[A case of fungal conjunctivitis manifested by a conjunctival mass].

A 71-year-old man presented with a history of persistent redness and swelling in the left eye accompanied by an enlarging mass in the conjunctiva. He denied any history of trauma. Local anti-inflammatory treatment was ineffective. Slit lamp examination demonstrated a tough and immobile grayish broad basal mass at the corneal limbus and bulbar conjunctiva and a local bulge of 3 mm × 2 mm at the medial and lateral side of the upper palpebral conjunctiva near the eyelid margin. The excisional biopsy showed granulomatous inflammation with irregular and atypical squamous epithelial hyperplasia. Histopathology and immunohistochemical staining revealed a fungal infection. The secretion smear examination was performed to clarify the pathogen as Candida albicans, and chronic fungal maxillary sinusitis was found through imaging tests. Thus a diagnosis of conjunctival candidiasis was made. The conjunctival mass subsided after systemic and local antifungal therapy.

[Sepsis and acute kidney injury].

Sepsis is a life-threatening multiple organ dysfunction syndrome caused by infection, and kidney is one of the most organs susceptible to injury in sepsis. Critical ill patients with sepsis often suffer from acute kidney injury (AKI) of varying degrees with a high mortality. There are more and more researches on the early identification, pathogenesis, diagnosis and treatment of acute kidney injury in sepsis. In order to improve the understanding and reduce the mortality of acute kidney injury, this review briefly discussed the early identification, physio-pathologic mechanism, treatment, prognosis and follow up.

Predictive value of cell cycle arrest biomarkers for cardiac surgery-associated acute kidney injury: a meta-analysis.

BACKGROUND: A biomarker test based on a combination of urine tissue inhibitor of metalloproteinases 2 (TIMP-2) and insulin-like growth factor binding protein 7 (IGFBP7) has been used as a potential biomarker of acute kidney injury (AKI). This meta-analysis aimed to evaluate the predictive value of this biomarker for cardiac surgery-associated acute kidney injury (CSA-AKI). METHODS: We searched MEDLINE, PubMed, Cochrane, and EMBASE for studies. We evaluated the methodological quality of each included study using the Quality Assessment of Diagnostic Accuracy Studies 2 criteria. Meta-DiSc and STATA were used for statistical analyses. RESULTS: A total of 10 studies (747 patients) were included in this meta-analysis. Pooled sensitivity and specificity with corresponding 95% confidence intervals (CI) were 0.77 (95% CI: 0.70-0.83, I2=40.7%) and 0.76 (95% CI: 0.72-0.79, I2=69.1%), respectively. Pooled positive likelihood ratio (LR), negative LR, and diagnostic odds ratio were 3.26 (95% CI: 2.51-4.23, I2=50.7%), 0.32 (95% CI: 0.24-0.41, I2=6.7%), and 10.08 (95% CI: 6.85-14.84, I2=6.7%), respectively. The area under the curve estimated by summary receiver operating characteristics was 0.83 [standard error (SE) 0.023] with a Q* value of 0.759 (se 0.021). There was no heterogeneity amongst the 10 studies from both threshold and non-threshold effects. Subgroup analysis showed that the diagnostic value was related to the severity of AKI and time measurement. CONCLUSIONS: Urinary [TIMP-2]·[IGFBP7] is an effective predictive test for cardiac surgery associated acute kidney injury with good diagnostic accuracy within 24 h. Studies examining use of biomarker-guided care bundles are indicated.

Simultaneous construction of PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps.

UNLABELLED: The aim of this study was to simultaneously construct PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps at 4 and 10°C. Calibration curves were established to correlate peak density of DGGE bands with microbial counts. Microbial counts derived from PCR-DGGE and plate methods were fitted by Baranyi model to obtain molecular and traditional predictive models. For L. monocytogenes, growing at 4 and 10°C, molecular predictive models were constructed. It showed good evaluations of correlation coefficients (R(2) > 0.92), bias factors (Bf ) and accuracy factors (Af ) (1.0 ≤ Bf ≤ Af ≤ 1.1). Moreover, no significant difference was found between molecular and traditional predictive models when analysed on lag phase (λ), maximum growth rate (µmax ) and growth data (P > 0.05). But for V. parahaemolyticus, inactivated at 4 and 10°C, molecular models show significant difference when compared with traditional models. Taken together, these results suggest that PCR-DGGE based on DNA can be used to construct growth models, but it is inappropriate for inactivation models yet. This is the first report of developing PCR-DGGE to simultaneously construct multiple molecular models. SIGNIFICANCE AND IMPACT OF THE STUDY: It has been known for a long time that microbial predictive models based on traditional plate methods are time-consuming and labour-intensive. Denaturing gradient gel electrophoresis (DGGE) has been widely used as a semiquantitative method to describe complex microbial community. In our study, we developed DGGE to quantify bacterial counts and simultaneously established two molecular predictive models to describe the growth and survival of two bacteria (Listeria monocytogenes and Vibrio parahaemolyticus) at 4 and 10°C. We demonstrated that PCR-DGGE could be used to construct growth models. This work provides a new approach to construct molecular predictive models and thereby facilitates predictive microbiology and QMRA (Quantitative Microbial Risk Assessment).

