcosis (AE), 47 with cystic echinococcosis (CE), 30 with cysticercosis (CC), 10 with hepatic cancer (HC), 9 with schistosomiasis (Sj) and 40 from healthy persons (NH) from both endemic and non-endemic areas. The results showed an overall sensitivity of 86.1% and 90.1% with ReEm18-1 and ReEm18-2 for AE sera, specificity 93.4% and 94.1%, positive predictive value 90.6% and 91.9%, negative predictive value 90.1% and 92.8% and efficiency 90.3% and 92.4%, respectively. The correlation analysis between the size of AE lesions and the serum absorbance reacted with recombinant Em18 antigens showed that there was a positive correlation between antibody level and the course of the disease., Conclusion: ReEm18 antigens are specific for AE diagnosis, and the serum antibody level displays a good correlation with the course of the disease at early stage. Similar results achieved by both ReEm18-1 and ReEm18-2 antigens.

A cDNA encoding a surface-associated antigen was cloned from an Ancylostoma caninum infective larvae (L(3)) cDNA library by immunoscreening with pooled human immune sera. The sera were obtained from individuals living in an Ancylostoma duodenale hookworm-endemic region of China, who had light intensity infections and high antibody titers against A. caninum L(3). Ancylostoma caninum surface-associated antigen-1 is encoded by an 843 bp mRNA with a predicted open reading frame of 162 amino acids. Recombinant Ancylostoma caninum surface-associated antigen-1 was expressed in Escherichia coli and used to prepare a specific antiserum. A Western blot with anti-Ancylostoma caninum surface-associated antigen-1 specific antiserum showed that native Ancylostoma caninum surface-associated antigen-1 protein is expressed by both L(3) and adult hookworms; RT-PCR confirmed that the mRNA is transcribed in both stages. In adult hookworms, the protein localised to the basal layer of the cuticle and hypodermis of adult worms. Serological analysis determined that recombinant Ancylostoma caninum surface-associated antigen-1 protein is recognised by 61% of human sera from a Necator americanus hookworm endemic area in China, indicating the antigen is immunodominant. Anti-Ancylostoma caninum surface-associated antigen-1 antiserum partially inhibited (46.7%) invasion of hookworm L(3) into dog skin in vitro. Together these results suggest that Ancylostoma caninum surface-associated antigen-1 offers promise as a protective vaccine antigen.

Objective: To analyze antigens for searching specific antigenic components for immunodiagnosis of echinococcosis., Methods: Fourteen crude antigens from different tissues (cyst fluid, protoscoleces, laminated layer and germinal layer) of Echinococcus granulosus and E. multilocularis metacestodes and other 4 species of cestodes were analyzed by Western blotting. The differences of protein bands were compared for the 14 crude antigens by reacting with pooled sera from cystic echinococcosis (CE) and alveolar echinococcosis (AE) patients., Results: Eleven protein bands from the antigens reacted nonspecifically with sera from both CE and AE patients were Mr 130000, 100000, 94000, 80000, 75000, 66000, 62000, 52000, 38000, 32000, 24000. The highly specific protein bands recognized by AE sera were Mr 120000, 109000, 86000, 59000, 43000, 28000, 20000, 18000, and by CE sera were Mr 41000, 40000, 22000, 16000 and 12000., Conclusion: Different antigens shared by the two species of Echinococcus were examined and potential antigenic proteins specific for AE or CE sera were found, providing useful information for further identifying specific antigens for immunodiagnosis.

