[Clinicopathological features of gastric carcinomas with NTRK-rearrangement/amplification: report of four cases].

Objective: To investigate the clinicopathological, immunohistochemical and molecular genetic characteristics of gastric carcinoma with NTRK-rearrangement/amplification. Methods: The clinicopathological data of gastric carcinoma cases with NTRK-rearrangement/amplification diagnosed from January 2011 to September 2020 at the Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, China, were collected. The clinicopathological, immunophenotypic and molecular pathological features were analyzed. The relevant literature was reviewed. Results: There were 4 cases of gastric carcinoma with NTRK-rearrangement/amplification. All 4 patients were male, aged 57-67 years (average, 63 years). Tumor sizes ranged from 3.5 to 5.2 cm (average, 4.8 cm). All tumors were in the antrum. All 4 patients underwent radical gastrectomy and were followed up after the surgery. Morphologically, all tumors showed histological features with enteroblastic-differentiated gastric carcinoma. Tumor cells showed predominantly tubular/papillary architecture, with conspicuous vesicular nuclei and pale staining or transparent cytoplasm. Immunohistochemistry showed pan-TRK expression in all cases, with various degrees of positivity in the cytoplasm. All cases were subject to NTRK1/2/3 detection using fluorescence in situ hybridization. There were NTRK translocations in 2 cases and NTRK amplifications in 2 cases. These cases were further verified by RNAseq next generation sequencing which confirmed that NTRK1 gene translocation (TPM3-NTRK1) and NTRK2 gene translocation (NTRK2-SMCHD1) occurred in two cases, respectively. Conclusions: NTRK mutation occurs less frequently in gastric cancer. In this study, the cases mainly occur in the antrum. The morphology has the characteristics of enteroblastic differentiation. The tumors have unique histological, immunophenotypic and molecular characteristics, which require much attention from pathologists to effectively guide clinicians to choose the best treatment.

[Correlation of NTRK genetic fusions with mismatch repair protein deletion in patients with colorectal cancer].

Objective: To investigate the relationship between the expression of four mismatch repair proteins (MLH1, MSH2, MSH6 and PMS2) and NTRK genetic fusions in colorectal cancer. Methods: The paraffin-embedded tissue blocks of 830 cases of colorectal cancer were collected at the Affiliated Drum Tower Hospital, Nanjing University Medical School, China, from 2015 to 2019. Immunohistochemical and fluorescence in situ hybridization(FISH) method were used respectively to detect the expression of mismatch repair proteins and the break-apart of NTRKs; and the relationship between the expression of mismatch repair proteins and the NTRK genetic fusions was analyzed. Results: The overall mismatch repair protein deficiency (dMMR) rate was 9.88% (82/830), the mismatch repair proteins proficiency (pMMR) rate was 90.12%(748/830). The total deficiency rate of MLH1 protein was 9.04% (75/830), hPMS2 protein deficiency rate was 9.04% (75/830), MSH2 protein deficiency rate was 2.53% (21/830), MSH6 protein deficiency rate was 4.10% (34/830), the deficiency rate of synchronous MLH1 and PMS2 were 8.67% (72/830) and the deficiency rate of synchronous MSH2 and MSH6 were 2.17% (18/830). The dMMR group was associated with tumor location, different histological subgroups, tumor differentiation, AJCC stage and N stage (P<0.05). There were six cases (7.32%) carrying NTRK fusion by FISH among the 82 cases of dMMR, but only seven cases (0.94%) carrying NTRK fusion among the 748 cases of PMMR. The NTRKs translocation by FISH in all 13 cases were further confirmed by next generation sequencing. Among the clinicopathological characteristics, only differentiation showed significant difference between NTRK fusion positive and negative groups (P<0.05). More importantly, NTRK fusion was enriched in dMMR group (7.32% vs. 0.94%). Conclusion: In dMMR colorectal cancer group, the prevalence of NTRK fusion is higher than that in pMMR group.

[Clinical features of children with cow's milk sensitization].