Clinical application of three-dimensional reconstruction and rapid prototyping technology of multislice spiral computed tomography angiography for the repair of ventricular septal defect of tetralogy of Fallot.

Three-dimensional (3D) reconstruction and rapid prototyping technology (RPT) of multislice spiral computed tomography angiography (CTA) was applied to prepare physical models of the heart and ventricular septal defects of tetralogy of Fallot (ToF) patients in order to explore their applications in the diagnosis and treatment of this complex heart disease. CTA data of 35 ToF patients were collected to prepare l:l 3D solid models using digital 3D reconstruction and RPT, and the resultant models were used intraoperatively as reference. The operations of all 35 patients were completed under the guidance of the 3D solid model, without difficulty. Intraoperative findings of the patients were consistent with the morphological and size changes of the 3D solid model, and no significant differences were found between the patches obtained from the 3D solid model and the actual intraoperative measurements (t = 0.83, P = 0.412). 3D reconstruction and RPT of multislice spiral CTA can accurately and intuitively reflect the anatomy of ventricular septal defects in ToF patients, providing the foundation for a solid model of the complex congenital heart.

Dephosphorylation of NSSR1 regulates alternative splicing of the GluR-B minigene.

Neural salient serine/arginine-rich protein 1 (NSSR1, alternatively SRp38) is an important splicing factor that can repress pre-mRNA alternative splicing in cells during heat shock and mitosis. We show here that NSSR1 protein is dephosphorylated when cells are heat shocked or incubated with kinase inhibitor K252a. Both heat shock and K252a treatment increase the truncated splicing isoform of the GluR-B minigene pre-mRNA. We also investigated the roles of the RRM motif and three RS domains of NSSR1 in in vivo pre-mRNA splicing. The results show that deletion of the RRM motif did not affect GluR-B minigene pre-mRNA splicing, but deletion of any one of the three RS domains increases the truncated splicing isoform of the GluR-B minigene. We further show that an SRSRSK sequence in the RS3 domain may play an important role in the function of NSSR1 in pre-mRNA splicing.

Meta-analysis of risk factors for restenosis after stent implantation in patients with ischemic cerebrovascular disease.

OBJECTIVE: The aim of this study was to investigate the risk factors for restenosis after stent implantation in patients with ischemic cerebrovascular disease (ICVD), and to provide a reference for potential measures to prevent ICVD. MATERIALS AND METHODS: Relevant studies were identified by searching PubMed, ScienceDirect and Web of Science databases. Combined adjusted odds ratios and 95% confidence intervals were calculated. RESULTS: Seven case-control studies were identified in the end. Diabetes mellitus and residual stenosis were the two main risk factors for restenosis (OR = 0.59, 95% CI: 0.39-0.91, p = 0.01; OR = 36.73, 95% CI: 19.72-70.02, p < 0.001). Gender, smoking, hypertension, dyslipidemia, and stent type were not significantly associated with restenosis (OR = 0.85, 95% CI: 0.53-1.38, p = 0.52; OR = 1.30, 95% CI: 0.91-1.86, p = 0.15; OR = 0.86, 95% CI: 0.16-4.66, p = 0.86; OR = 1.30, 95% CI: 0.58-2.91, p = 0.53; OR = 1.34, 95% CI: 0.72-2.48, p = 0.35). CONCLUSIONS: The prevention of restenosis after stenting is particularly important for ICVD patients with diabetes or a high residual stenosis rate.

Screening of key genes associated with contused rat spinal cord with DNA microarray.

