[Clinicopathological features of esophageal carcinoma with ductal differentiation of esophageal gland].

Objective: To investigate the clinicopathological characteristics of esophageal carcinoma with gland duct differentiation. Methods: The clinical, morphologic and immunohistochemical (IHC) features of eight cases of esophageal carcinoma with gland duct differentiation diagnosed from 2012 to 2022 at the First Affiliated Hospital of Soochow University were summarized. Results: There were four males and four females, with a mean age of 68.5 (range 59-82) years. Two tumors were located in middle esophagus, five in the lower esophagus, and one in the cardia. The mean diameter was 2.4 cm (range 0.6-4.5 cm). The tumor had a bilayer epithelial structure, including the inner luminal epithelium and the outer basal epithelium. Immunohistochemistry showed that CK7 (8/8) and CK18 (8/8) were positive in the inner epithelium. p40 (8/8), p63 (8/8) and CK5/6 (8/8) were positive in the outer epithelium. SMA, calponin and CD117 were all negative. p53 mutants were found in all eight cases (strong and diffuse positivity in 6/8; complete loss of expression in 2/8). No columnar metaplasia, intestinal metaplasia and ectopic gastric mucosa were observed in the surface squamous epithelium in the cases. The mean follow-up time was 21.5 months (range 5-51 months). Seven patients survived and one patient died 31 months after surgery due to recurrence and liver metastasis. Conclusion: Esophageal carcinoma with esophageal gland duct differentiation is a rare tumor with unique histologic and IHC characteristics.

[Methylated SDC2 testing in stool DNA for early screening of colorectal cancer in Shipai Town, Dongguan City].

Objective: To explore the utility of stool-based DNA test of methylated SDC2 (mSDC2) for colorectal cancer (CRC) screening in residents of Shipai Town, Dongguan City. Methods: This was a cross-sectional study. Using a cluster sampling method, residents of 18 villages in Shipai Town, Dongguan City were screened for CRC from May 2021 to February 2022. In this study, mSDC2 testing was employed as a preliminary screening method. Colonoscopy examination was recommended for individuals identified as high-risk based on the positive mSDC2 tests. The final screening results, including the rate of positive mSDC2 tests, the rate of colonoscopy compliance, the rate of lesions detection, and the cost-effectiveness of screening, were analyzed to explore the benefits of this screening strategy. Results: A total of 10 708 residents were enrolled and completed mSDC2 testing, giving a participation rate of 54.99% (10 708/19 474) and a pass rate of 97.87% (10 708/10 941). These individuals included 4 713 men (44.01%) and 5 995 women (55.99%) with a mean age of (54.52±9.64) years. The participants were allocated to four age groups (40-49, 50-59, 60-69, and 70-74 years), comprising 35.21%(3770/10 708), 36.25% (3882/10 708), 18.84% (2017/10 708), and 9.70% (1039/10 708) of all participants, respectively. mSDC2 testing was positive in 821/10 708 (7.67%) participants, 521 of whom underwent colonoscopy, resulting in a compliance rate of 63.46% (521/821). After eliminating of 8 individuals without pathology results, data from 513 individuals were finally analyzed. Colonoscopy detection rate differed significantly between age groups (χ2=23.155, P<0.001),ranging from a low of 60.74% in the 40-49 year age group to a high of 86.11% in the 70-74 year age group. Colonoscopies resulted in the diagnosis of 25 (4.87%) CRCs, 192 (37.43%) advanced adenomas, 67 (13.06%) early adenomas, 15 (2.92%) serrated polyps, and 86 (16.76%) non- adenomatous polyps. The 25 CRCs were Stage 0 in 14 (56.0%) individuals, stage I in 4 (16.0%), and Stage II in 7(28.0%). Thus, 18 of the detected CRCs were at an early stage. The early detection rate of CRCs and advanced adenomas was 96.77% (210/217). The rate of mSDC2 testing for all intestinal lesions was 75.05% (385/513). In particular, the financial benefit of this screening was 32.64 million yuan, and the benefit-cost ratio was 6.0. Conclusion: Screening for CRCs using stool-based mSDC2 testing combined with colonoscopy has a high lesion detection rate and a high cost-effectiveness ratio. This is a CRC screening strategy that deserves to be promoted in China.

