[The 505th case: refractory ascites and monoclonal immunoglobulin].

A 52-year-old woman was admitted with a primary complaint of abdominal distension and increased abdominal circumference for more than half a year. There was no evidence of infection or solid tumor on abdominocentesis or laparoscopic surgery. Concurrently, smoldering multiple myeloma was diagnosed. Due to refractory ascites and portal hypertension, a transjugular intrahepatic portosystemic shunt was performed, but the efficacy was not satisfactory. As the anemia progressed, she was finally diagnosed with active multiple myeloma after monoclonal plasma cells were detected in the ascites by flow cytometry. Treated with a triplet regimen that included bortezomib, cyclophosphamide, and dexamethasone (BCD), she achieved a very good partial response and ascites regressed.

Comparison of patients with different cervical spine bone ages before and after arch expansion treatment based on cone-beam computed tomography.

OBJECTIVE: The changes of maxillary basal arch width, molar angle, palatal suture width and nasal cavity width were analyzed in patients with different cervical bone ages before and after maxillary rapid arch expansion treatment, providing more reference for orthodontic design and treatment in the future. PATIENTS AND METHODS: A total of 45 patients with maxillary lateral, insufficient development that underwent arch expansion treatment in Jiaxing Second Hospital between February 2021 and February 2022 were selected for the study. Patients were retrospectively grouped based on the cervical vertebra bone age, and divided into the pre-growth (15 cases), mid-growth (15 cases) and post-growth groups (15 cases). All patients had oral cone-beam computed tomography (CBCT) and lateral cranial radiographs taken before and after the treatment. Maxillary basal arch width, palatal suture width, nasal cavity width and molar angle were measured and analyzed using paired samples t-test, ANOVA and least significant difference test (LSD-T). RESULTS: The maxillary basal arch width, palatal suture width, nasal cavity width and molar angle in the three groups were significantly changed after arch expansion treatment (p<0.05). There was no statistically significant difference in all measurement indexes between patients in the pre-growth and the mid-growth groups (p>0.05), but there was statistically significant difference between patients in the pre-growth and the late-growth groups (p<0.05). There were statistically significant differences in all measurement indexes between the middle-growth and the late-growth group (p<0.05). CONCLUSIONS: Rapid expansion of arch can be used to enlarge the width of palatal suture, maxillary basal arch, and nasal cavity in adolescent patients of different bone ages. With the increase of cervical bone age, the bony effect of expansion of arch gradually decreases, while the dental effect increases. Appropriate overcorrection should be made during arch expansion in late growth and excessive tooth tilt should be avoided to conceal bony width irregularities.

[Short-term efficacy and safety of vedolizumab in patients with inflammatory bowel disease].

Objective: To evaluate the short-term efficacy and safety of vedolizumab in patients with inflammatory bowel disease (IBD). Methods: Patients with moderate and severe active IBD at the first use of vedolizumab from May 1 to October 31, 2021 were retrospectively enrolled. Then the clinical characteristics, and the efficacy and safety of vedolizumab were evaluated. Meanwhile, the clinical response rate, biological response rate and endoscopic response rate were calculated. Multivariate analysis was used to evaluate the independent influencing factors of short-term clinical efficacy and safety. Results: A total of 78 patients (44 males and 34 females) with IBD were enrolled, with a mean age of (40.5±11.9) years. The clinical remission rate, clinical response rate, biological remission rate, biological response rate and endoscopic remission rate was 60.3% (47/78), 85.9% (67/78), 70.5% (55/78), 43.6% (34/78) and 47.0% (31/66) respectively after 14 weeks of treatment. Body mass index (BMI) ≥ 18.5 kg/m2 (HR=5.04, 95%CI: 1.50-16.91, P=0.009) and biological remission at 6 weeks of treatment (HR=15.22, 95%CI: 3.16-73.38, P=0.001) were predictors of endoscopic remission at 14 weeks of treatment. Adverse reactions occurred in 57 patients, with an incidence of 73.1%. The main manifestations were liver and kidney damage (37.2%) and infection (26.9%). Conclusions: More than half of patients with moderate and severe active IBD can achieve clinical remission after 14 weeks of vedolizumab treatment. Baseline BMI level and biological remission at 6 weeks of treatment are predictors of mucosal healing at 14 weeks. The incidence of adverse reactions is not low, although serious adverse reactions are rare in short-term treatment.

[The 491st case: thrombocytosis, gastrointestinal hemorrhage, intestinal perforation].

