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Extended left ventricular septal myectomy remains the gold standard for the treatment of hypertrophic obstructive cardiomyopathy (HOCM) with refractory symptoms. On the basis of traditional modified transaortic Morrow myectomy, we innovatively performed a minimally invasive, video-assisted single-port thoracotomy through the right infra-axillary region. Our procedure can provide good visualization of the left ventricular outflow tract and hypertrophic ventricular septum for accurate resection. It also ensures optimal exposure of the mitral valve in the presence of complex mitral subvalvular structures.

Accumulating investigations illustrated that miRNA acts as a key regulator in tumor progression, whereas regulatory role of miR-96-5p in lung adenocarcinoma (LUAD) is warranted. Thus, we sought to probe mechanism of miR-96-5p in this disease. Through bioinformatics analysis, miR-96-5p level in normal tissue and LUAD tissue in TCGA database were obtained. Meanwhile, mRNA expression dataset was analyzed to obtain downregulated mRNAs binding to miR-96-5p. qRT-PCR assessed miR-96-5p and ARHGAP6 mRNA in LUAD. Western blot assessed protein level of ARHGAP6 in LUAD. Dual-luciferase reporter gene detection verified targeting relationship of miR-96-5p and ARHGAP6. Biological functional experiments such as CCK-8, colony formation, scratch healing, and Transwell assessed cell proliferation, migration, and invasion. MiR-96-5p was overexpressed, which fostered LUAD cell proliferation, migration, and invasion. ARHGAP6 was downregulated in LUAD and targeted by miR-96-5p. ARHGAP6 upregulation prominently restored promotion of miR-96-5p on cell progression. MiR-96-5p could stimulate LUAD progression through targeting ARHGAP6. This study generates a novel direction and lays a theoretical basis for targeted therapy.

Background Increasing evidence indicates that the immune microenvironment plays a key role in the genesis and progression of colorectal cancer (CRC). This study aimed to establish an immune-related gene (IRG) signature and determine its clinical prognostic value in patients with CRC. Methods The RNA sequencing and associated clinical data of CRC were downloaded from The Cancer Genome Atlas (TCGA) database. We then screened for differentially expressed IRGs by intersecting with IRGs obtained from the Immunology Database and Analysis Portal. Functional enrichment analyses were carried out to determine the potential biological functions and pathways of the IRGs. We also explored the specific molecular mechanisms of the IRGs by constructing regulatory networks. Prognostic IRGs were obtained by LASSO regression analysis, and subsequently, gene models were constructed in the TCGA dataset to confirm the predictive capacity of these IRGs. Finally, we used the TIMER tool to assess the immune properties of prognostic IRGs and correlate them with immune cells. Results We identified 409 differentially expressed IRGs in patients with CRC. Kyoto Encyclopaedia of Genes and Genomes and Gene Ontology enrichment analyses suggested that these differentially expressed IRGs were significantly related to 102 cancer signalling pathways and various biological functions. Based on the prediction and interaction results, we obtained 59 TF–IRG, 48 miRNA–IRG, and 214 drug–IRG interaction networks for CRC. Four prognostic genes (POMC, TNFRSF19, FGF2, and SCG2) were developed by integrating 47 survival-related IRGs and 42 characteristic CRC genes. The results of gene model showed that patients in the low risk group had better survival outcomes compared to those in the high risk group. The expression of POMC, TNFRSF19, FGF2, and SCG2 was significantly correlated with immune cells. Conclusion This study identified some valid IRGs, and these findings can provide strong evidence for precision immunotherapy in patients with CRC.

Objective To understand whether different anesthesia and analgesia methods would affect the incidence of chronic postsurgical pain (CPSP) in patients who underwent uniportal video-assisted lung surgery and to explore the influence factors of CPSP. Design Retrospective study. Setting and Participant A total of 120 patients, who underwent selective uniportal video-assisted lung surgery at Zhongshan Hospital from March to June 2018, were enrolled. Interventions The visual analog scale was used to assess the degree of pain on the first and second postoperative days. At the 3rd and 6th months after surgery, the telephone follow-up was conducted for the survey of the numerical rating scale. The incidence of acute and chronic pain in different anesthesia methods was analyzed, and the relevant factors of CPSP were statistically analyzed. Results Among the 111 patients who completed follow-up, no significant difference was noted in the incidences of acute and chronic postsurgical pain between patients who received different anesthesia and analgesia methods (P>0.05). The incidence of CPSP was 29.7% at 3 months after surgery. The incidence of CPSP was 9.0% at 6 months after surgery. The degree of pain within 48 h after surgery was a risk factor for CPSP (odds ratio (OR)=2.39, P Conclusion CPSP accounted for a specific proportion of patients after uniportal video-assisted lung surgery; however, it was significantly lower than that of patients with conventional thoracotomy, and the incidence did not differ significantly among anesthesia and analgesia methods. Active and effective control of acute postsurgical pain might reduce the incidence of CPSP.