[Development of Clinical practice guidelines: procedure and standard].

In the last decade, the development of clinical practice guidelines in China has grown rapidly. However, with regards to the guidelines that have been established in the past, few were of high quality and in line with international standards. The main reason for this was that many clinical experts were not familiar with the procedures and rules of clinical guidelines before established, which lowered the quality seriously. Clinical practice guidelines are based on a clinical problem that is distilled into populations, interventions, comparison and outcome (PICO). After comprehensive systematic review, recommendations are made through evidence grading and strength of recommendation system. In addition, other issues should be noted such as pros and cons of the recommendation for specific population, preferences and values of the population, cost-effectiveness, and the health care system. A high-quality guideline requires multidimensional thinking (from clinicians, patients and policy makers), the implementation of a standard procedure (to ensure guidelines scientifically sound, honest and transparent), as well as the collaboration of multiple organizations (including experts, methodologists and policy makers).

[Early efficacy of islet transplantation in the treatment of adult advanced diabetes].

Objective: To evaluate the safety and efficacy of islet transplantation for patients with advanced diabetes. Methods: Five cases of islet allotransplantation were performed on 4 adult recipients. The same blood type adult brain-dead pancreas donors were selected and the islets were prepared in GMP laboratory. The prepared islet suspension was slowly injected into the liver of the recipients within 30-60 minutes. The immunosuppressive regimen was a combination of basiliximab, tacrolimus and mycophenolate mofetil and TNF-alpha monoclonal antibody was used to reduce the post-transplant inflammatory response. Insulin was temporarily applied to control blood glucose after surgery, and the dosage of insulin was adjusted to decrease according to the blood glucose level until it was discontinued. Results: A total of 5 islet transplants were performed in 4 patients, including 1 patient who received the second islet transplantations. All operations were succeed and the blood glucose and portal pressure were stable during the operation. Exogenous insulin was continued to keep blood glucose level stable (4-12 mmol/L) after surgery. Four cases (including the one who received two islet transplantation) started to stop using insulin at the third to fourth week, and the insulin dosage of the other case was 74% lower than that before the operation, and no hypoglycemic reaction occurred in all patients after islet transplantation. The C-peptide level in 3 patients reached the normal range, and the level in one patient with type I diabetes (without insulin release) remained at 0.45-0.6 µg/L (0.15-0.2 nmol/L). In addition, one patient showed a rise in blood glucose again and continued to use insulin half a year after insulin discontinuation. Then, he was performed the second islet transplantation which showed good effect and stopped taking insulin in 10 days after surgery. There were 3 cases of liver puncture bleeding after opeation, of which 2 cases were treated with ultrasound radiofrequency ablation to stop bleeding, 1 case stopped spontaneously, and no other complications were found. Conclusions: Islet transplantation is effective in the treatment of advanced diabetes patients with small trauma and high safety, which is worthy of more promotion. Long-term efficacy and maintenance therapy still need further investigation.

[Effect of Wnt/ß-catenin signal regulating EMT level on the differentiation of mouse ESC to liver tissue].