Essential thrombocythemia (ET) is a chronic myeloproliferative neoplasm (MPN) featured by clonal proliferation of platelets, thrombosis and hemorrhage. Portal hypertension is a serious complication of ET associated with poor prognosis. We report a patient with ET complicated with acute upper gastrointestinal hemorrhage and intestinal perforation due to portal hypertension. She had an uneventful recovery after surgical and endoscopic treatment.

Comparison of rapid maxillary expansion and pre-fabricated myofunctional appliance for the management of mouth breathers with Class II malocclusion.

OBJECTIVE: Pre-fabricated myofunctional appliances and rapid maxillary expansion (RME) has been used for the treatment of mouth-breathers with Class-II malocclusion. This study aimed to compare the treatment effects of hyrax and pre-fabricated myofunctional appliance (T4K) for the management of mouth breathers with Class II Malocclusion in mixed dentition stage. PATIENTS AND METHODS: Case records of mouth breathers with Class II Division 1 malocclusion patients treated at our institute with T4K or hyrax appliance between June 2015 to May 2019 were retrieved. The Pancherz analysis was used to compare the treatment effects. RESULTS: Data of 28 patients (14 in each group) were compared. Significant advancement of maxilla was seen in both groups while mandibular length improved only with the T4K appliance. SNA and SNB changes were significantly greater in the T4K group. Molar relationship improved in both groups. Molar correction was obtained by 55.6% skeletal change and 44.4% dental change with RME. In the T4K group the corresponding values were 48.1% and 51.9% respectively. CONCLUSIONS: Our results suggest that both pre-fabricated myofunctional appliance and RME are suitable for the treatment of mouth breathers with Class II malocclusion in the mixed dentition period. Sagittal correction of maxilla and mandible may be somewhat better with the T4K appliance. Although the dental compensation may be slightly more with the T4K appliance and it may inhibit the skeletal remodeling.

What do patients really want? An in-depth examination of patient experience in four Australian hospitals.

BACKGROUND: Patient satisfaction is an important outcome measure guiding quality improvement in the healthcare setting while the patient-centred care movement places increasing importance on patient engagement in clinical decision-making. However, the concept of patient satisfaction is not clearly defined, and beliefs of patients are not always evident in health surveys. Researchers rarely follow up on surveys to explore patient views and what they mean in greater depth. This study set out to examine perceptions of hospital care, through in-depth, qualitative data capture and as a result, to gather rich, patient-driven information on user experience and satisfaction in the Australian healthcare setting; and identify influencing factors. METHODS: Focus groups were undertaken in four St Vincent's Health Australia (SVHA) hospitals in 2017 where participants discussed responses to eight questions from the Press Ganey Patient Experience Survey. Thirty people who were inpatients at SVHA. RESULTS: Good communication and high-quality information at arrival and discharge were found to be important to patients. Communication breakdown was also evident, further exacerbated by a range of environmental factors such as sharing a room with others. Overall, patients' felt that while their spiritual needs were well-supported by the hospital staff at all SVHA hospitals, it was the clinical teams prioritised their emotional needs. Good communication and environments can improve patient experience and follow-up at home is vital. CONCLUSIONS: Patient-centred care needs careful planning with patients involved at entry and exit from hospital. Focused communication, environmental changes, attending to complaints, and clearer discharge strategies are recommended.

Evaluation of Clinical Relevance of Drug-Drug Interaction Alerts Prior to Implementation.

INTRODUCTION: Drug-drug interaction (DDI) alerts are often implemented in the hospital computerized provider order entry (CPOE) systems with limited evaluation. This increases the risk of prescribers experiencing too many irrelevant alerts, resulting in alert fatigue. In this study, we aimed to evaluate clinical relevance of alerts prior to implementation in CPOE using two common approaches: compendia and expert panel review. METHODS: After generating a list of hypothetical DDI alerts, that is, alerts that would have been triggered if DDI alerts were operational in the CPOE, we calculated the agreement between multiple drug interaction compendia with regards to the severity of these alerts. A subset of DDI alerts (n = 13), with associated patient information, were presented to an expert panel to reach a consensus on whether each alert should be included in the CPOE. RESULTS: There was poor agreement between compendia in their classifications of DDI severity (Krippendorff's α: 0.03; 95% confidence interval: -0.07 to 0.14). Only 10% of DDI alerts were classed as severe by all compendia. On the other hand, the panel reached consensus on 12 of the 13 alerts that were presented to them regarding whether they should be included in the CPOE. CONCLUSION: Using an expert panel and allowing them to discuss their views openly likely resulted in high agreement on what alerts should be included in a CPOE system. Presenting alerts in the context of patient cases allowed panelists to identify the conditions under which alerts were clinically relevant. The poor agreement between compendia suggests that this methodology may not be ideal for the evaluation of DDI alerts. Performing preimplementation review of DDI alerts before they are enabled provides an opportunity to minimize the risk of alert fatigue before prescribers are exposed to false-positive alerts.