OBJECTIVES: To identify key genes and novel potential therapeutic targets for contused spin cord injury through analyzing microarray data. MATERIALS AND METHODS: Gene expression data set GSE2599 was downloaded from Gene Expression Omnibus, including 3 rat spinal cord injury (SCI) samples and 3 healthy controls. Data pre-treatment and differential analyses were performed with packages of R. Cluster analysis was done with gene expression values to globally present the difference between the two states. Functional enrichment analysis was performed for all the DEGs with DAVID tools. The most up- and down-regulated genes were picked out and their interactors were predicted with String. Pathway enrichment analysis was done with GENECODIS for all the genes in the network. RESULTS: A total of 227 DEGs were screened out, 132 up-regulated genes and 145 down-regulated genes. Response to wounding, response to organic substance and defense response was the top 3 significant functional terms. APOBEC1 was the most up-regulated gene while HPD was the most down-regulated one. Their interactors were obtained and network was constructed. Pathway enrichment analysis revealed that tyrosine metabolism and other metabolism-related pathways were significantly over-represented. CONCLUSIONS: A range of DEGs were revealed in present study, which could deepen the understandings about the mechanisms of SCI and guide future researches on treatment development.

MiR-126 regulated myocardial autophagy on myocardial infarction.

OBJECTIVE: The aim of the study is to detect the effect of miR-126 in autophagy after acute myocardial infarction (AMI). MATERIALS AND METHODS: First, the changes of miR-126 and autophagy in myocardial infarction tissue and normal heart tissue were compared by establishing the myocardial infarction rat model, so as to clarify the relationship between miR-126 and changes of myocardial infarction and autophagy. Second, the regulation of miR-126 on autophagy related protein Beclin-1 and the role in myocardial infarction were studied to clarify whether miR-126 regulates autophagy through Beclin-1 and participates in the occurrence and development of myocardial infarction. Finally, the relationship between plasma miR-126 was observed. RESULTS: MiR-126 expression can regulate Beclin-1 expression and influence the cardiac function after AMI. The results are of great significance to reveal the new mechanism of myocardial infarction. CONCLUSIONS: The down-regulation of miR-126 will lead to the over-activation of myocardial autophagy induced by Beclin-1, which is an autophagy related protein. The plasm expression of miR-126 may be a clinical marker of autophagy after AMI.

[Clinical analysis of Langerhans cell histiocytosis originating in the base of nasal skull].

Objective:To summarize the clinical features, diagnosis and treatment of Langerhans histiocytosis（LCH） which first appeared in the nasal skull base. Method:Ten cases of LCH with nasal and skull base symptoms were analyzed retrospectively. The clinical characteristics of LCH with nasal and skull base symptoms were summarized. The correlation of other systems involved in LCH was analyzed. Result:Among the 10 patients, the youngest was 1 year and 5 months, and the oldest was 8 years, the average age was 3 years. The main imaging manifestations were osteolytic changes and soft tissue invasion. Seven patients were monofocal and three patients were multifocal. For localized lesions, radical resection and follow-up chemotherapy were performed, and conservative treatment was performed for patients with multiple system involvement and obvious systemic symptoms. Eight patients survived, 2 died. Conclusion:LCH occurs frequently in children and has certain clinical characteristics. Single system and single lesion surgery have a better therapeutic effect, and can achieve a greater survival rate with follow-up chemotherapy.

CalliSpheres® drug-eluting beads (DEB) transarterial chemoembolization (TACE) is equally efficient and safe in liver cancer patients with different times of previous conventional TACE treatments: a result from CTILC study.

PURPOSE: To assess the efficacy and safety of drug-eluting beads transarterial chemoembolization (DEB-TACE) in liver cancer patients with different times of previous conventional transarterial chemoembolization (cTACE) treatments. METHODS: 367 liver cancer patients about to receive DEB-TACE treatment were enrolled in this prospective cohort study. All patients were divided into no previous cTACE group (NPC group), 1-2 times previous cTACE group (PC group) and triple or above previous cTACE group (TPC group) according to the times of previous cTACE treatments. RESULTS: There was no difference in complete response (CR) (P = 0.671) and objective response rate (ORR) (P = 0.062) among three groups. Additionally, no difference in overall survival (OS) among groups (P = 0.899) was found. As to liver function, most liver function indexes were deteriorative at 1 week after DEB-TACE operation, but returned to baseline at 1-3 months after DEB-TACE operation in all three groups, while percentage of abnormal total bile acid (TBA) patients was higher in TPC group than NPC and PC groups at 1-3 month post-DEB-TACE (P = 0.018). As for safety profiles, the incidence of pain during DEB-TACE operation was lower in TPC group compared to NPC and PC groups (P = 0.005), while no difference of other adverse events was found during and 1 month post-DEB-TACE treatment among three groups. CONCLUSION: DEB-TACE treatment was equally efficient and tolerated in liver cancer patients with different times of previous cTACE treatments.

Plasma filtration on mediators of thrombotic microangiopathy: an in vitro study.