Objective: To study embryonic stem cell (ESC) differentiation into liver tissue structure from the perspective of epithelial mesenchymal transition (EMT). Methods: ESC of Balb/c mice was selected to induced into hepatic cell using hepatocyte growth factor (HGF), fibroblast growth factor (FGF) in vitro, and at the time points of metaphase (13 d) and maturity (17 d) of differentiation, dynamic inhibition of Wnt/ß-catenin signal was made to reduce the level of EMT. Finally, three-dimensional organization structure growth of the differentiation cells was observed in the differentiation system.Expressions of the liver cells vascular markers[albumin (Alb) and vascular endothelial growth factor (VEGFR)]were detected. Results: During the differentiation of ESC, the level of early EMT in the experimental group and the control group was not significantly different. The level of mid-late EMT in the experiment group was significantly lower than the control group. On the day 18 and 20 of differentiation, the relative mRNA expression level of E-cadherin was 0.61±0.15 and 0.47±0.05 in the experimental group, and 0.07±0.05 and 0 in the control group, respectively.The expression level of ALB/AFP/CK8/CK19 in the experimental group was generally higher than that of the control group in the same period, while CD31/VEGFR1 markers in the experimental group decreased more slowly in the late period of differentiation compared with the control group. In the supernatant of ESC culture, the Alb of the experimental group could be detected onday 7, and the concentration was (0.32±0.02) mg/L, while Alb in the control group was (0.19±0.05) mg/L. Urea in the experimental group could be detected on the day 13, and the concentration was (8.7 ±1.0) µmol/L, and the urea concentration of the control group was (3.1±1.2) µmol/L. The concentration of Alb and urea in the culture supernatant of ESC differentiation system increased significantly with the prolongation of the differentiation time, and the Alb and urea concentrations in the experimental group were significantly higher than those in the control group at the same time period. In addition, the differentiated cells in the experimental group could maintain the growth of three-dimensional tissue, while the differentiated cells in the control group eventually showed a single cell state. The expression of hepatic and vascular cell markers could be detected in the experimental group. Immunofluorescence results showed that the hepatocytes and vascular structures were tightly arranged. HE staining showed the formation of hepatic lobular structure, while the control group had no vascular component markers. Conclusion: The differentiation of ESC into liver tissue can be effectively promoted by decreasing the level of EMT at the mid-late stage of ESC differentiation.

[The efficacy comparation of adenoidectomy with acupuncture and tympanonstomy in children secretory otitis media].

Objective:This study aims to the comparative study of AT+A (adenoidectomy with acupuncture) and AT+T (adenoidectomy with tympanonstomy tube) to monitor and compare the therapeutic effect and prognosis of secretory otitis media in children. The study make a summary and give the clinical suggestions as well.Method:We collected and analyzed 280 outpatients of children secretory otitis media from March 2015 to March 2016.Among them,172 cases took the adenoidectomy with acupuncture and 108 cases took the adenoidectomy with tympanonstomy tube. This research used the therapeutic effect indicators,middle ear effusion time and one year follow-up to evaluate the pros and cons of two surgery methods in different areas.Result:The patients of both groups had relatively good therapeutic effect which promoted with time. There were no significant difference between AT+A and AT+T in tympanic membrane. While AT+T group acted better than AT+A group in pure tone average and tympanum figure. The middle ear effusion time of AT+T group was significantly shorter than AT+A group. In one year follow-up, there were no difference in hearing loss between two groups.But AT+T group performed better in recurrence rate, infection rate and total rate.Conclusion:Since the adenoidectomy with tympanonstomy tube method has a lot of advantages over adenoidectomy with acupuncture,it's better to use AT+T in severechildren secretory otitis media when situation is available.

Development and Validation of a Nomogram for Predicting Incidence of Early Allograft Dysfunction Following Liver Transplantation.

OBJECTIVE: Early allograft dysfunction (EAD) is frequent complication post-liver transplantation and is closely related to recipient's mortality and morbidity. We sought to develop a nomogram for predicting incidence of EAD. METHODS: Based on multivariate analysis of donor, recipient, and operation data of 199 liver transplants from deceased donors between 2013 and 2015, we identified 5 significant risk factors for EAD to build a nomogram. The model was subjected to prospective validation with a cohort of 42 patients who was recruited between January and June 2016. The predictive accuracy and discriminative ability were measured by area under the receiver operating characteristic curve (AUC). The agreement between nomogram prediction and actual observation was showed by the calibration curve. RESULTS: Incidence rate of EAD in the training set and validation cohort were 55.91% (104/199) and 54.76% (23/42), respectively. In the training set, according to the results of univariable and multivariable analysis, 5 independent risk factors including donor gender, donor serum gamma-glutamyl transpeptidase level, donor serum urea level, donor comorbidities (respiratory, cardiac, and renal dysfunction), and recipient Model for End-stage Liver Disease score were identified and assembled into the nomogram. The AUC of internal validation using bootstrap resampling and prospective validation using the external cohort of 42 patients was 0.74 and 0.60, respectively. The calibration curves for probability of EAD showed acceptable agreement between nomogram prediction and actual observation. According to the score table, the probability of EAD was under 30% when the total point tally was under 72. But when the total was up to 139, the risk of EAD increased to 60%. CONCLUSION: We've established and validated a nomogram that can provide individual prediction of EAD for liver transplant recipients. The practical prognostic model may help clinicians to qualify the liver graft accurately, making a more reasonable allocation of organs.