[The effect of WT1 expression on the prognosis of allogeneic hematopoietic stem cell transplantation in acute leukemia].

Objective: To study the effect of WT1 expression on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acute leukemia (AL) and its significance as molecular marker to dynamically monitor minimal residual disease (MRD) . Methods: Retrospectively analyzed those AL patients who underwent allo-HSCT in the First Hospital Affiliated to Zhejiang University School of Medicine during Jan 2016 to Dec 2017, a total number of 314 cases, 163 males and 151 females, median age was 30 (9-64) years old. Comparing the difference of WT1 expression at diagnosed, pre-HSCT and after HSCT. Using the receiver operating characteristic (ROC) curve to determine the WT1 threshold at different time so as to predict relapse. The threshold of WT1 expression before transplantation was 1.010%, within 3 months after HSCT was 0.079% and 6 months after HSCT was 0.375%. According to these thresholds, WT1 positive patients were divided into low expression groups and high expression groups. Analyzed the relationship between overall survival (OS) , disease-free survival (DFS) , cumulative incidence of relapse (CIR) and WT1 expression. Results: The OS and DFS of high expression group pre-HSCT were lower than low expression group [69.2% (9/13) vs 89.1% (57/64) , χ(2)=4.086, P=0.043; 53.8% (7/13) vs 87.5% (56/64) , χ(2)=9.766, P=0.002], CIR was higher than low expression group [30.8% (4/13) vs 7.8% (5/64) , P=0.017]. There was no significant difference of OS and DFS between high expression and low expression group of 3 months after HSCT (P=0.558, P=0.269) . The OS and DFS of high expression group of 6 months after transplantation were both lower than low expression group (P=0.049, P=0.035) . Multivariate analysis showed that WT1>0.375% when 6 months after transplantation was the only independent prognostic factor for shorter DFS (P=0.022) . There was no statistically significant difference in CIR between the high-expression group and the low-expression group 3 months after transplantation and 6 months after transplantation (P=0.114, P=0.306) . Conclusion: High expression of WT1 before and after HSCT was an adverse prognosis factor. It is of clinical practical value to use WT1 as a transplant recommendation index for patients with acute leukemia and as a marker to monitor MRD dynamically.

LncRNA CCAT1 as the unfavorable prognostic biomarker for cholangiocarcinoma.

OBJECTIVE: To investigate the expression of long noncoding RNA CCAT1 in cholangiocarcinoma (CCA) and to assess the CCAT1 expression as a prognostic biomarker for CCA. PATIENTS AND METHODS: The CCAT1 expression in tumor tissues and paired adjacent normal tissues from 91 CCA patients was detected by quantitative real-time PCR. The association of the CCAT1 expression with clinicopathological features of CCA patients and the prognostic value of the CCAT1 expression for overall survival was also evaluated by Kaplan-Meier, Cox regression model and ROC analysis. RESULTS: The CCAT1 expression was significantly upregulated in CCA tumor tissues compared with adjacent normal tissues. The CCAT1 expression was obviously associated with histological differentiation, lymph node invasion, and TNM stage. The overall survival of CCA patients with high CCAT1 expression was worse. Furthermore, the CCAT1 expression could be considered as an independent prognostic factor in predicting the overall survival for CCA patients. CONCLUSIONS: Our study showed that lncRNA CCAT1, which was upregulated and associated with aggressive malignant behavior, may serve as a novel prognostic biomarker and potential therapeutic target for cholangiocarcinoma.

Detecting the potential cancer association or metastasis by multi-omics data analysis.