Objective: To investigate the incidence of lymph node metastasis (LNM) in the right para-tracheal triangle (RPT) of esophageal carcinoma patients and the technique of dissection., Methods: On the top of double mediastinal and abdominal lymphadenectomy, 333 esophageal carcinoma patients received RPT lymphadenectomy through the right pleural apical approach from 1990 to 2001., Results: In these 333 patients, the lymph node metastasis (LNM) rate in the RPT was 36.40%. A total of 457 nodes among 2 159 nodes removed gave a metastasis degree of 24.96%. The LNM rates in RPT for cervical, upper third, middle third, and lower third segments of esophagus were 66.67%, 45.45%, 34.19% and 15.79% (P < 0.05), while their respective metastasis degrees were 44.44%, 27.04%, 24.32% and 18.92% (P > 0.05). The frequency of positive nodes in the RPT for PTI, PT1, PT2, PT3 and PT4 was 0, 17.24%, 28.7%, 45.16% and 53.57%, while those of metastasis degree were 0, 8.77%, 17.62%, 33% and 41.17% (P < 0.01). The frequency of LNM in the RPT in papillary, erosive, patch-like and covert type of early tumor was 40%, 3.85%, 0 and 0 (P < 0.05), while those of the metastasis degree were 29.41%, 1.82%, 0 and 0 (P < 0.01). Higher rate of LNM in progressive stenotic esophageal carcinoma was observed compared with those of the other gross types (56.52%, P < 0.05), so was the degree (P < 0.01). The frequency of LNM in the RPT for mono-focal and multi-focal tumor was 34.98% and 70% without significant difference (P > 0.05), while the degree was 24.29% and 53.33% (P < 0.05). Postoperative complications were: leak (0.6%), and recurrent laryngeal nerve injury (1.2%). No injury of vein or infra-clavicular artery, tracheal damage or mortality occurred., Conclusion: 1. The lymph node metastasis from esophageal carcinoma has a tendency of wide spread and right para-tracheal triangle is an important region to be doomed. 2. With location, depth of tumor invasion and differentiation of tumor as major factors affecting LNM of esophageal carcinoma, dissection of this region should be paid more emphasis. 3. In early lesions, higher frequency of LNM in the RPT is found in papillary and erosive lesions than in the other macroscopic types. 4. Exposing the RPT, lymph node by dissection through a right pleural apical approach is very important and significant.

Objective: To search for the gene encoding specific antigen of Ancylostoma caninum that induces host's protective immunity., Methods: A lambda ZAP II cDNA library of A. caninum was screened with sera from dogs immunized subcutaneously with hookworm larvae(L3). After sequencing, insert gene (AcAg) from positive clones was ligated into PUC18 and PET28C. Recombinant pET28C plasmid was induced to expressed protein in the E. coli BL21. The characteristic of recombinant protein is analyzed by SDS-PAGE and Western blotting assay., Results: Five positive clones were obtained, and proved to be the same. The insert gene (AcAg) in pET28C vector expressed a recombinant protein of about 43 kDa. Using Western blotting assay, this protein was recognized by the sera from dog immunized with Ancylostoma caninum third-stage infected larvae and used for screening library., Conclusion: The AcAg, which exhibits 35% homologous to Caenorhabditis elegans gene unc-89, is a novel specific antigen of A. caninum. Its ability to elicit the protective immunity and the probability of the recombinant protein as a vaccine need to be further evaluated.

Objective: To detect and compare the COI gene sequences of Necator americanus collected from the provinces of Sichuan, Hainan, Yunnan, Hubei and Jiangsu, and to analyze the genetic diversity of the geographic isolates., Methods: COI genes of N. americanus were amplified from the genomic DNA by PCR and sequenced., Results: The COI gene sequences of N. americanus from five provinces were 97%-99% identical over 595 bp, and base variation occurred in 19 nucleotide sites in which the transition was more frequent than the transversion. The difference between the sequences ranged from 1.34% to 2.18%., Conclusion: The COI gene sequences show high identity among the geographic isolates of N. americanus with some difference at specific nucleotide sites.

Objective: To explore the factors of false negative antibody response in patients with echinococcosis granulosus (Eg) for improving immunodiagnosis., Methods: Indirect ELISA and sandwich ELISA were used to detect the specific antibody of IgG subclass (IgG1, IgG2, IgG3, IgG4) and IgA, IgM, IgE, as well as circulating antigen (CAg) and immunocomplex (CIC) in sera of Eg patients with negative response of total IgG., Results: Among 42 sera with IgG negative seroresponse, 32 were positive with IgG subclass, IgA, IgM or IgE antibody, 10 were negative in all 7 kinds of antibody response. The detection rate of specific IgG1, IgG4, IgA, IgM and IgE was 42.9%, 11.9%, 28.6%, 26.2% and 21.4% respectively, being significantly higher than in sera of the control. IgM level in children was higher than that in adults. IgG subclass in patients with liver Eg was higher than that of pulmonary hydatidosis, when testing with IgG1 combined one or two of the other six Ig antibodies. The highest positive rate (64.3%) was seen in IgG1 + IgA + IgM antibody system. The positive rate of CAg and CIC in IgG negative patients was 28.57% and 30.95%, respectively., Conclusion: The factors involved with seronegative response of total IgG in Eg patients might be low specific IgG, variant Ig antibody expression and formation of CIC. Combined detection of IgG1 + IgA + IgM could enhance the sensitivity of serological tests in Eg patients.