[The effect of semimature dendritic cell and the levels of Treg on transplantation tolerance of hepatocytes differentiated from mouse embryonic stem cell].

Objective: To investigate the inducing effect and mechanism of semimature dendritic cell (smDCs) on transplantation tolerance of hepatocytes differentiated from mouse embryonic stem cells (ESCs), and to study the connections between smDCs and regulatory dendritic cells (regDCs). Methods: ESCs of 129 mouse labelled green fluorescent protein (GFP) were induced to hepatocytes by using previous methods. Meanwhile, bone marrow mononuclear cells of 129 mouse were induced to smDCs and regDCs. Moreover, the hepatocytes differentiated from 129 mouse ESCs were transplanted into liver of BALB/c mouse 3 days after infusing smDCs and regDCs suspension of 129 mouse into BALB/c mouse by tail vein respectively. After that, the growth status and survival time of transplanted cells in the recipient and infiltration of lymphocytes in transplant sites were observed. Furthermore, Foxp3 expression of peripheral blood CD4+ T cells was also tested. Results: In the control group, the transplanted cells in liver of BALB/c mouse survived only about 1 week. In contrast, the transplanted cells of smDC groups and regDCs groups survived about 4 weeks and the transplant sites of smDC groups also had less CD3(+) T cells. The morphology of smDCs were similar with regDCs. The expression of MHC-Ⅱ, CD40, CD80 and CD86 on smDCs and regDCs were moderate. Moreover, the Foxp3 expression of peripheral blood CD4+ T cells in smDC groups was higher than that in the control groups, from 1.11% up to 5.38%. The Foxp3 expression in regDC groups rose to 3.87%. Conclusion: The smDCs could induce transplantation tolerance of hepatocytes differentiated from 129 mouse ESCs in the recipient. The mechanism was associated with high level of Foxp3(+) Tregs, which could be increased by means of smDCs appropriate expression of MHC-Ⅱ, CD40, CD80 and CD86. The smDCs and regDCs were the same type of tolerance dendritic cells.

[The effects and expression of Survivin and Aurora-b in laryngeal carcinoma].

Objective:The aim of this study is to investigate the relationship between the expression of Survivin and Aurora-b in laryngeal carcinoma and its clinical and pathological features, and to determine the possibility of Survivin and Aurora-b being a biomarker in laryngeal carcinoma diagnosis or being an predictor of overall survival (OS) in laryngeal carcinoma patients underwent surgical resection. Method:Eighty-six cases of laryngeal carcinoma tissues diagnosed by pathology were collected, and 86 cases of adjacent tissues and 22 cases of normal tissues were selected as control. The expression of Survivin and Aurora-b were detected by immunohistochemistry in laryngeal carcinoma, adjacent tissues and normal tissue. Result:Survivin and Aurora-b in laryngeal carcinoma were significantly higher than those in adjacent tissues and normal tissues (P<0.01). The expression of Survivin protein was correlated with clinicopathological features such as lymph node metastasis and recurrence (P<0.05), but it was not related to the pathological features such as age, sex, clinical stage and differentiation. The expression of Aurora-b was correlated with the pathological grade clinical stag (TNM stage) and recurrence in laryngeal carcinoma (P<0.05). Survival curves show Survivin and Aurora-b in laryngeal cancer is associated with prognosis of laryngeal carcinoma. The result of COX regression analysis show that the expression of Aurora-b and recurrence are independent prognostic risk factors for postoperative patients with laryngeal carcinoma (P<0.05), while Survivin and lymph node metastasis are not independent prognostic factors influencing prognosis. Conclusion:Our study shows that Survivin and Aurora-b and their interaction may play an important role in the metastasis and prognosis of laryngeal carcinoma. The expression levels of Survivin and Aurora-b might be biomarkers for laryngeal carcinoma diagnosis and an independent predictor of OS after surgical resection, which would provide a novel insight into diagnosis and therapy of laryngeal carcinoma.

[Association between 23 urinary metals and mean platelet volume among a community-dwelling population in Wuhan, China].