Hypoxia is a key factor in tumor microenvironments. Tumor-derived exosomes under hypoxia have their functions of communication between local and remote cells, and play an important role in tumor growth and metastasis. However, the effect of tumor-derived exosomes on cell structures and functions under hypoxia is unknown. In this work, the effects of exosomes derived from hepatocellular carcinoma cells (HCC-LM3) under normoxia (N-exos) and hypoxia (H-exos) environments on the biological and physical properties of target cells were stduied. The N-exos promoted the proliferation of hepatocytes (HL-7702) at 1.5 mg/mL, while the H-exos promoted the proliferation of hepatocytes at a lower concentration (1.0 mg/mL). After the cells cultured with the same concentrations of N-exos and H-exos for different time periods, the cell migration was enhanced. The stress fibers of the cells became loose and the cytoskeleton was rearranged, which were time dependent. The changes in morphological and mechanical parameters of the HL-7702 cells were detected by atomic force microscopy (AFM). The results showed that the cell edges became irregular and the filopodia were increased versus the exosomes treatment time. The heights and elastic moduli of cells were reduced. Compared with N-exos, H-exos had a more significant effect on the biological and physical properties of target cells. The results provide a method for studying how tumor-derived exosomes affect the interaction between tumor cells and their hypoxic microenvironment.

The objectives of this study were to improve the efficiency and accuracy of early clinical diagnosis of cervical cancer and to explore the application of tissue classification algorithm combined with multispectral imaging in screening of cervical cancer. 50 patients with suspected cervical cancer were selected. Firstly, the multispectral imaging technology was used to collect the multispectral images of the cervical tissues of 50 patients under the conventional white light waveband, the narrowband green light waveband, and the narrowband blue light waveband. Secondly, the collected multispectral images were fused, and then the tissue classification algorithm was used to segment the diseased area according to the difference between the cervical tissues without lesions and the cervical tissues with lesions. The difference in the contrast and other characteristics of the multiband spectrum fusion image would segment the diseased area, which was compared with the results of the disease examination. The average gradient, standard deviation (SD), and image entropy were adopted to evaluate the image quality, and the sensitivity and specificity were selected to evaluate the clinical application value of discussed method. The fused spectral image was compared with the image without lesions, it was found that there was a clear difference, and the fused multispectral image showed a contrast of 0.7549, which was also higher than that before fusion (0.4716), showing statistical difference ( P < 0.05 ). The average gradient, SD, and image entropy of the multispectral image assisted by the tissue classification algorithm were 2.0765, 65.2579, and 4.974, respectively, showing statistical difference ( P < 0.05 ). Compared with the three reported indicators, the values of the algorithm in this study were higher. The sensitivity and specificity of the multispectral image with the tissue classification algorithm were 85.3% and 70.8%, respectively, which were both greater than those of the image without the algorithm. It showed that the multispectral image assisted by tissue classification algorithm can effectively screen the cervical cancer and can quickly, efficiently, and safely segment the cervical tissue from the lesion area and the nonlesion area. The segmentation result was the same as that of the doctor's disease examination, indicating that it showed high clinical application value. This provided an effective reference for the clinical application of multispectral imaging technology assisted by tissue classification algorithm in the early screening and diagnosis of cervical cancer.

Circular RNAs (circRNAs) have been increasingly revealed to be desirable biomarkers for some tumors, including hepatocellular carcinoma (HCC). Combined with our previous microarray screening results, we aimed to determine the hsa_circ_0028502 and hsa_circ_0076251 expression features in HCC, analyze the relationship between their expression level and clinical and pathological characteristics, and investigate their diagnostic and prognostic values. Our data demonstrated that the hsa_circ_0028502 and hsa_circ_0076251 levels were considerably lower in HCC tissues than in adjacent paracancerous tissues (P, hsa_circ_0028502 and hsa_circ_0076251 are potentially valuable biomarkers for HCC diagnosis, and meanwhile, hsa_circ_0076251 could serve as a prognostic indicator for HCC.

Extracellular vesicles (EVs) secreted by cancer cells play a key role in the cancer microenvironment and progression. Previous studies have mainly focused on molecular functions, cellular components and biological processes using chemical and biological methods. However, whether the mechanical properties of cancer cells change due to EVs remains poorly understood. This work studies the effects of mechanical changes in non-small cell lung cancer (NSCLC) cells after treatment with EVs on migration by atomic force microscopy (AFM). Different concentrations of EVs were added into the experimental groups based on co-culture experiments, while the control group was cultured without EVs for 48 h. Cellular migration was evaluated by wound healing experiments. The cellular morphology, cell stiffness and surface adhesion were investigated by AFM. Cytoskeleton changes were detected by fluorescence staining assay. By comparison to the control group, the cell migration was enhanced. After treatment with EVs, the cell length and height show an upward trend, and the adhesion force and Young's modulus show a downward trend, and filopodia were also detected in the cells. Overall, the EVs promoted the migration of NSCLC cells by regulating cells' physical properties and skeletal rearrangement.