OBJECTIVES: Sepsis-induced thrombotic microangiopathy is successfully treated by plasma exchange therapy. However, certain putative mediators of thrombotic microangiopathy may not be removed by plasma filtration. METHODS: We conducted an in vitro study to determine whether plasma filtration can remove ultralarge von Willebrand factor (ULvWF) multimers and other mediators. In separate experiments, human umbilical venous endothelial cell (HUVEC) supernatant enriched with ULvWF or human whole blood was passed through a therapeutic plasma exchange (TPE 2000, PRISMA) filter and samples were taken for measurement of ULvWF, vWF ristocetin cofactor, vWF antigen and PAI-1. RESULTS: The sieving coefficients for vWF and PAI-1 were above 0.9. The ULvWF was gradually eliminated, and nearly disappeared after four circulations. CONCLUSION: The TPE 2000 filter can directly remove potential mediators of sepsis-induced thrombotic microangiopathy.

[Clinical practice of transnasal endoscopic operation for retrobulbar lesions].

Objective: To summarize the skill and experience of transnasal endoscopic operation for retrobulbar lesions. Methods: Seven patients aged from 25 to 67 years old diagnosed as retrobulbar lesions who underwent transnasal endoscopic operation in Department of Otorhinolaryngology Head and Neck Surgery, Xiangya Hospital between January 2013 and October 2016 were retrospectively analyzed. Two males and five females were included in this study. Five patients underwent transnasal endoscopic operation via media rectus-inferior rectus space, with the other 2 cases via media rectus-superior rectus space. Results: Total lesion removal was achieved in 6 of 7 patients, while 1 patient underwent subtotal removal of the lesion. The visual acuity and visual field improved in 3 cases. The pathological examination showed hemangioma(5 cases), bone cyst(1 case) and fibroma(1 case). All patients were followed up for 9 months to 4 years without complications such as eye movement disorder or blindness, except for 1 case with preoperatively proptosis occurred postoperatively transient diplopia. There was no recurrence in 6 patients with total lesion removal, and the patient underwent subtotal removal of fibroma did not undertake operation again. Conclusion: Transnasal endoscopic operation for retrobulbar lesions is a minimally invasive, safe and effective operatiiv method, which could be taken via different surgical approaches according to the size and location of the lesion.

Regulation of the Src tyrosine kinase and Syp tyrosine phosphatase by their cellular association.

The specific activity of the Src tyrosine kinase is elevated in human colon carcinoma cells. To identify Src-binding proteins that might upregulate Src activity in these cells, a human colon carcinoma lambda gt11 expression library was screened with purified, 32P-labeled Src. The SH-PTP2 (Syp) tyrosine phosphatase was isolated and shown to associate with Src. In vitro studies demonstrated that: (1) transforming F527 Src phosphorylates Syp, and (2) Syp dephosphorylates Src at Tyr 527. Both events are known to upregulate enzyme activity. Others have shown that overexpression of the receptor tyrosine phosphatase alpha in rat embryo fibroblasts results in Src activation by dephosphorylation of Tyr 527, cell transformation and tumorigenesis. Thus, transmembrane tyrosine phosphatases may be involved in cell transformation exerting at least some of their effects through activation of Src. To the best of our knowledge, this is the first identification of an intracellular tyrosine, phosphatase which may activate Src by a similar mechanism.

The Shp-2 tyrosine phosphatase activates the Src tyrosine kinase by a non-enzymatic mechanism.

Previously, we demonstrated that the Src tyrosine kinase interacts with the Shp-2 tyrosine phosphatase. To determine whether Shp-2 regulates Src kinase activity, we measured Src activity in cells overexpressing wild-type or catalytically-inactive C463S Shp-2. We observed a 2-3-fold increase in the specific activity of Src in both cell types and the increase did not appear to be due to dephosphorylation of Tyr 527 or phosphorylation of Tyr 416 on Src. Conversely, we observed a 2-3-fold decrease in the specific activity of Src when Shp-2 expression was inhibited. Using glutathione S-transferase-fusion proteins, we demonstrated that Shp-2 binds to the SH3 domain of Src. Our findings reveal that the Shp-2 tyrosine phosphatase can regulate the Src tyrosine kinase by a non-enzymatic mechanism. We also found that the phosphatase activity of Shp-2 immunoprecipitates is downregulated in cells transformed by Src or other proteins, and that Shp-2 preferentially associates with the membrane fraction of transformed cells. We suggest that membrane-association of Shp-2 is important for regulating Shp-2 activity.

A minimum folding unit in the ankyrin repeat protein p16(INK4).