OBJECTIVE: To investigate the potential association between 23 urinary metals and mean platelet volume (MPV) among a community population in Wuhan. METHODS: A total of 3 053 community residents who lived in the sampling buildings for more than 5 years, aged from 18 to 80 years, were recruited using a stratified, cluster sampling approach in Wuhan city, China. Blood and urine samples were obtained from participants in the morning under fasting conditions. Urinary metals, including aluminum, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, arsenic, selenium, rubidium, strontium, molybdenum, cadmium, tin, antimony, barium, tungsten, thallium, lead and uranium, were measured by inductively coupled plasma mass spectrometry. The MPV contents were determined using a fully automated clinical chemistry analyzer. Participants with missing data on covariates or cardiovascular disease were excluded. According to the reference intorvals of MPV for Chinese adults, the participants were classified into normal (7.0-11.0 fl) and high MPV (>11.0 fl) subgroups. Data from 2 203 participants were used to evaluate the associations between urinary metals and MPV levels using generalized linear regression models, and the risk of abnormal elevation of MPV using multivariable logistic regression models. The false discovery rate (FDR)-corrected P-value from 23 hypothesis tests was used to adjust for multiple testing. RESULTS: After adjusting for potential confounders, urinary concentrations of arsenic (P50=2.431 µg/mmol creatinine) and molybdenum (P50=4.035 µg/mmol creatinine) were significantly associated with increased MPV levels and the risk of abnormal elevation of MPV. In contrast, urinary aluminum (P50=2.706 µg/mmol creatinine) and thallium (P50=0.046 µg/mmol creatinine) were associated with decreased MPV levels, but also the risk of abnormal elevation of MPV. The regression coefficients and 95% CIs were 0.119 (0.043-0.196) for arsenic (FDR-adjusted P=0.018), 0.119 (0.042-0.195) for molybdenum (FDR-adjusted P=0.018), -0.115 (-0.195--0.034) for aluminum (FDR-adjusted P=0.029), and -0.307 (-0.386- -0.228) for thallium (FDR-adjusted P<0.001), respectively. When comparing the extreme quartiles for arsenic, molybdenum, aluminum and thallium, adjusted OR and the 95%CIs were 1.830 (1.382-2.423), 1.496 (1.125-1.989), 0.566 (0.412-0.779) and 0.302 (0.219-0.416), respectively, and FDR-adjusted P-values were <0.001,<0.014,<0.008 and<0.001, respectively. Moreover, significant associations were found between an increased risk of abnormal MPV elevation with urinary iron (P50=6.716 µg/mmol creatinine) , antimony (P50=0.014 µg/mmol creatinine) and uranium (P50=0.003 µg/mmol creatinine) , and a decreased risk with urinary tungsten (P50=0.010 µg/mmol creatinine) and lead (P50=0.265 µg/mmol creatinine) . When comparing the extreme quartiles for iron, antimony and uranium, the respective adjusted OR (95%CI) were 1.866 (1.395-2.496), 1.507 (1.111-2.043) and 1.452 (1.063-1.984), and the respective FDR-adjusted P-values were <0.001,<0.022 and<0.012. The respective adjusted OR (95%CI) were 0.551 (0.417-0.726) and 0.534 (0.394-0.725), and the respective FDR-adjusted P-values were<0.001 and<0.001, when comparing the extreme quartiles for tungsten and lead. Based on multi-metal models, urinary chromium (P50=0.120 µg/mmol creatinine) and selenium (P50=0.646 µg/mmol creatinine) were associated with increased risk of abnormal MPV, while urinary nickel (P50=0.193 µg/mmol creatinine) was associated with decreased risk of abnormal MPV. When comparing the extreme quartiles for chromium, selenium and nickel, adjusted OR (95% CI) were 1.578 (1.054-2.363), 1.718 (1.159-2.549) and 0.535 (0.373-0.767), respectively, and the FDR-adjusted P-values were 0.017, 0.028 and 0.002, respectively. CONCLUSION: In the general population of Wuhan city, exposure to aluminum, chromium, iron, nickel, arsenic, selenium, molybdenum, antimony, tungsten, thallium, lead and uranium were all associated with abnormal MPV elevation.

[Strategies in clinical diagnosis and treatment of steroid-resistant acute rejection after orthotopic liver transplantation].