Tumor-derived exosomes (exos) are closely related to the occurrence, development and treatment of tumors. However, it is not clear how the exosomes affect the physical properties, which lead to the deterioration of the target cells. In this paper, atomic force microscopy (AFM) was used to study the effects of exosomes in HCC-LM3 cells and other cells (SMMC-7721 and HL-7702). The results showed that the HCC-LM3-exos (the exosomes secreted by HCC-LM3 cells, 50 μg mL-1) significantly promoted the proliferation and migration of HCC-LM3 cells. HCC-LM3-exos also promoted the proliferation and migration of SMMC-7721 and HL-7702 cells at 1000 and 1500 μg mL-1, respectively. With an increase in time and concentration, the proliferation effect was more significant. On comparing the mechanical properties of the three types of cells (HCC-LM3, SMMC-7721 and HL-7702 cells), the degradation degree and migration ability of the cells were from high to low in the above order. In turn, the surface roughness of the cells decreased, and adhesion and elastic modulus increased. With an increase in treatment time, surface roughness increased, while adhesion and elastic modulus decreased. These suggested that the HCC-LM3-exos could change the mechanical properties of cells, leading to their deterioration, and enhance their migration and invasion ability. In this paper, the effects of exosomes were analyzed from the perspective of the physical parameters of cells, which provide a new idea to study cancer metastasis and prognosis.

Chitinase 3-like protein 1(CHI3L1) participates in physiological and pathophysiological process, such as cell survival, cell proliferation, tissue remodeling, angiogenesis, etc. Some studies demonstrated that CHI3L1 is liver-enriched and has better application value in staging liver fibrosis than platelet ratio index(APRI) and fibrosis-4 index(FIB-4) and that CHI3L1 can be used in monitoring the prognosis of hepatocellular carcinoma (HCC). In this review, we summarized the pathophysiological role and the diagnostic value of CHI3L1 in liver fibrosis in different background and HCC.

Previous studies reported that exosomes (Exos) secreted by tumor cells could affect the tumor cells themselves and normal cells. However, the effects of exosomes derived from tumor cells on normal cells' migration and mechanical characteristics are rarely reported. This work explores the effects of H1299 cell-derived exosomes (H1299-Exos) on the migration of BEAS-2B cells, and analyzes possible mechanical mechanisms. In the experiments, exosomes were isolated from the culture supernatants of H1299 cells by ultracentrifugation. The H1299-Exos were confirmed by scanning electron microscope (SEM) and western blotting (WB). The BEAS-2B cell migration was assessed using scratch assays. Cytoskeletal structure changes were detected by immunofluorescence. Surface morphology and mechanical properties were measured by atomic force microscopy (AFM). After incubation with H1299-Exos for 48 h, BEAS-2B cells enhanced migration ability, with increased filopodia and cytoskeletal rearrangements. The changes in the morphology and mechanical properties of the cells caused by H1299-Exos were detected using AFM, including the increase in cell length and the decrease in cell height, Young's modulus and adhesion. In short, H1299-Exos promoted the BEAS-2B cell migrations. It indicates that the morphological and mechanical properties can be used as a means to assess normal cell alterations induced by tumor cell derived-exosomes. This provides a method for studying the effects of exosomes secreted by tumor cells on normal cells and the changes in their physical properties.

The urinary C-terminal telopeptide fragment of type II collagen (uCTX-II) has been reported as the efficient blood-based biomarker for osteoarthritis, which affects knees, hands, spine, and hips. This study reports a sensing strategy with antibody-conjugated gold nanoparticles (GNP) on an interdigitated electrode (IDE) to determine uCTX-II. The GNP-antibody complex was chemically immobilized on the IDE surface through the amine linker. uCTX-II was determined by monitoring the alteration in current upon interacting the GNP-complexed antibody. This strategy was improved the detection by attracting higher uCTX-II molecules, and the detection limit falls in the range of 10-100 pM with an acceptable regression value [y = 0.6254x - 0.4073, R² = 0.9787]. The sensitivity of the detection was recognized at 10 pM. Additionally, upon increasing the uCTX-II concentration, the current changes were increased in a linear fashion. Control detection with nonimmune antibody and control protein do not increase the current level, confirming the specific detection of uCTX-II. This method of detection helps in diagnosing osteoarthritis and its follow-up treatment.