Comprehensive multi-omics data analyses have become an important means for understanding cancer incidence and progression largely driven by the availability of high-throughput sequencing technologies for genomes, proteomes, and transcriptomes. However, how tumor cells from the site of origin of the cancer begin to grow in other sites of the body is very poorly understood. In order to examine potential connections between different cancers and to gain an insight into the metastatic process, we conducted a multi-omics data analysis using data deposited in The Cancer Genome Atlas database. By combining somatic mutation data along with DNA methylation level and gene expression level data, we applied a Bayesian network analysis to detect the potential association among four distinct cancer types namely, Head and neck squamous cell carcinoma (Hnsc), Lung adenocarcinoma (Luad), Lung squamous cell carcinoma (Lusc), and Skin cutaneous melanoma (Skcm). Further validation based on the 'identification of somatic signatures' and the 'association rules analysis' confirmed these associations. Previous investigations have suggested that common risk factors and molecular abnormalities in cell-cycle regulation and signal transduction predominate among these cancers. This evidence indicates that our study provides a rational analysis and hopefully will help shed light on the links between different cancers and metastasis as a whole.

[Detecting shared pathways linked to rheumatoid arthritis with other autoimmune diseases in a in silico analysis].

Pathway-based analysis approach has exploded in use during the last several years. It is successful in recognizing additional biological insight of disease and finding groupings of risk genes that represent disease developing processes. Therefore, shared pathways, with pleiotropic effects, are important for understanding similar pathogenesis and indicating the common genetic origin of certain diseases. Here, we present a pathway analysis to reveal the potential disease associations between RA and three potential RA-related autoimmune diseases: psoriasis, diabetes mellitus, type 1 (T1D) and systemic lupus erythematosus (SLE). First, a comprehensive knowledge mining of public databases is performed to discover risk genes associated with RA, T1D, SLE and psoriasis; then by enrichment test of these genes, disease-related risk pathways are detected to recognize the pathways common for RA and three other diseases. Finally, the underlying disease associations are evaluated with the association rules mining method. In total, we identify multiple RA risk pathways with significant pleiotropic effects, the most unsurprising of which are the immunology related pathways. Meanwhile for the first time we highlight the involvement of the viral myocarditis pathway related to cardiovascular disease (CVD) in autoimmune diseases such as RA, psoriasis, T1D and SLE. Further Association rule mining results validate the strong association between RA and T1D and RA and SLE. It is clear that pleiotropy is a common property of pathways associated with disease traits. We provide novel pathway associations among RA and three autoimmune diseases. These results ascertain that there are shared genetic risk profiles that predispose individuals to autoimmune diseases.

Unit cohesion, traumatic exposure and mental health of military personnel.

BACKGROUND: The benefit of military unit cohesion to morale and psychological resilience is well established. But it remains unclear whether unit cohesion modifies the association between deployment-related traumatic exposure and mental health problems. AIMS: To examine the association between unit cohesion, traumatic exposure and poor mental health [symptoms of post-traumatic stress disorder (PTSD), psychological distress and alcohol dependency] and assess whether the relationship between traumatic exposure and poor mental health differs by level of unit cohesion. METHODS: A self-reported cross-sectional survey of Australian military personnel deployed to Iraq or Afghanistan between 2001 and 2009. RESULTS: Among 11411 participants, those with low levels of unit cohesion had higher odds of PTSD symptoms [aOR (95% CI): 2.54 (1.88, 3.42)], very high psychological distress [aOR (95% CI): 4.28 (3.04, 6.02)] and a high level of alcohol problems [aOR (95% CI): 1.71 (1.32, 2.22)] compared with those reporting high unit cohesion on deployment. Higher exposure to traumatic events on deployment was associated with greater risk of PTSD symptoms, very high levels of psychological distress and high levels of alcohol problems in this cohort. However, there was no evidence of a statistically significant interaction between unit cohesion and traumatic exposures in influencing poor mental health. CONCLUSIONS: Our findings suggest that both unit cohesion and traumatic exposure are independently associated with poor mental health. Efforts to improve military unit cohesion may help to improve the mental health resilience of military personnel, regardless of their level of traumatic exposure.

Factors associated with low unit cohesion in Australian Defence Force members who deployed to the Middle East (2001-2009).