Objective: To investigate sensitization rate of cow's milk in children, and explore its clinical features. Methods: This study enrolled a total of 818 patients under 18 years old with suspected food allergy who were admitted to the Allergy department in Beijing Shijitan Hospital during June 2018 to November 2020. The ImmunoCAP fluorescent enzyme-linked immunoassay system was used to quantify cow milk-specific immunoglobulin E (sIgE). Mild sensitization to cow's milk was defined as Radio-Allergo-Sorbent-Test (RAST) class 1, moderate sensitization was defined as class 2-3 and severe sensitization was class 4-6. Statistical methods such as χ2 test, independent sample t-test, one-way analysis of variance, and Spearman correlation analysis were used to retrospectively clarify differences of cow's milk sensitization rate between ages of children and elaborate its clinical features. Results: Overall sensitization rate of cow's milk reached 25.7% (210/818). Positive rate of cow milk sensitization (39.2%), cow milk sIgE levels [0.93 (0.52, 2.62)] kU/L, and moderate to severe sensitization rate (23.5%) were highest in infants aged between 0-3 years old. The sensitization rate and severity of sensitization declined with age. Most common clinical manifestation of cow milk sensitization was skin symptoms (50.0%), followed by respiratory symptoms (38.9%) and gastrointestinal symptoms (36.1%). Skin symptoms were the most common manifestation in 0-3 year-old group (47.3%), and respiratory symptoms were more common in 4-6 and 7-18 year-old groups (58.7%, 56.0%). Multiple-sensitization rate of patients with moderate to severe cow milk sensitization was 74.1%, most of which (70.4%) were co-sensitized by other food allergens, and 31.5% were co-sensitized by inhaled allergens. Conclusions: In population with age under 18 years old, infants aged between 0-3 years old suffered highest cow milk sensitization rate and increased sensitization severity. Then the severity decreased with age increasing. Patients with cow milk sensitization manifested skin symptoms most.

[Study on the safety and clinical efficacy of osteotomy after halo pelvic traction in severe scoliosis accompanied with split cord malformation].

Objective: To investigate the safety and clinical efficacy of osteotomy after halo pelvic traction in severe scoliosis accompanied with split cord malformation. Methods: The clinical data of 14 patients with severe scoliosis accompanied with split cord malformation admitted to the Department of Spinal Surgery, Guizhou Orthopedic Hospital from August 2015 to August 2019 were retrospectively analyzed.There were 6 males and 8 females, aged (19.8±5.0) years (range:13 to 34 years). All patients received spinal orthopedic surgery after halo pelvic traction for 3 to 7 weeks.The data of traction time, height, Cobb angle in the main curved coronal plane and sagittal plane, lung function and nutritional status of the patient were collected before and after the treatment. Paired t test was used to compare the evaluation indexes. Results: The traction time of the 14 patients was (35.2±8.3)days (range:20 to 49 days), and the height of them increased from (156.7±7.6)cm (range:141 to 166 cm) before traction to (167.0±6.4)cm (range:154 to 177 cm) after traction（t=-10.49，P<0.01）. The Cobb angle on the main curved coronal plane decreased from (117.4±17.2) ° (range: 91°to 176°) before traction to (56.4±8.1) ° (range:44°to 68°) after traction（t=13.90，P<0.01）. The sagittal Cobb angle decreased from (92.5±11.6) ° (range:62°to 132°) before traction to (41.7±7.7) °(range:29°to 51°) after traction（t=12.11，P<0.01）. No complications such as loosening of nailing and infection occurred during traction, and no decrease of nerve function occurred. Nine patients underwent single segment acromial transpedicle osteotomy and five underwent double segment adjacent asymmetric shortening osteotomy. None of the patients underwent longitudinal fracture resection. The lung function and nutritional status were improved after traction and surgery(all P<0.01). Postoperative follow-up was (22.5±9.1)months (range:12 to 36 months). At the last follow-up, the coronal Cobb angle was (56.3±7.1) °, and the sagittal Cobb angle was (37.7±6.5) °, showing no statistically significant difference from the angle after traction(t=0.16,P=0.88; t=2.28,P=0.32). There was no loss of orthopedic angle. None of the patients had internal fixation displacement, loosening or fracture. Conclusion: The treatment of severe scoliosis with accompanied with split cord malformation by halo pelvic traction is safe and effective, which is worthy of further confirmation by large sample study.