Objective The aim of this study was to investigate the clinical characteristics of Corona Virus Disease 2019 in Taizhou, China. Methods A single center retrospective observational study was performed between Jan 1, 2020 and Mar 11, 2020 at Taizhou Public Health Medical Center, Zhejiang, China. All patients with confirmed Corona Virus Disease 2019 were enrolled, and their clinical data were gathered by reviewing electronic medical records. Outcomes of severely ill patients and non-severely ill patients were compared. Results Of 145 hospitalized patients with COVID-19, the average age was 47.5 years old (standard deviation, 14.6) and 54.5% were men. Hypertension was the most common comorbidity (15.2%), followed by diabetes mellitus (9.7%). Common symptoms included dry cough (81.4%), fever (75.2%), anorexia (42.8%), fatigue (40.7%), chest tightness (32.4%), diarrhea (26.9%) and dizziness (20%). According to imaging examination, 79.3% patients showed bilateral pneumonia, 18.6% showed unilateral pneumonia, 61.4% showed ground-glass opacity, and 2.1% showed no abnormal result. Compared with non-severely ill patients, severely ill patients were older (mean, years, 52.8 vs. 45.3, p p = 0.08), had a higher body mass index (mean, 24.78 vs. 23.20, p = 0.02) and were more likely to have fever (90.7% vs. 68.6%, p = 0.01), anorexia (60.5% vs. 35.3%, p = 0.01), chest tightness (60.5% vs.20.6%, p p = 0.03). Of the 43 severely ill patients, 6 (14%) received high-flow nasal cannula oxygen therapy, and 1 (2.3%) received invasive mechanical ventilation. Conclusions Older patients or patients with comorbidities such as obesity or diabetes mellitus were more likely to have severe condition. Treatments of COVID-19 is still experimental and more clinical trials are needed.

IntroductionTreatment failures for lupus nephritis (LN) are high with 10%–30% of patients progressing to end-stage renal disease (ESRD) within 10 years. Interstitial fibrosis/tubular atrophy (IFTA) is a predictor of progression to ESRD. Prior studies suggest that tubulointerstitial injury secondary to proteinuria in LN is mediated by complement activation in the tubules, specifically through the membrane attack complex (MAC). This study aimed to investigate the associations between tubular MAC deposition with IFTA and proteinuria.MethodsIn this cross-sectional study, LN kidney biopsies were assessed for MAC deposition by staining for Complement C9, a component of the MAC. Chromogenic immunohistochemistry was performed on paraffin-embedded human renal biopsy sections using unconjugated, murine anti-human Complement C9 (Hycult Biotech, clone X197). Tubular C9 staining intensity was analysed as present versus absent. IFTA was defined as minimal (50%).ResultsRenal biopsies from 30 patients with LN were studied. There were 24 (80%) female sex, mean age (SD) was 33 (12) years old and 23 (77%) had pure/mixed proliferative LN. Tubular C9 staining was present in 7 (23%) biopsies. 27 patients had minimal-to-mild IFTA and 3 patients had moderate IFTA. Among the C9 + patients, 3 (43%) had moderate IFTA as compared with none in the C9- group, p=0.009. C9 + patients had higher median (IQR) proteinuria as compared with C9- patients: 6.2 g (3.3–13.1) vs 2.4 g (1.3–4.6), p=0.001 at the time of biopsy. There was no difference in estimated glomerular filtration rate (eGFR) between the C9 + and C9- groups.ConclusionThis study demonstrated that tubular MAC deposition is associated with higher degree of IFTA and proteinuria, which are predictors of progression to ESRD. These results suggest that tubular MAC deposition may be useful in classification of LN. Understanding the role of complement in tubulointerstitial injury will also identify new avenues for LN treatment.

// Xiangyi He 1 , Jie Zhong 1 , Shuwei Wang 2 , Yufen Zhou 3 , Lei Wang 1 , Yongping Zhang 1 , Yaozong Yuan 1 1 Department of Gastroenterology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China 2 Department of Anesthesia, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China 3 Department of Gastroenterology, Ruijin Hospital North, Shanghai Jiaotong University School of Medicine, Shanghai, China Correspondence to: Yaozong Yuan, email: yyz28@medmail.com.cn Keywords: pancreatic cancer, metabolomics, serum, new-onset diabetes Received: May 31, 2016 Accepted: February 20, 2017 Published: March 16, 2017 ABSTRACT To establish a screening strategy for pancreatic cancer (PC) based on new-onset diabetic mellitus (NO-DM), serum metabolomics analysis and a search for the metabolic pathways associated with PC related DM were performed. Serum samples from patients with NO-DM (n = 30) and patients with pancreatic cancer and NO-DM were examined by liquid chromatography-mass spectrometry. Data were analyzed using principal components analysis (PCA) and orthogonal projection to latent structures (OPLS) of the most significant metabolites. The diagnostic model was constructed using logistic regression analysis. Metabolic pathways were analyzed using the web-based tool MetPA. PC patients with NO-DM were older and had a lower BMI and shorter duration of DM than those with NO-DM. The metabolomic profiles of patients with PC and NO-DM were significantly different from those of patients with NO-DM in the PCA and OPLS models. Sixty two differential metabolites were identified by the OPLS model. The logistic regression model using a panel of two metabolites including N_Succinyl_L_diaminopimelic_acid and PE (18:2) had high sensitivity (93.3%) and specificity (93.1%) for PC. The top three metabolic pathways associated with PC related DM were valine, leucine and isoleucine biosynthesis and degradation, primary bile acid biosynthesis, and sphingolipid metabolism. In conclusion, screening for PC based on NO-DM using serum metabolomics in combination with clinic characteristics and CA19-9 is a potential useful strategy. Several metabolic pathways differed between PC related DM and type 2 DM.