The ankyrin repeat is an abundant, 33 residue sequence motif that forms a consecutive beta-hairpin-helix-loop-helix (beta(2)alpha(2)) fold. Most ankyrin repeat proteins consist of four or more complete repeats, which provide stabilizing interactions between adjacent modules. The cyclin-dependent kinase inhibitor and tumor suppressor p16(INK4) (p16) is one of the smallest ankyrin repeat proteins with a known structure. It consists of four complete repeats plus short N and C-terminal flanking regions that are unstructured in solution. On the basis of preliminary proteolysis studies and predictions using a computer algorithm for identifying autonomous folding units, we have identified a fragment consisting of the third and fourth ankyrin repeats of p16, called p16C, that can fold independently, without the rest of the protein. Far-UV circular dichroism studies showed that p16C has a significant level of alpha-helical secondary structure, and two proline substitutions that disrupt the alpha-helical secondary structure in wild-type p16 disrupt the secondary structure in p16C. The thermal denaturation of p16C is cooperative and reversible, with a midpoint of transition at 30. 5(+/-1) degrees C. From urea-induced denaturation studies, the free energy of unfolding for p16C was estimated to be 1.7(+/-0.3) kcal/mol at 20 degrees C. (1)H-(15)N 2D NMR studies suggest that the ankyrin repeats in p16C are likely to fold into a structure similar to that of full-length p16. In order to define the minimum autonomous folding unit in p16, we have further dissected p16C into two complementary peptides, each containing a single ankyrin repeat. These peptides are unstructured in solution. Thus, p16C is the smallest ankyrin repeat module that is known to fold independently and, in general, we believe that the two-ankyrin repeat fold could be the minimum structural unit for all ankyrin repeat proteins. We further discuss the significance of p16C in protein folding and engineering.

Contribution of individual residues to formation of the native-like tertiary topology in the alpha-lactalbumin molten globule.

Molten globules are partially folded forms of proteins that have native-like secondary structure and a compact geometry, but often without rigid, specific side-chain packing. Recently, the molten globule of alpha-lactalbumin (alpha-LA) has been shown to adopt a native-like tertiary topology, mainly localized in the alpha-helical domain. This native-like topology is reflected by the high effective concentration (Ceff) for formation of the 28-111 disulfide bond, which is approximately 10 to 40 times higher than the Ceff for formation of any non-native disulfide bond in the alpha-helical domain. In order to understand the mechanism for formation of the native-like tertiary topology, we substituted alanine for each of the 23 buried residues in the alpha-helical domain of alpha-LA and determined the effect of these substitutions on the Ceff for formation of the 28-111 disulfide bond. Our results indicate that a subset of hydrophobic residues is most important for formation of the native-like topology. These residues form a densely packed core in the three-dimensional structure of alpha-LA. In contrast, the less important residues consist of both hydrophobic and hydrophilic amino acids located at peripheral positions. These results suggest that a relatively small number of hydrophobic residues may be sufficient for specifying the overall structure of a protein during early stages of protein folding.

Different subdomains are most protected from hydrogen exchange in the molten globule and native states of human alpha-lactalbumin.

alpha-Lactalbumin (alpha-LA) is a two-domain, calcium-binding protein that forms one of the best studied molten globules. We present here amide hydrogen exchange studies of the molten globule formed by human alpha-LA at pH 2 and compare these results with a similar study of the native state at pH 6.3. The most persistent structure in the molten globule is localized in the helical domain, consistent with previous results. However, the helices most protected from hydrogen exchange in the molten globule are, in the native state, less protected from exchange than other regions of the protein. The molten globule appears to contain major elements of the native fold, but formation of the fully native state requires stabilization of structure around the calcium-binding site and domain interface.

A model of dynamic side-chain--side-chain interactions in the alpha-lactalbumin molten globule.

Proteins in the molten globule state contain high levels of secondary structure, as well as a rudimentary, nativelike tertiary topology. Thus, the structural similarity between the molten globule and native proteins may have a significant bearing in understanding the protein-folding problem. To explore the nature of side-chain--side-chain interactions in the alpha-lactalbumin (alpha-LA) molten globule, we determined the effective concentration for formation of the 28--111 disulfide bond in 14 double-mutant proteins, each containing two hydrophobic core residues replaced by alanine. We compared our results with those of single-alanine substitutions using the framework of double-mutant cycle analysis and found that, in the majority of cases, the effects of two alanine substitutions are additive. Based on these results, we propose a model of side-chain-side-chain interactions in the alpha-LA molten globule, which takes into consideration the dynamic nature of this partially folded species.