OBJECTIVE: To explore the diagnostic evidence and treatment strategies for steroid-resistant acute rejection (SRAR) after orthotopic liver transplantation. METHODS: A retrospective analysis was performed among 1038 patients undergoing orthotopic liver transplantation in our hospital from January 2004 to December 2013. A total of 169 acute rejection (AR) episodes occurred in 153 patients. Sixteen of the patients were diagnosed with SRAR because of no response to large-dose steroid pulse therapy. The diagnosis and treatment of the 16 patients were analyzed retrospectively. Comparison of data was made by χ2 test or t test, and a P value of <0.05 was considered to be significant. RESULTS: The incidence of AR after liver transplantation was 14.74% (153/1038) in all the patients. The incidence of SRAR was 9.47% (16/169) in patients with AR. In the 16 patients with SRAR, 3 were treated with anti-CD3 monoclonal antibody (OKT3), 9 were treated with monoclonal antibody against IL-2 receptor, and 4 received antithymocyte globulin (ATG) therapy. After treatment, SRAR was reversed in 12 of the 16 patients and caused death of the other 4 patients, yielding a reversal rate of 75% and a mortality rate of 25%. CONCLUSION: SRAR after liver transplantation has a low incidence rate but poor prognosis. The diagnosis of SRAR is mainly based on the clinical manifestation, laboratory test, liver biopsy, and poor response or rejection to methyl prednisolone pulse therapy. ATG and OKT3 achieve substantial outcomes in most of the patients in the treatment of SRAR. Particularly, compared with OKT3, ATG achieves a higher reversal rate and fewer adverse reactions, which is expected to become the first-line treatment of SRAR.

Endogenous interleukin-22 protects against inflammatory bowel disease but not autoimmune cholangitis in dominant negative form of transforming growth factor beta receptor type II mice.

During chronic inflammation, interleukin (IL)-22 expression is up-regulated in both CD4 and CD8 T cells, exerting a protective role in infections. However, in autoimmunity, IL-22 appears to have either a protective or a pathogenic role in a variety of murine models of autoimmunity and, by extrapolation, in humans. It is not clear whether IL-22 itself mediates inflammation or is a by-product of inflammation. We have taken advantage of the dominant negative form of transforming growth factor beta receptor type II (dnTGF-ßRII) mice that develop both inflammatory bowel disease and autoimmune cholangitis and studied the role and the biological function of IL-22 by generating IL-22(-/-) dnTGF-ßRII mice. Our data suggest that the influence of IL-22 on autoimmunity is determined in part by the local microenvironment. In particular, IL-22 deficiency exacerbates tissue injury in inflammatory bowel disease, but has no influence on either the hepatocytes or cholangiocytes in the same model. These data take on particular significance in the previously defined effects of IL-17A, IL-12p40 and IL-23p19 deficiency and emphasize that, in colitis, there is a dominant role of IL-23/T helper type 17 (Th17) signalling. Furthermore, the levels of IL-22 are IL-23-dependent. The use of cytokine therapy in patients with autoimmune disease has significant potential, but must take into account the overlapping and often promiscuous effects that can theoretically exacerbate inflammation.

Serum microRNAs as novel biomarkers for primary sclerosing cholangitis and cholangiocarcinoma.

The diagnosis of primary sclerosing cholangitis (PSC) is difficult due to the lack of sensitive and specific biomarkers, as is the early diagnosis of cholangiocarcinoma (CC), a complication of PSC. The aim of this study was to identify specific serum miRNAs as diagnostic biomarkers for PSC and CC. The levels of 667 miRNAs were evaluated in 90 human serum samples (30 PSC, 30 CC and 30 control subjects) to identify disease-associated candidate miRNAs (discovery phase). The deregulated miRNAs were validated in an independent cohort of 140 samples [40 PSC, 40 CC, 20 primary biliary cirrhosis (PBC) and 40 controls]. Receiver operating characteristic (ROC) curves were established and only miRNAs with an area under the curve (AUC) > 0·70 were considered useful as biomarkers. In the discovery phase we identified the following: 21 miRNAs expressed differentially in PSC, 33 in CC and 26 in both in comparison to control subjects as well as 24 miRNAs expressed differentially between PSC and CC. After the validation phase, miR-200c was found to be expressed differentially in PSC versus controls, whereas miR-483-5p and miR-194 showed deregulated expression in CC compared with controls. We also demonstrate a difference in the expression of miR-222 and miR-483-5p in CC versus PSC. Combination of these specific miRNAs further improved the specificity and accuracy of diagnosis. This study provides a basis for the use of miRNAs as biomarkers for the diagnosis of PSC and CC.