[Current situation of screening, prevention and treatment of bleeding esophageal varices in cirrhotic portal hypertension in Tibet region: a multicenter study].

Objective: To investigate and analyze the current situation, screening, clinical characteristics, prevention and treatment of bleeding esophageal varices in cirrhotic patients with portal hypertension in Tibet region. Methods: Clinical data of cirrhotic patients with portal hypertension through March 2017 to February 2020 from Tibet region were collected and analyzed retrospectively. Results: 511 cases with liver cirrhosis were included in the study, of which 185 cases (36.20%) had compensated cirrhosis and 326 cases (63.80%) had decompensated cirrhosis. Further analysis of the etiological data of liver cirrhosis showed that 306 cases (59.88%) were of chronic hepatitis B, 113 cases (22.11%) of alcoholic liver disease, and 68 cases (13.31%) of chronic hepatitis B combined with alcoholic liver disease. Among patients with compensated liver cirrhosis, 48 cases (25.95%) underwent endoscopic examination of which 33 diagnosed as high-risk variceal bleeding. However, none of these 33 cases had received non-selective ß-blocker therapy, and only four patients had received endoscopic variceal banding therapy. Among patients with decompensated liver cirrhosis, 83 cases (25.46%) had a history of upper gastrointestinal bleeding, 297 cases (91.10%) had ascites, 23 cases (7.05%) had hepatic encephalopathy, and 3 cases (0.92%) had hepatorenal syndrome. Among the patients with a history of upper gastrointestinal bleeding, 42 cases (50.60%) had received secondary preventive treatment for bleeding esophageal varices, including 39 cases of endoscopic treatment, 1 case of endoscopic combined drug treatment, 3 cases of interventional treatment, and 2 cases of surgical treatment. Conclusion: Chronic hepatitis B and alcoholic liver diseases are the main causes of liver cirrhosis in Tibet region. Moreover, this region lacks screening, prevention and treatment for bleeding esophageal varices in cirrhotic patients with portal hypertension. Therefore, it is necessary to increase the screening of high-risk groups to prevent and improve the first-time bleeding, and promote multidisciplinary team to prevent and treat re-bleeding.

[Expression of SOX-11 and TFE3 in solid-pseudopapillary tumor of pancreas and its clinical significance].

Objective: To investigate the expression status and diagnostic value of SRY related high mobility group box 11 (SOX-11) and transcription factor E-3 (TFE3) in solid pseudopapillary tumors of pancreas (SPTPs). Methods: Thirty-eight cases of SPTPs, 36 cases of well-differentiated pancreatic neuroendocrine tumors (PanNETs) and six cases of pancreatic acinar cell carcinomas (PACCs) were collected at the Affiliated Drum Tower Hospital of Nanjing University Medical School from 2012 to 2019. The expression of SOX-11, TFE3 and ß-catenin was detected by immunohistochemistry, and the TFE3 gene status was detected by FISH in 18 cases of SPTPs. Results: Among the 38 SPTP patients, 29 were female and 9 were male, with a mean age of 50 years; among 36 PanNET patients, 32 were female and 4 were male, with a mean age of 39 years; for the six PACC patients, four were male and two were female, with a mean age of 60 years. ß-catenin was positive in all 38 SPTPs, but was negative in all 36 PanNETs and 5/6 PACCs. SOX-11 was positive in 35/38 (92.1%) of SPTPs, but was negative in all 36 PanNETs and 6 PACCs. TFE3 was positive in 36/38 (94.7%) of SPTPs, but was negative in all 36 PanNETs and 6 PACCs. Among these three tumors, the specificity and sensitivity of ß-catenin were 97.6% and 100.0%, the specificity and sensitivity of SOX-11 were 92.1% and 100.0%, the specificity and sensitivity of TFE3 were 94.7% and 100.0%, respectively. There was a significant difference of the expression status of all three markers in SPTPs compared with PanNETs and PACCs (P<0.01). The results of SOX-11 and TFE3 immunostaining showed high consistency (Kappa>0.6). No gene rearrangement (0/18) of TFE3 was found in SPTPs. Conclusion: SOX-11 and TFE3 are highly expressed in SPTPs, and their specificity in the differential diagnosis of SPTPs is better than that of ß-catenin.