INTRODUCTION: Unit cohesion has been shown to bolster the mental health of military personnel; hence, it is important to identify the characteristics that are associated with low unit cohesion, so that interventions to improve unit cohesion can be targeted and implemented. Little is known about the factors associated with low unit cohesion. This research aims to identify demographic, military service and deployment factors associated with low unit cohesion. METHODS: Data from a self-reported cross-sectional study of 11 411 current or ex-serving Australian military personnel deployed to Iraq or Afghanistan between 2001 and 2009 were used. Multivariable logistic regression was used to investigate the research aims. RESULTS: Being female (adjusted OR (aOR) (95% CI) 1.35 (1.21 to 1.51)), non-commissioned officer (aOR (95% CI) 1.50 (1.39 to 1.62)), lower ranked (aOR (95% CI) 1.74 (1.51 to 2.01)) or having left military service (aOR (95% CI) 1.71 (1.46 to 2.02)) was associated with reporting low unit cohesion. Potentially modifiable factors such as performing logistic roles on deployment (aOR (95% CI) 1.13 (1.01 to 1.27)), dissatisfaction with work experience on deployment such as working with colleagues who did not do what was expected of them (aOR (95% CI) 4.09 (3.61 to 4.64)), and major problems at home while deployed (aOR (95% CI) 1.50 (1.38 to 1.63)) were also associated with reporting low unit cohesion. CONCLUSIONS: This is the first study to identify demographic, military service and deployment factors associated with low unit cohesion. The modifiable nature of unit cohesion means that military leaders could use this information to identify subgroups for targeted resilience interventions that may reduce vulnerabilities to mental health problems and improve the job satisfaction, preparedness and deployment experiences of serving members.

Immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody.

We examined the immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody. Genomic DNA from the M5 strain of goat Brucella was amplified by polymerase chain reaction and cloned into the prokaryotic expression vector pGEX-4T-1. The expression and immunological characteristics of the fusion protein GST-omp31 were subjected to preliminary western blot detection with goat Brucella rabbit immune serum. The Brucella immunized BALB/c mouse serum was detected using purified protein. The high-potency mouse splenocytes and myeloma Sp2/0 cells were fused. Positive clones were screened by enzyme-linked immunosorbent assay to establish a hybridoma cell line. Mice were inoculated intraperitoneally with hybridoma cells to prepare ascites. The mAb was purified using the n-caprylic acid-ammonium sulfate method. The characteristics of mAb were examined using western blotting and enzyme-linked immunosorbent assay. A 680-base pair band was observed after polymerase chain reaction. Enzyme digestion identification and sequencing showed that the pGEX-4T-1-omp31 prokaryotic expression vector was successfully established; a target band of approximately 57 kDa with an apparent molecular weight consistent with the size of the target fusion protein. At 25°C, the expression of soluble expression increased significantly; the fusion protein GST-omp31 was detected by western blotting. Anti-omp31 protein mAb was obtained from 2 strains of Brucella. The antibody showed strong specificity and sensitivity and did not cross-react with Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, or Bacillus pyocyaneus. The pGEX-4T-1-omp31 prokaryotic expression vector was successfully established and showed good immunogenicity. The antibody also showed strong specificity and good sensitivity.

Donor TLR9 gene tagSNPs influence susceptibility to aGVHD and CMV reactivation in the allo-HSCT setting without polymorphisms in the TLR4 and NOD2 genes.

Owing to ethnicity of the population, those best confirmed polymorphisms in the TLR (toll-like receptor)4 and NOD2 genes with significantly prognostic impact on allogeneic hematopoietic SCT (allo-HSCT) seem to be more applicable to Europeans and are nonpolymorphic in the Asian population. The influence of innate immunity gene polymorphisms on the outcomes of allo-HSCT in those populations has been questioned. We evaluated the influence of 10 candidate single nucleotide polymorphisms (SNPs) in the TLR1, TLR2, TLR3, TLR8 and TLR9 genes on the outcomes of allo-HSCT in a Chinese population including 138 pairs of patients and unrelated donors and a second cohort of 102 pairs of patients and HLA-identical sibling donors. We found that two tagSNPs in the TLR9 gene in the donor side, +1174 A/G (rs352139) and +1635 C/T (rs352140), influenced the risk of acute GVHD (aGVHD) and CMV reactivation. Furthermore, the presence of the susceptible haplotype (A-C) in donor may be an informative predicator of worse OS at 5 years compared with those with the G-C and G-T haplotypes (58% vs 82.9%, P=0.024). Our data suggested an unrecognized association between donor TLR9 tagSNPs and the risk of HSCT-related complications in a population without polymorphisms in the TLR4 and NOD2 genes.

Levels of effectiveness of gene therapies targeting survivin and its splice variants in human breast cancer cells.