Objective: To detect serum antibody of cysticercosis by dot immunogold filtration assay for establishing a diagnostic kit for cysticercosis patients., Methods: Cyst fluid of cysticercus of Taenia solium after dialysis was used as diagnostic antigen in dot immunogold filtration assay and enzyme-linked immunosorbent assay to detect the serum antibody in patients with cysticercosis. Samples to be detected included 71 sera from patients with cysticercosis, 90 sera from healthy people, 20 sera from patients with other parasitic infections or brain tumor., Results: The sensitivity and specificity of dot immunogold filtration assay were 90.1%(64/71) and 95.6%(86/90), respectively. No positive reaction was recorded in cases with other diseases except one serum from a patient with brain tumor. The coincidence rate between dot immunogold filtration assay and enzyme-linked immunosorbent assay was 94.4% (152/161)., Conclusion: Dot immunogold filtration assay showed promising result for the diagnosis of cysticercosis.

Objective: To compare 2 kinds of ELISA methods for detecting circulating antigen in schistosomiasis patients., Methods: Monoclonal antibody-based dot-ELISA and sandwich ELISA., Results and Conclusion: The positive rates of dot-ELISA were higher than or similar to those of sandwich ELISA, the former being 48.0%-98.8% and the latter being 12.0%-93.8%. The specificities were 95.4%-100% and 90.0%, respectively.

The present paper deals with studies on the characteristics of Schistosoma japonicum isolated from five localities in the mainland of China. The following items were observed and compared including morphometric data, susceptibility of six mammalian hosts, prepatent period, compatibility between larvae and snail hosts, size of hepatic granuloma produced by eggs, immunoreactions in experimental animals, sensitivity to praziquantel, SDS-PAGE protein pattern and its antigenicity analysis, DNA hybridization and genetic variation and differentiation by analysis with multilocus enzyme electrophoresis. By means of these multidisciplinary methods, from morphological to molecular level, the following conclusions may be drawn from our results. The evidence indicates firstly that S. japonicum in the mainland of China comprises a strain complex with several components of geographically distributed strains. At least four distinct strains exist, ie Yunnan, Guangxi, Sichuan and Anhui-Hubei. Characteristics of each strain are distinct and the results of these studies lead to discussion on the problem of the intraspecific and interstrain differentiation of S. japonicum in the mainland of China.

Homogenates prepared from S. japonicum adult worms of different isolates from Anhui, Hubei, Guangxi, Yunnan and Sichuan Provinces were analyzed by SDS-PAGE and enzyme linked immunoelectrotransfer blot (EITB) tested with rabbit anti-snails antibody. The results of SDS-PAGE indicated that with silver staining both male and female worms of Guangxi isolate showed some definite differences in their protein profile, namely, absence of one band between 50-75 kDa in male worms and marked reduction in quantity of > 110 and 30 kDa bands in female worms. There was no obvious difference among other isolates both in male and female worms. The EITB patterns were similar in S. japonicum of Anhui and Hubei, and it was also the case with isolates from Yunnan and Sichuan, except that Yunnan female worms had a distinct band at 84 kDa which could hardly be seen in EITB pattern of Sichuan female worms; female worms of Guangxi isolates also showed a distinct 84 kDa band. The EITB pattern of male worms from Guangxi isolates showed 2 main bands of MW > 130 kDa against anti-Anhui snail antiserum which corresponded with the result of male worms of Anhui isolates. But these bands could not be seen with male worms from isolates of Yunnan and Sichuan.

A fragment of the ribosomal RNA gene of Schistosoma mansoni pSM889 and two DNA fragments specific to Echinococcus granulosus pHD5 and pEG18 have been used as DNA probes to assess the extent of genetic variability within E. granulosus. The DNA analysis, including restriction endonuclease digestion and Southern blot hybridization with the probes, did not demonstrate any genetic variation among E. granulosus collected from sheep in Xinjiang, Qinghai, Gansu and Ninxia. Similarly, there was no genetic variation among E. granulosus isolates collected from yak in Qinghai and Gansu provinces. The authors deemed that the yak isolates and the sheep isolates of E. granulosus appear to belong to a same strain.