Long noncoding RNA CRNDE promotes proliferation, migration and invasion in prostate cancer through miR-101/Rap1A.

Prostate cancer (Pca) is the second frequent malignancy in men. Long noncoding RNAs (lncRNAs) have been reported to play essential roles in the progression of cancers, including Pca. LncRNA colorectal neoplasia differentially expressed (CRNDE) has been found to affect tumorigenesis in many cancers. However, the exact role and mechanism of CRNDE in Pca remain poorly understood. 64 Pca patients were recruited in this study. PC3 and 22RV1 cells were used in vitro experiments. The expressions of CRNDE, microRNA-101 (miR-101), and Ras-related protein 1A (Rap1A) were detected in vivo and in vitro by quantitative real-time polymerase chain reaction and western blot, respectively. Cell proliferation, apoptosis, migration, and invasion were investigated through cell counting kit-8, flow cytometry, and Transwell assays, respectively. The interaction between miR-101 and CRNDE or Rap1A was explored by bioinformatics analysis and luciferase reporter assay. High expression of CRNDE was shown in Pca tissues and cells and predicted poor outcomes of patients. Overexpression of CRNDE promoted cell proliferation, migration, and invasion but decreased apoptosis in Pca cells, while its knockdown showed an opposite effect. CRNDE was a decoy of miR-101 and its effect on Pca progression was reversed by miR-101. Rap1A was identified as a target of miR-101 and it attenuated the effect of miR-101 on Pca development. Moreover, the Rap1A protein level was positively regulated by CRNDE, which was weakened by miR-101. CRNDE contributed to cell proliferation, migration, and invasion by regulating the miR-101/Rap1A axis in Pca, providing a novel strategy for Pca treatment.

Recovery of stem cell proliferation by low intensity vibration under simulated microgravity requires LINC complex.

Mesenchymal stem cells (MSC) rely on their ability to integrate physical and spatial signals at load bearing sites to replace and renew musculoskeletal tissues. Designed to mimic unloading experienced during spaceflight, preclinical unloading and simulated microgravity models show that alteration of gravitational loading limits proliferative activity of stem cells. Emerging evidence indicates that this loss of proliferation may be linked to loss of cellular cytoskeleton and contractility. Low intensity vibration (LIV) is an exercise mimetic that promotes proliferation and differentiation of MSCs by enhancing cell structure. Here, we asked whether application of LIV could restore the reduced proliferative capacity seen in MSCs that are subjected to simulated microgravity. We found that simulated microgravity (sMG) decreased cell proliferation and simultaneously compromised cell structure. These changes included increased nuclear height, disorganized apical F-actin structure, reduced expression, and protein levels of nuclear lamina elements LaminA/C LaminB1 as well as linker of nucleoskeleton and cytoskeleton (LINC) complex elements Sun-2 and Nesprin-2. Application of LIV restored cell proliferation and nuclear proteins LaminA/C and Sun-2. An intact LINC function was required for LIV effect; disabling LINC functionality via co-depletion of Sun-1, and Sun-2 prevented rescue of cell proliferation by LIV. Our findings show that sMG alters nuclear structure and leads to decreased cell proliferation, but does not diminish LINC complex mediated mechanosensitivity, suggesting LIV as a potential candidate to combat sMG-induced proliferation loss.

[Clinical characteristics of the Wilson disease carrier].