Background. Nifedipine-induced gingival overgrowth (NGO) is a multifactorial pathogenesis with increased extracellular matrix including collagen and glycans, inflammatory cytokines, and phenotype changes of fibroblasts. However, the molecular etiology of NGO is not well understood. The objective of this study is to investigate the key genes in the pathogenesis of NGO. Methods. In this study, we examined the proliferation and migration abilities of fibroblasts derived from patients with chronic periodontitis, nifedipine nonresponder gingival overgrowth, gingival overgrowth caused by nifedipine, and healthy normal gingiva. We conducted RNA-Seq on these four groups of fibroblasts and analysed the differentially expressed genes (DEGs). Results. Fibroblasts derived from NGO patients had higher proliferation and migration abilities than those of the other groups. Protein-protein interaction network analysis indicated that TGFB2, ITGA8, ITGA11, FGF5, PLA2G4D, PLA2G2F, PTGS1, CSF1, LPAR1, CCL3, and NKX3-1 are involved in the development of NGO. These factors are related to the arachidonic acid metabolism and PI3K/AKT signaling pathways. Conclusion. Transcriptional gene expression analysis identified a number of DEGs that might be functionally related to gingival overgrowth induced by nifedipine. Our study provides important information on the molecular mechanism underlying nifedipine-induced gingival overgrowth.

Aim In the present study, we purposed to determine serum chitinase 3‐like 1 (CHI3L1) expression characteristics in chronic liver diseases monoinfected with hepatitis B virus and analyze its diagnostic value in liver fibrosis. Methods A total of 467 chronic hepatitis B (CHB) patients, 312 liver cirrhosis (LC) patients, and 104 hepatocellular carcinoma (HCC) patients at our institution were enrolled, and clinical indicators were analyzed. Results Our data have shown that the expression level of serum CHI3L1 was steadily increased from CHB to LC to HCC (P

// Yuanfen Zhai 1,* , Rongbin Wei 1,* , Junjun Liu 1 , Huihui Wang 2 , Wenping Cai 1 , Mengmeng Zhao 1 , Yongguang Hu 1 , Shuwei Wang 3 , Tianshu Yang 1 , Xiaodong Liu 4 , Jianhua Yang 1 and Shangfeng Liu 1,3 1 Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, P. R. China 2 Department of Pediatric Dentistry, School of Stomatology, Tongji University, Shanghai Engineering Research Center, Shanghai, P. R. China 3 Department of Stomatology, Huashan Hospital, Fudan University, Shanghai, P. R. China 4 Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, P. R. China * These authors have contributed equally to this work Correspondence to: Shangfeng Liu, email: // Jianhua Yang, email: // Xiaodong Liu, email: // Keywords : aging; dental stem cells; cellular senescence model; stress; DNA damage; Gerotarget Received : September 18, 2016 Accepted : December 12, 2016 Published : December 21, 2016 Abstract Aging is identified by a progressive decline of physiological integrity leading to age-related degenerative diseases, but its causes is unclear. Human dental pulp stem cells (hDPSCs) has a remarkable rejuvenated capacity that relies on its resident stem cells. However, because of the lack of proper senescence models, exploration of the underlying molecular mechanisms has been hindered. Here, we established a cellular model utilizing a hydroxyurea (HU) treatment protocol and effectively induced Human dental pulp stem cells to undergo cellular senescence. Age-related phenotypic changes were identified by augmented senescence-associated-β-galactosidase (SA-β-gal) staining, declined proliferation and differentiation capacity, elevated G0/G1 cell cycle arrest, increased apoptosis and reactive oxygen species levels. Furthermore, we tested the expression of key genes in various DNA repair pathways including nonhomologous end-joining (NHEJ) and homologous recombination (HR) pathways. In addition, our results showed that Dental pulp stem cells from young donors are more resistant to apoptosis and exhibit increased non-homologous end joining activity compared to old donors. Further transcriptome analysis demonstrate that multiple pathways are involved in the HU-induced Dental pulp stem cells ageing, including genes associated with DNA damage and repair, mitochondrial dysfunction and increased reactive oxygen species levels. Taken together, the cellular model have important implications for understanding the molecular exploration of Dental pulp stem cells senescence and aging.