High-dimensional immune profiling of total and rotavirus VP6-specific intestinal and circulating B cells by mass cytometry.

In-depth phenotyping of human intestinal antibody secreting cells (ASCs) and their precursors is important for developing improved mucosal vaccines. We used single-cell mass cytometry to simultaneously analyze 34 differentiation and trafficking markers on intestinal and circulating B cells. In addition, we labeled rotavirus (RV) double-layered particles with a metal isotope and characterized B cells specific to the RV VP6 major structural protein. We describe the heterogeneity of the intestinal B-cell compartment, dominated by ASCs with some phenotypic and transcriptional characteristics of long-lived plasma cells. Using principal component analysis, we visualized the phenotypic relationships between major B-cell subsets in the intestine and blood, and revealed that IgM(+) memory B cells (MBCs) and naive B cells were phenotypically related as were CD27(-) MBCs and switched MBCs. ASCs in the intestine and blood were highly clonally related, but associated with distinct trajectories of phenotypic development. VP6-specific B cells were present among diverse B-cell subsets in immune donors, including naive B cells, with phenotypes representative of the overall B-cell pool. These data provide a high dimensional view of intestinal B cells and the determinants regulating humoral memory to a ubiquitous, mucosal pathogen at steady-state.

Analysis of ectoparasites (chigger mites, gamasid mites, fleas and sucking lice) of the Yunnan red-backed vole (Eothenomys miletus) sampled throughout its range in southwest China.

The Yunnan red-backed vole Eothenomys miletus (Rodentia: Cricetidae) is an endemic rodent species and reservoir host of zoonoses in southwest China. Based on a large host sample (2463 voles collected from 39 localities between 2001 and 2013), a general analysis of four categories of ectoparasite (fleas, sucking lice, chigger mites and gamasid mites) on E. miletus across its entire range of distribution was made. This analysis identified a total of 71 895 ectoparasites belonging to 320 species (30 species of flea, 9 of sucking louse, 106 of gamasid mite and 175 of chigger mite) with a high prevalence (87%), mean abundance (29.19) and mean intensity (33.69). Of the 18 vector species of zoonoses found on E. miletus, the flea Ctenophthalmus quadratus (Siphonaptera: Hystrichopsyllidae) and chigger mite Leptotrombidium scutellare (Trombidiformes: Trombiculidae) were the dominant species; these are the main vectors of zoonoses in China. All of the dominant parasite species showed an aggregated distribution pattern. Male voles harboured more species of parasite than females. Chigger mites represented the most abundant species group on voles and their prevalence was positively correlated with mean abundance (r = 0.73; P < 0.05). As a single rodent species, E. miletus has a high potential to harbour abundant ectoparasites with high species diversity and high rates of infestation. The sex of the vole affects ectoparasite infestation.

Kidney transplantation from cardiac death donors with terminal acute renal failure.

OBJECTIVES: Kidney transplantation from donation after cardiac death (DCD) donors with terminal acute renal failure (ARF) is not widely accepted due to concern about the organ quality. Here we report our initial clinical outcomes of kidney transplantation from DCD donors with ARF. MATERIALS AND METHODS: The results of 29 kidney transplants from ARF DCD donors were compared with those of 60 kidney transplants from non-ARF DCD donors performed at our center from August 2011 to March 2013. RESULTS: There was no difference in the incidence of delayed graft function and acute rejection between ARF and non-ARF kidneys (27.6% vs 16.7%, 10.3% vs 8.3%, respectively). Estimated glomerular filtration rate at 12 months was similar between ARF and non-ARF kidneys. With a mean follow-up of 18 months (range 7 to 26 months), actual patient and graft survival rates for ARF DCD recipients were 100% and 96.6%, respectively, which were similar to those of the control group of kidney transplants from non-ARF kidneys (98.3% and 95.0%). CONCLUSIONS: Kidneys from DCD donors with terminal ARF have excellent short-term outcomes and may represent another potential method to safely expand the donor pool.