Cloned DNA fragments pHD5, pSM889 and pEG18 have been used as DNA probes in the restriction endonuclease analysis and southern blot hybridization to characterize E. granulosus and E. multilocularis protoscolices from China. Southern blot hybridization method is sensitive, specific and has the advantage in identification over microscopic examination. The authors deem that it can be used in the base-line epidemiological survey and surveillance of hydatid disease to provide data for hydatid disease control.

The purpose of the present study was to investigate the SDS-PAGE separation of proteins of the 5 different isolates of Schistosoma japonicum from the mainland of China to determine their similarities and/or differences and to gain additional data which could give an added insight into the degree of relationship. Soluble proteins of freshly sonicated adult worm extract of the 5 different isolates (i.e., Anhui, Hubei, Guangxi, Sichuan and Yunnan) were compared by SDS-polyacrylamide gel electrophoresis. The results indicated that no marked difference could be observed among the isolates after gels were stained by Coomassie blue white both male and female worms of Guangxi isolate showed some definite differences in their protein profile, i.e., lack of 50-75 kDa band in male worms and a marked reduction in the quantity of > 110 and 30 kDa bands in female worms as shown by silver stain.

Antigens prepared from S. japonicum adult worms of different isolates i. e. Anhui, Hubei, Guangxi, Yunnan and Sichuan by origin were analyzed by enzyme linked immunoelectrotransfer blot (EITB) probed with rabbit anti-snail antibody (Figs 1,2). Anti-sera against Oncomelania h. hupensis from Anhui, Hubei, Yunnan and Sichuan localities were prepared separately and used in the tests. The EITB patterns were similar in S. j. isolates of Anhui and Hubei, and it was also the case among S. j. isolates of Yunnan and Sichuan except Yunnan female worms with a marked band of 84 kDa but it was almost invisible in EITB pattern of Sichuan female worms. Like Yunnan isolate, female worms of Guangxi isolate also showed marked 84 kDa bands. The EITB pattern of male worms from Guangxi isolate showed 2 main bands of mw > 130 kDa against anti-Anhui snail anti-serum, which corresponded with the male worms of Anhui isolate whereas the color of the bands was darker and denser in the former isolates, and these bands can not be seen in the male worms from isolates of Yunnan and Sichuan. Based on the above mentioned results in connection with the information about the susceptibility between different isolates of schistosomes and their snail hosts which we have reported before, some preliminary analysis and discussion were made.

Observations on the immunoreactivities toward egg antigens of different experimental animals infected with Schistosoma japonicum were made, which included mouse, rat, hamster, rabbit and rhesus monkey. The cercariae applied for infection were shed by snails collected from 5 different isolates, i.e., Anhui, Hubei, Sichuan, Yunnan and Guangxi in the mainland of China. Sera from the same sort of animals were separately pooled according to the different sources of infection mentioned above. Antibody reactivities were evaluated by COPT, LAT and ELISA. In COPT each serum sample was tested with ova from homologous and heterologous isolates, whereas soluble egg antigen merely prepared from Anhui isolate was used in LAT and ELISA. The level of antibodies detected by homologous or heterologous worm isolate antigens was compared. The results suggested that the positive antibody detection rate might not be influenced by antigens prepared from S. japonicum eggs of different isolates in the mainland of China.

The secondary cyst tissues derived from mice infected with protoscoleces of Echinococcus granulosus for 8-10 months were digested with 0.25% trypsin at 37 degrees C for 30 min. The separation of different cells in the remaining suspension was achieved by discontinuous gradient centrifugation. The germinal cells were washed 3 times with ice-cold HBSS, and then cultivated in the medium of RPMI 1640 supplemented with 20% of calf serum. The cells were kept in an incubator at 37 degrees C in an atmosphere of 95% air-5% CO2. After incubation for 5-7 days, the germinal cells began to multiply accompanied by the enlargement of cells as compared with those before incubation. The surface of both isolated and/or cultured cells showed smooth appearance examined by scanning electron microscopy. Immunofluorescence assay and enzyme-linked immunosorbent assay had been used for examining the specific antigenicity of the cells. The results showed that antigen components of E. granulosus were detected either on cell surface or in soluble proteins of the cells. Furthermore, 120 NIH female mice were inoculated intraperitoneally with 1-5 x 10(7) cultured germinal cells and sacrificed 1-3 months after inoculation. Only 2 cystic materials had been detected in two mice. Of which, one located in the liver and the other in peritoneal cavity of the animals. Histological examination noted that the cystic materials consisted of germinal layer and cyst fluid, but no laminated layer was observed. The above mentioned evidence demonstrated that the cells isolated from the cysts of E. granulosus were germinal cells.(ABSTRACT TRUNCATED AT 250 WORDS)