Circular RNAs (circRNAs) are formed from the genome through diverse back splicing and feature the closed loop. circRNAs are widely available in a variety of cells and characterized by conservation, structural stability, high abundance and tissue-specific or developmental-specific expression. Recent studies have shown that circRNAs are closely related to liver diseases, such as metabolic-associated fatty liver disease, hepatitis, liver cirrhosis and hepatocellular carcinoma. circRNAs play an important role in the progression of liver diseases, are potential diagnostic and prognostic markers, and have translational value in therapy. This article reviews the research on circRNAs in liver diseases, with a view to providing a theoretical basis and new ideas for future research and treatment of liver diseases.

Glioma is the most common and aggressive type of brain tumor. It has a poor prognosis and a high recurrence rate. Despite continued advances in surgery, chemotherapy and radiotherapy, the clinical outcomes remain dismal. MicroRNAs (miRNAs) play important roles in the initiation and progression of a multitude of tumors. Until now, the molecular mechanism that is responsible for glioma tumorigenesis and progression remains unclear. Increasing evidence has shown that miRNAs play an important role in glioma. In this review, we focus on the current advances in determining the role of miRNAs in regulating tumorigenesis and the progression of glioma. In addition, the relevant roles of miRNAs about the diagnosis and target therapy have been clarified.

Neuronal K(v)7/KCNQ channels are voltage-gated potassium channels composed of K(v)7.2/KCNQ2 and K(v)7.3/KCNQ3 subunits. Enriched at the axonal membrane, they potently suppress neuronal excitability. De novo and inherited dominant mutations in K(v)7.2 cause early onset epileptic encephalopathy characterized by drug resistant seizures and profound psychomotor delay. However, their precise pathogenic mechanisms remain elusive. Here, we investigated selected epileptic encephalopathy causing mutations in calmodulin (CaM)-binding helices A and B of K(v)7.2. We discovered that R333W, K526N, and R532W mutations located peripheral to CaM contact sites decreased axonal surface expression of heteromeric channels although only R333W mutation reduced CaM binding to K(v)7.2. These mutations also altered gating modulation by phosphatidylinositol 4,5-bisphosphate (PIP(2)), revealing novel PIP(2) binding residues. While these mutations disrupted K(v)7 function to suppress excitability, hyperexcitability was observed in neurons expressing K(v)7.2-R532W that displayed severe impairment in voltage-dependent activation. The M518V mutation at the CaM contact site in helix B caused most defects in K(v)7 channels by severely reducing their CaM binding, K(+) currents, and axonal surface expression. Interestingly, the M518V mutation induced ubiquitination and accelerated proteasome-dependent degradation of K(v)7.2, whereas the presence of K(v)7.3 blocked this degradation. Furthermore, expression of K(v)7.2-M518V increased neuronal death. Together, our results demonstrate that epileptic encephalopathy mutations in helices A and B of K(v)7.2 cause abnormal K(v)7 expression and function by disrupting K(v)7.2 binding to CaM and/or modulation by PIP(2). We propose that such multiple K(v)7 channel defects could exert more severe impacts on neuronal excitability and health, and thus serve as pathogenic mechanisms underlying Kcnq2 epileptic encephalopathy.

Objective To investigate the clinical significance of long non-coding RNA LINC01617 in esophageal cancer and explore the effect of LINC01617 on the proliferation and metastasis of esophageal cancer cells. Methods Real time fluorescence PCR was used to detect the expression of LINC01617 in 142 cases of esophageal cancer and adjacent tissues. The relationship between the expression level of LINC01617 and the survival rate of esophageal cancer patients was analyzed. The function of LINC01617 was detected in esophageal cancer cell lines. The tumor growth ability test was carried out in the nude mice. Results We found that LINC01617 was overexpressed in esophageal cancer, and its expression was associated with poor prognosis of esophageal cancer. In vitro experiments confirmed that knockout of LINC01617 significantly inhibited the proliferation, migration and invasion of esophageal cancer cells. Moreover, knockout of LNC01617 can inhibit the growth of esophageal cancer in nude mice. The Akt pathway may be involved in the regulation of cell activity in esophageal cancer. Conclusions LINC01617 may be involved in the occurrence and development of esophageal cancer, suggesting that LINC01617 can be used as a biomarker and potential therapeutic target for esophageal cancer.