Objective: To study the clinical symptoms, copper metabolism and imaging characteristics of Wilson disease (WD) carriers and to explore the treatment strategy of WD carriers. Methods: Forty WD carriers, 40 WD patients and 20 normal controls from the First Affiliated Hospital of Sun Yat-sen University from July 2007 to May 2018 were included. The modified Young scale was used for neural symptom scoring, and Child grading of liver function, mental symptoms rating scale, magnetic resonance imaging (MRI) scan, susceptibility weighted imaging (SWI) inspection, metal metabolism tests were also applied to all the included subjects. Corrected phase (CP) was measured by SWI. WD carriers were divided into symptomatic group and asymptomatic group. Symptomatic WD carriers were treated with penicillamine for 2 weeks and zinc gluconate for 3 months, then their neurological symptoms, liver function grade, metal metabolism index were rechecked. Results: Six WD carriers presented with some clinical symptoms, including 5 with neurological symptoms and 4 with liver dysfunction. The score of Hamilton anxiety (HAMA) scale of symptomatic WD carriers was higher than that of normal control group (P=0.021). 85% of carriers had ceruloplasmin level less than 0.26 g/L. 80% of carriers had serum copper between normal controls and WD patients. The free copper level of WD carriers was lower than that of WD patients (P=0.012, 0.019). Urinary copper in symptomatic WD carriers was higher than normal controls (P=0.047). The CP values of thalamus, globus pallidus and putamen in symptomatic WD carriers were lower than those in normal control group. After treatment with penicillamine in symptomatic WD carriers, urinary copper was higher than that before treatment (P=0.036). After treatment, the liver enzymes of symptomatic WD carriers returned to normal, and the score of modified Young scale was lower than before treatment (P=0.031). Conclusions: Mild copper metabolism abnormality is seen in WD carriers. A few carriers have neurological symptoms such as limb tremors, or liver symptoms such as abnormal liver enzymes. Abnormal copper metabolism is more serious in symptomatic WD carriers than in asymptomatic WD carriers. Symptomatic WD carriers can be treated with zinc gluconate.

[Transforming growth factor beta regulates hepatic progenitor cells migration via PI3K/AKT/mTOR/p70S6K pathway].

Objective: To investigate the effect and mechanism of transforming growth factor ß (TGFß) on the migration ability of hepatic progenitor cells in vitro. Methods: Primary hepatic progenitor cells of male wild-type C57BL/6J mice were isolated by two-step perfusion method and stimulated with different concentrations of TGFß .The morphological changes were observed under phase -contrast microscopy. The effects of TGFß on migration ability of hepatic progenitor cells were evaluated by scratch test and transwell method. Expression profiling and signaling phospho antibody array detected the signaling pathways involved in the regulation of TGFß on hepatic progenitor cells. Protein level of PI3K/AKT/mTOR/p70S6K signaling pathway and the localization of each signaling molecules in hepatic progenitor cells were detected. Data comparison between the two groups was performed by independent sample t-test. One-way ANOVA was used for data comparison between multiple groups. Results: TGFß made the liver progenitor cells from oval to long spindle type. Scratch test showed that the scratch healing rates of 24 h control group, and 2 ng/ml and 10 ng/ml TGF-beta groups were 36.48% ± 4.37%, 57.35% ± 4.60%, and 73.14% ± 5.02% (F = 65.87, P < 0.01), respectively. Transwell test showed that the number of migrating cells in 24 h control group, 2 ng/ml and 10 ng/ml TGF-beta groups were 127 ± 16, 230 ± 18, and 385 ±36 (F = 94.99, P < 0.01), respectively. The results of expression profiling showed that TGFß regulates gene expression in hepatic progenitor cells, and differentially expressed genes participate in the PI3K-AKT signaling pathway. Signaling phospho antibody array and western blot showed that TGFß regulated PI3K/AKT/mTOR/p70S6K signaling pathway in hepatic progenitor cells. Concurrently, immunofluorescence assay showed phosphorylation (p) 70s6k, p AKT1 and PI3K and F-actin co-localizations. Conclusion: TGFß can promote hepatic progenitor cell migration through PI3K/AKT/mTOR/p70S6K pathway, and p70S6K, pAKT1 and PI3K signaling molecules are involved in the regulation of morphology and migration of liver progenitor cells.

[Effect of emergency thoracic endovascular aortic repair in patients with acute traumatic thoracic aortic injury].