Escherichia coli infection induces autoimmune cholangitis and anti-mitochondrial antibodies in non-obese diabetic (NOD).B6 (Idd10/Idd18) mice.

Several epidemiological studies have demonstrated that patients with primary biliary cirrhosis (PBC) have a higher incidence of urinary tract infections (UTI) and there is significant homology of the immunodominant mitochondrial autoantigen, the E2 component of the pyruvate dehydrogenase complex (PDC-E2), between mammals and bacteria. Previous work has demonstrated that non-obese diabetic (NOD).B6 Idd10/Idd18 infected with Novosphingobium aromaticivorans developed liver lesions similar to human PBC. It was postulated that the biliary disease was dependent upon the presence of the unique N. aro glycosphingolipids in activating natural killer T (NK T) cells. To address this issue, we infected NOD.B6 Idd10/Idd18 mice with either Escherichia coli, N. aro or use of a phosphate-buffered saline (PBS) vehicle control and serially followed animals for the appearance of liver pathology and anti-mitochondrial autoantibodies (AMA). Of striking importance, the biliary disease of E. coli-infected mice was more severe than N. Aro-infected mice and the titre of AMA was higher in E. coli-infected mice. Furthermore, the immunopathology did not correlate with the ability of bacterial extracts to produce antigen-dependent activation of NK T cells. Our data suggest that the unique glycosphingolipids of N. aro are not required for the development of autoimmune cholangitis. Importantly, the data highlight the clinical significance of E. coli infection in a genetically susceptible host, and we suggest that the appearance of autoimmune cholangitis is dependent upon molecular mimicry. These data highlight that breach of tolerance to PDC-E2 is probably the first event in the natural history of PBC in genetically susceptible hosts.

Human intrahepatic biliary epithelial cells engulf blebs from their apoptotic peers.

The phagocytic clearance of apoptotic cells is critical for tissue homeostasis; a number of non-professional phagocytic cells, including epithelial cells, can both take up and process apoptotic bodies, including the release of anti-inflammatory mediators. These observations are particularly important in the case of human intrahepatic biliary cells (HiBEC), because such cells are themselves a target of destruction in primary biliary cirrhosis, the human autoimmune disease. To address the apoptotic ability of HiBECs, we have focused on their ability to phagocytize apoptotic blebs from autologous HiBECs. In this study we report that HiBEC cells demonstrate phagocytic function from autologous HiBEC peers accompanied by up-regulation of the chemokines CCL2 [monocyte chemotactic protein-1 (MCP-1)] and CXCL8 [interleukin (IL)-8]. In particular, HiBEC cells express the phagocytosis-related receptor phosphatidylserine receptors (PSR), implying that HiBECs function through the 'eat-me' signal phosphatidylserine expressed by apoptotic cells. Indeed, although HiBEC cells acquire antigen-presenting cell (APC) function, they do not change the expression of classic APC function surface markers after engulfment of blebs, both with and without the presence of Toll-like receptor (TLR) stimulation. These results are important not only for understanding of the normal physiological function of HiBECs, but also explain the inflammatory potential and reduced clearance of HiBEC cells following the inflammatory cascade in primary biliary cirrhosis.

Kidney transplantation from donors after cardiac death: an initial report of 71 cases from China.

Shortage of deceased donors is a severe problem in recent years in China especially in a culture in which brain death criteria is not widely accepted. Donation after cardiac death (DCD) has been reported to expand the donor pool despite higher rates of primary nonfunction (PNF) and delayed graft function (DGF) after transplantation. We collected 71 DCD kidney transplants performed at our hospital between February, 2007 and June, 2012 with aims to demonstrate the feasibility of DCD donation in China. All patients were followed up, and postoperative complications and graft loss were recorded. The PNF rate was 2.8%, and DGF rate was 28.2%. The 1- and 3-year graft survival was 95.7% and 92.4%. In conclusion, graft survival of DCD kidney transplantation in China is excellent despite of higher rates of PNF and DGF after transplantation.