SummaryContext Pituitary stalk interruption syndrome (PSIS) is a rare cause of combined pituitary hormone deficiency characterized by a triad shown in pituitary imaging, yet it has never been evaluated due to the visibility of pituitary stalk (PS) in imaging findings. Objective The major objective of the study was to systematically describe the disease including clinical presentations, imaging findings and to estimate the severity of anterior pituitary hormone deficiency based on the visibility of the PS. Methods This was a retrospective study including 74 adult patients with PSIS in Shanghai Clinical Center for Endocrine and Metabolic Diseases between January 2010 and June 2014. Sixty had invisible PS according to the findings on MRI, while the rest had a thin or intersected PS. Basic characteristics and hormonal status were compared. Results Of the 74 patients with PSIS, age at diagnosis was 25 (22–28) years. Absent pubertal development (97·3%) was the most common presenting symptom, followed by short stature. Insulin tolerance test (ITT) and gonadotrophin-releasing hormone (GnRH) stimulation test were used to evaluate the function of anterior pituitary. The prevalence of isolated deficiency in growth hormone (GH), gonadotrophins, corticotrophin and thyrotrophin were 100%, 97·2%, 88·2% and 70·3%, respectively. Although the ratio of each deficiency did not vary between patients with invisible PS and with visible PS, panhypopituitarism occurred significantly more frequent in patients with invisible PS. Patients with invisible PS had significantly lower levels of luteinizing hormone (LH), follicle stimulation hormone (FSH) and hormones from targeted glands including morning cortisol, 24-h urine free cortisol, free triiodothyronine (FT3), free thyroxine (FT4) and testosterone (T) in male than patients with visible PS. Moreover, patients with invisible PS had lower peak LH and FSH in GnRH stimulation test, and higher peak cortisol in ITT while peak GH remained unchanged between two groups. Conclusions The prevalence of multiple anterior pituitary hormone deficiency was high in adult patients with PSIS. And more importantly, we found the visibility of PS shown on MRI might be an indication of the severity of PSIS.

// Nan Wang 1 , Dan Guo 1 , YangYang Zhao 1 , ChengYa Dong 1 , XiaoYan Liu 1 , BinXia Yang 1 , ShuWei Wang 1 , Lin Wang 1 , QingGuo Liu 1 , Qian Ren 1 , YongMin Lin 1 , XiaoTong Ma 1 1 State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, China Correspondence to: XiaoTong Ma, e-mail: maxt@ihcams.ac.cn Keywords: TWIST-1, myeloid leukemia, leukemia stem cell, prognostic factor, c-MPL Received: December 11, 2014 Accepted: May 08, 2015 Published: May 20, 2015 ABSTRACT Alterations of TWIST-1 expression are often seen in solid tumors and contribute to tumorigenesis and cancer progression. However, studies concerning its pathogenic role in leukemia are scarce. Our study shows that TWIST-1 is overexpressed in bone marrow mononuclear cells of patients with acute myeloid leukemia (AML) and chronic myeloid leukemia (CML). Gain-of-function and loss-of-function analyses demonstrate that TWIST-1 promotes cell growth, colony formation and drug resistance of AML and CML cell lines. Furthermore, TWIST-1 is aberrantly highly expressed in CD34 + CD38 − leukemia stem cell candidates and its expression declines with differentiation. Down-modulation of TWIST-1 in myeloid leukemia CD34 + cells impairs their colony-forming capacity. Mechanistically, c-MPL, which is highly expressed in myeloid leukemia cells and associated with poor prognosis, is identified as a TWIST-1 coexpressed gene in myeloid leukemia patients and partially contributes to TWIST-1-mediated leukemogenic effects. Moreover, patients with higher TWIST-1 expression have shorter overall and event-free survival (OS and EFS) in AML. Multivariate analysis further demonstrates that TWIST-1 overexpression is a novel independent unfavourable predictor for both OS and EFS in AML. These data highlight TWIST-1 as a new candidate gene contributing to leukemogenesis of myeloid leukemia, and propose possible new avenues for improving risk and treatment stratification in AML.

Background/aim MicroRNA-93-5p (miR-93-5p) dysregulation has been reported in many types of human cancer. However, the collective effect of miR-93-5p in both lung adenocarcinoma and squamous cell carcinoma and the mechanism underlying miR-93-5p involvement in non-small cell lung cancer cells (NSCLC) is unknown. Herein, our purpose was to reveal the role and explain this mechanism, with the goal of contributing to the development of new diagnostic biomarkers and individualized therapeutic targets. Materials and methods We examined miR-93-5p expression in NSCLC specimens (including lung adenocarcinoma and squamous cell carcinoma) by qPCR. The effects of miR-93-5p inhibitor on proliferation, migration, and invasion of NSCLC cells were determined by MTT assay, colony formation assays, apoptosis assay, cell cycle assay and transwell chamber assays, respectively. The molecular mechanisms underlying these effects were investigated via dual luciferase reporter assay and western blotting. Results MiR-93-5p expression levels were significantly correlated with NSCLC patients overall survival rate. Cell proliferation, migration, and invasion were significantly inhibited by miR-93-5p down-regulation. Dual luciferase reporter assay demonstrated that miR-93-5p directly bound with the 3′-untranslated region of the tumor suppressor gene PTEN and RB1. Conclusion MiR-93-5p is up-regulated in NSCLC and plays an oncogenic role by inhibiting PTEN and RB1, suggesting miR-93-5p may be a novel prognostic indicator and a therapeutic target in NSCLC.