Objective: To investigate the effect of emergency thoracic endovascular aortic repair (TEVAR) in patients with acute traumatic thoracic aortic injury. Method: From January 2014 to December 2016, a total of 35 patients with acute traumatic thoracic aortic injuries were treated with emergency TEVAR in our hospital, their clinical data were analyzed retrospectively in this study. Results: The patients were 42 (34, 55) years old,and there were 31 males.All cases were diagnosed by emergency aorta computed tomography angiography (CTA),and 5 cases were diagnosed as aortic transaction, 13 cases were diagnosed as aortic pseudoaneurysm, 7 cases were diagnosed as aortic dissection, and 10 cases were diagnosed as aortic intramural hematoma combined hemothorax.The concomitant injuries included cerebral contusion (3 cases, 8.6%), pulmonary contusion with rib fracture (31 cases, 88.5%), long bone fracture (7 cases, 22.5%), contusion of viscera or internal organs (3 cases, 8.6%).Emergency TEVAR were performed with vascular suture system preset under local anesthesia after diagnosis,and combined injury was treated in related departments.CTA was repeated after 1, 3 and 6 months and yearly thereafter. One patient died before transferring to catheter room,and 34 (97.1%) patients underwent TEVAR procedure successfully.Time from door to operating room was (88.6±26.6) minutes,and the procedure time was (52.0±9.4) minutes. A total of 69 Perclose Proglide vascular suture system were used,and 2 cases underwent surgical suture because of hematoma and pseudoaneurysm formation in femoral arteries.The involved length of thoracic aorta was (44.5±7.4)mm. A total of 46 stent-grafts were implanted, the length of stent-graft was (164.3±15.2)mm,and the proximal oversize rate was (22.3±8.6)%. The follow-up time was 24 (12,24) months, and there were no procedure related complication such as endoleak and paraplegia. Complete aortic remodeling was observed in 14 cases. Fully thrombolization at stent segments were observed in 7 cases. Fully thrombalization of pseudoaneurysms were observed in 13 cases. One patient complained mild left upper limb weakness due to left subclavian artery occlusion. Conclusion: Emergency TEVAR is safe and effective procedure for the treatment of patients with acute traumatic thoracic aortic injury.

[Clinical analysis of the specific reference intervals of thyroid index for normal pregnant women].

Objective: To explore the reference intervals of thyrotropin (TSH) and free thyroxine (FT(4)) in normal pregnant women. Methods: Prospective and longitudinal sequential collection of the cases were performed. A total of 155 singleton pregnant women who had regular prenatal examination and delivery in West China Second Hospital of Sichuan University from January 2015 to December 2015 were included as the research group. Blood samples were sequentially collected from the pregnant women at the first (9-13(+6) weeks) , the second (24-27(+6) weeks) , the third (32-36 weeks) trimesters, respectively. Another 155 non-pregnant women were selected as the control group simultaneously, in an effort to establish a range of reference intervals of thyroid index in each period of pregnancy. Meanwhile, neonatal plantar blood were also collected on special filter paper and TSH levels were measured. Results: (1) TSH reference intervals [percentile 2.5-percentile 97.5 (P(2.5)-P(97.5)) ] were 0.08-3.29 mU/L, 0.59-4.22 mU/L and 0.81-4.33 mU/L in three trimesters respectively. FT(4) reference intervals were 11.88-20.06 pmol/L, 9.89-15.80 pmol/L and 9.22-15.77 pmol/L in three trimesters respectively. (2) The median serum TSH in the first trimester (1.35 mU/L) was lower than that in the second trimester (2.15 mU/L) , the third trimester (2.19 mU/L) , and the control group (2.19 mU/L) . The differences were all statistically significant (P<0.05) . There was no significant difference in median serum TSH between the second trimester and the third trimester, the second trimester and the control group, and the third trimester versus the control group. (3) The median serum FT(4) in the first trimester (15.16 pmol/L) was higher than that in the second trimester (12.39 pmol/L) and the third trimester (12.26 pmol/L) . The differences were both statistically significant (P<0.05) . The median FT(4) in the second trimester and the third trimester was lower than that in the control group (15.64 pmol/L) , and the differences were both statistically significant (P<0.05) . (4) In the first trimester, the median serum total triiodothyronine (TT(3)) level (2.32 nmol/L) was higher than that in the control group (1.56 nmol/L) , total thyroxine (TT(4)) level (154.60 nmol/L) was also higher than that in the control group (98.25 nmol/L) , and free triiodothyronine (FT(3)) level (4.70 pmol/L) was lower than that in the control group (4.84 pmol/L) , the differences were all statistically significant (P<0.05) . (5) The TSH levels of all neonatus were normal [ (2.1±1.3) mU/L]. Conclusions: The thyroid hormone levels between pregnant women and non-pregnant women are significantly different. Moreover, the reference intervals are vary with pregnancy period. It is important to establish the specific reference intervals of thyroid hormones in the first, second and third trimester of pregnancy specific to local region or unit.