In order to develop an effective strategy of breast cancer therapy targeting survivin and its splice variants survivin-ΔEx3 and survivin-2B, the present study constructed four expression vectors by fusing the survivin antisense gene, the survivin (T34A) gene, the survivin-ΔEx3 antisense gene, and the survivin-2B gene with the enhanced green fluorescent protein (eGFP) gene. Each of these vectors was transiently transfected into the B-Cap-37 human breast cancer cell line. The effects of these four vectors with diverse genes on the proliferation and apoptosis of B-Cap-37 breast cancer cells were examined and compared in vitro using MTT and flow cytometry assays. Results of the MTT assay indicated that all four gene therapy plasmids were most effective at inhibiting the proliferation of B-Cap-37 cells 72 h after transfection. However, the four gene therapies had different rates of cell inhibition. pcDNA3.1(+)-egfp-anti-survivin and pcDNA3.1(+)-survivin (T34A)-egfp had almost equivalent or better effectiveness at suppressing cell growth. pcDNA3.1(+)-egfp-anti-survivin-ΔEx3 moderately inhibited the growth of B-Cap-37 cells. In contrast, pcDNA3.1(+)-survivin-2B-egfp had limited inhibition of cell growth. Similar profile of effectiveness of four gene therapies in soliciting cell apoptosis was also observed. These results suggest the relative importance of targeting survivin and its splice variant survivin-ΔEx3 in breast cancer treatment.

The beneficial impact of missing KIR ligands and absence of donor KIR2DS3 gene on outcome following unrelated hematopoietic SCT for myeloid leukemia in the Chinese population.

The effect of natural killer (NK) cell alloreactivity on the outcome of unrelated hematopoietic SCT (HSCT) remains a topic of debate. NK cell alloreactivity after allogeneic HSCT is regulated by killer-cell Ig-like receptors (KIRs). To investigate the influence of KIRs on outcome after unrelated HSCT, we retrospectively analyzed the HLA and KIR genotypes of 116 donor-recipient pairs. We found that missing KIR ligands in recipients were significantly associated with a decreased leukemic relapse risk (P=0.019, HR=0.329), mainly in myeloid disease (P=0.003, HR=0.193). This beneficial effect was seen in AML/myelodysplastic syndrome and also in chronic myeloid leukemia. In myeloid disease, missing KIR ligands also improved 5-year OS (P=0.034, HR=0.430) and disease-free survival (DFS) (P=0.024, HR=0.445). Meanwhile, the presence of donor-activating KIR2DS3 gene was associated with increased relapse risk (P=0.003, HR=5.046), decreased OS (P=0.004, HR=3.181) and DFS (P=0.003, HR=2.919) in myeloid disease. No effect was seen in patients with lymphoid disease. Our study indicated that, in unrelated HSCT for myeloid leukemia, missing KIR ligands in recipients offered a lower relapse risk and a long-term survival advantage. The presence of KIR2DS3 in the donor was an important risk factor for myeloid leukemia.

Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells.

Epidermolysis bullosa acquisita is an autoimmune blistering disease characterized by circulating and skin basement membrane-bound IgG autoantibodies to type VII collagen, a major structural protein of the dermal-epidermal junction. Regulatory T cells (T(reg)) suppress self antigen-mediated autoimmune responses. To investigate the role of T(reg) in the the autoimmune response to type VII collagen in a mouse model, a monoclonal antibody against mouse CD25 was used to deplete T(reg). A recombinant mouse type VII collagen NC1 domain protein and mouse albumin were used as antigens. SKH1 mice were used as a testing host. Group 1 mice received NC1 immunization and were functionally depleted of T(reg); group 2 mice received NC1 immunization and rat isotype control; and group 3 mice received albumin immunization and were functionally depleted of T(reg). Results demonstrated that anti-NC1 IgG autoantibodies with high titres, as determined by enzyme-linked immunosorbent assay and Western blotting, developed in all mice immunized with NC1 (groups 1 and 2), but were undetected in group 3 mice. The predominant subclasses of anti-NC1 autoantibodies were IgG1, IgG2a and IgG2b; furthermore, these antibodies carried only the kappa light chain. IgG autoantibodies in the sera of NC1-immunized mice reacted with mouse skin basement membrane in vitro and deposited in skin basement membrane in vivo as detected by indirect and direct immunofluorescence microscopy, respectively. Our data suggest that the development of autoimmunity against type VII collagen in mice is independent of T(reg) function and the autoimmune response is mediated by both Th1 and Th2 cells. We speculate that the basement membrane deposition of IgG may eventually lead to blister development.