A monoclonal antibody (designated 8SE) against membrane antigen of adult Schistosoma japonicum labeled with HRP was used in dot-ELISA (direct method) to detect schistosomal circulating membrane antigen. Sera from 48 parasitologically confirmed schistosomiasis cases were tested, 39 (81.3%) were positive. No false positive reaction was found in sera from 24 healthy controls. No cross reaction was detected in sera from clonorchiasis or paragonimiasis in 18 cases each. This results suggest that circulating membrane antigen does exist in patients with schistosomiasis and it might be used as a complementary method for diagnosis of schistosomiasis. A preliminary result of idiotype/antiidiotype interaction inhibition reaction for detecting circulating membrane antigen was also presented.

Using EMBL3 phage DNA as a vector, a perfect genomic library of Echinococcus granulosus from sheep from Xinjiang has been constructed, which contains 1.2 x 10(6) independent recombinants. The main procedure of construction comprised: 1. extraction of Echinococcus granulosus genomic DNA, 2. partial digestion of the extracted E. granulosus genomic DNA with restriction enzyme Sau3Al, 3. harvest of 15-23 kb DNA fragments by electric elution equipment, 4. preparation of EMBL3 phage vector DNA, 5. cleavage of EMBL3 vector DNA with two restriction enzymes Bam HI and Eco RI to remove the central fragment, 6. ligation of the harvested 15-23 kb E. granulosus genomic DNA and the cleavaged vector DNA with T4 DNA ligase, 7. the package reaction of ligated recombinant DNA in vitro with the package protein and with Q359 strain (Spi-) to screen recombinants, 8. identification of the genomic library, 9. hybridization in situ by pSM889 probe, obtaining 6 positive recombinant clones. It was estimated that the E. granulosus genome was about 1.5 x 10(8) bp. A perfect E.g. genomic library is expected to contain 4.6 x 10(4) independent recombinants at minimum. The constructed genomic library has enough independent recombinants (1.2 x 10(6)). This is the first time to establish an E. granulosus genomic library in China. Previous work showed that there existed differences in many aspects, including pathogenicity, among various isolates of E. granulosus from different hosts and areas. We plan to employ this library to screen the E. granulosus intraspecies DNA probe by hybridization in situ. This kind of probe is envisaged to be of advantage for epidemiological investigation of the hydatid disease in China. It also provides a base for researching E. granulosus at the molecular level.

In the present paper, the results of detecting circulating antigen and/or antigen-antibody complexes by McAb against surface membrane antigen of adult Schistosoma japonicum were reported. The McAb, coded as 8SE4, was prepared by fusion of SP2/0 myeloma cells with spleen cells of the BALB/c mice immunized with the saline extract of adult S. japonicum. The 8SE4- directed antigen was proved to be located on the surface of the adult worm. After being purified by a DE52 column, 8SE4 was labelled with HRP and the conjugate (HRP-8SE4) was used in the test. For testing, the serum sample was first incubated with HRP-8SE4, then PEG (mw. 6,000) was added to precipitate the antigen-antibody complex. Upon centrifugation, OPD was added to the precipitate. Results were read by ELISA reader at 492nm. The OD value was found to be proportional to the amount of circulating antigen and/or antigen-antibody complexes. Results from 5 heavily infected (1,500-2,000 cercariae) rabbits showed that the OD values were raised significantly at the 6th week post infection, being 1.9-4.5 times higher than those before infection. The OD values of the 5 rabbits each lightly infected with 10-500 cercariae were also markedly raised 6 weeks post infection and reached the peak at the 8th week, then maintained in high levels until 11th week post infection. The worm burden of the 5 lightly infected rabbits were 4-326. No obvious correlations between OD values and worm loads were observed. The results suggested the existence of surface membrane-related antigen and/or antigen-antibody complexes in the circulation of infected rabbits.