Immunoproteomics to identify Staphylococcus aureus antigens expressed in bovine milk during mastitis.

Staphylococcus aureus is an opportunistic pathogen affecting both human and animal species. An effective vaccine to prevent S. aureus bovine disease and transmission would have positive effects on animal well-being, food production, and human health. The objective of this study was to identify multiple antigens that are immunoreactive during udder colonization and disease for exploration as vaccine antigens to prevent bovine mastitis. Staphylococcus aureus produces several cell wall-anchored and surface-associated virulence factors that play key roles in the pathogenesis of mastitis. Many of these proteins are conserved between different strains of S. aureus and represent promising vaccine candidates. We used an immunoproteomics approach to identify antigenic proteins from the surface of S. aureus. The expression of cell wall and surface proteins from S. aureus was induced under low iron conditions, followed by trypsin extraction and separation by 2-dimensional electrophoresis. The separated proteins were blotted with antibodies from mastitic bovine milk and identified by liquid chromatography-mass spectrometry. Thirty-eight unique proteins were identified, of which 8 were predicted to be surface exposed and involved in S. aureus virulence. Two surface proteins, iron-regulated surface determinant protein C (IsdC) and ESAT-6 secretion system extracellular protein (EsxA), were cloned, expressed, and purified from Escherichia coli for confirmation of immune reactivity by ELISA. A PCR of 37 bovine S. aureus isolates indicated that the presence of esxA and isdC is conserved, and amino acid alignments revealed that IsdC and EsxA sequences are highly conserved. The immunoproteomics technique used in this study generated reproducible results and identified surface exposed and reactive antigens for further characterization.

[Feasibility and efficacy of transcatheter closure of anastomotic leakage after aortic surgery using Amplatzer Vascular Plug â ¢].

Objective: To investigate the feasibility and efficacy of transcatheter closure of anastomotic leakage after aortic surgery using Amplatzer Vascular Plug Ⅲ (AVP Ⅲ). Methods: A retrospective analysis was performed in 5 patients with anastomotic leakage after aortic surgery, who underwent transcatheter closure in our hospital from January to June 2017 using AVP Ⅲ. Surgeries were performed in 3 cases of Standford type A dissection, 1 case of ascending aortic aneurysm and 1 case of persistent truncus.There were 3 males,and age was (43.8±13.1) years old. Anastomotic leakages located at the ascending aorta in 4 patients, and the other one located between the aortic arch and the stent-graft.Three of them had aorta-right atrium fistula and patients suffered from progressive heart failure. False aneurysm between aorta and pulmonary artery was formed in 1 patient, and patent aortic false lumenwas found in the other patient. All the AVP Ⅲ were deployed based on a femoral arteriosus loop. Patients were followed up after transcatheter closure to observe the clinical results. Results: Six AVP Ⅲ were successfully implanted in the 5 patients. Trivial residual shunt was seen in 1 patient after closure. The patients were followed up 6 (1, 6) months. The cardiac function improved from NYHA class Ⅱ-Ⅳ to class Ⅰ-Ⅱ after the procedure in 3 congestive heart failure patients.The right atrium systolic pressure was significantly reduced after the procedure((8.7±1.8) mmHg (1 mmHg=0.133 kPa) vs. (24.3±2.3) mmHg, P=0.03). The diameter of the false aneurysm reduced in 1 patient after the procedure. Complete thrombosis formation of the thoracic false lumen was observed in 1 patient. Conclusion: Transcatheter closure of anastomotic leakage after aortic surgery using AVP Ⅲ is feasible and effective according to our primary experience.