Fibroblast growth factor receptors are growth factor receptor tyrosine kinases, exerting their roles in embryogenesis, tissue homeostasis, and development of breast cancer. Recent genetic studies have identified some subtypes of fibroblast growth factor receptors as strong genetic loci associated with breast cancer. In this article, we review the recent epidemiological findings and experiment results of fibroblast growth factor receptors in breast cancer. First, we summarized the structure and physiological function of fibroblast growth factor receptors in humans. Then, we discussed the common genetic variations in fibroblast growth factor receptors that affect breast cancer risk. In addition, we also introduced the potential roles of each fibroblast growth factor receptors isoform in breast cancer. Finally, we explored the potential therapeutics targeting fibroblast growth factor receptors for breast cancer. Based on the biological mechanisms of fibroblast growth factor receptors leading to the pathogenesis in breast cancer, targeting fibroblast growth factor receptors may provide new opportunities for breast cancer therapeutic strategies.

Background The effectiveness of psoriasis therapies in real-world settings remains relatively unknown. Objective We sought to compare the effectiveness of less commonly used systemic therapies and commonly used combination therapies for psoriasis. Methods This was a multicenter cross-sectional study of 203 patients with plaque psoriasis receiving less common systemic monotherapy (acitretin, cyclosporine, or infliximab) or common combination therapies (adalimumab, etanercept, or infliximab and methotrexate) compared with 168 patients receiving methotrexate evaluated at 1 of 10 US outpatient dermatology sites participating in the Dermatology Clinical Effectiveness Research Network. Results In adjusted analyses, patients on acitretin (relative response rate 2.01; 95% confidence interval [CI] 1.18-3.41), infliximab (relative response rate 1.93; 95% CI 1.26-2.98), adalimumab and methotrexate (relative response rate 3.04; 95% CI 2.12-4.36), etanercept and methotrexate (relative response rate 2.22; 95% CI 1.25-3.94), and infliximab and methotrexate (relative response rate 1.72; 95% CI 1.10-2.70) were more likely to have clear or almost clear skin compared with patients on methotrexate. There were no differences among treatments when response rate was defined by health-related quality of life. Limitations Single time point assessment may result in overestimation of effectiveness. Conclusions The efficacy of therapies in clinical trials may overestimate their effectiveness as used in clinical practice. Although physician-reported relative response rates were different among therapies, absolute differences were small and did not correspond to differences in patient-reported outcomes.

Cancer cells are characterized by altered glucose metabolism known as the Warburg effect in which aerobic glycolysis is increased. Glucose is converted to lactate even under sufficient oxygen tension. Interfering with this process may be a potential effective strategy to cause cancer cell death because these cells rely heavily on glucose metabolism for survival and proliferation. 2-Deoxy-D-glucose (2DG), a glucose analog, targets glucose metabolism to deplete cancer cells of energy. In addition, 2DG increases oxidative stress, inhibits N-linked glycosylation, and induces autophagy. It can efficiently slow cell growth and potently facilitate apoptosis in specific cancer cells. Although 2DG itself has limited therapeutic effect in many types of cancers, it may be combined with other therapeutic agents or radiotherapy to exhibit a synergistic anticancer effect. In this review, we describe the Warburg effect and discuss 2DG and its underlying mechanisms and potential application for cancer treatment.

Interferon‑induced transmembrane protein 3 (IFITM3) has been recently identified as a potential molecular marker. IFITM3 has been reported to be upregulated in various human diseases, including colon and breast cancer, astrocytoma, as well as ulcerative colitis. However, the clinical significance and underlying mechanisms of IFITM3 dysregulated expression in gastric cancer (GC) remain to be determined. The present study aimed to evaluate the expression of IFITM3 in human gastric tumor cells and specimens and investigate the effects of IFITM3 knockdown in the regulation of GC growth and its potential mechanism. IFITM3 expression was significantly overexpressed in the GC cell lines and GC tissues compared with corresponding normal controls by RT‑qPCR, western blot analysis and immunohistochemistry, and this overexpression was correlated with tumor differentiation, lymph node and distant metastasis, and advanced tumor node metastasis stages. Furthermore, knockdown of IFITM3 expression suppressed tumor cell migration, invasion and proliferation significantly in vitro, arrested tumor cells at the G0/G1 phase and reduced the cell numbers in the S phase of the cell cycle. We preliminarily confirmed that IFITM3 can be mediated by the activities of Wnt/β-catenin signaling. Further investigation revealed that silencing of IFITM3 effectively reversed the epithelial-to-mesenchymal transition (EMT) phenotype and reduced the activities of MMP-2 and MMP-9 expression. Taken together, these data suggested that IFITM3 is a potential therapeutic target for GC.

Interleukin (IL)-6 and the downstream Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway have previously been reported to be important in the development of colorectal cancer (CRC), and several studies have shown the relationship between the polymorphisms of related genes in this pathway with the risk of CRC. However, the findings of these related studies are inconsistent. Moreover, there has no systematic review and meta-analysis to evaluate the relationship between genetic variants in IL-6/JAK/STAT3 pathway and CRC susceptibility. Hence, we conducted a meta-analysis to explore the relationship between polymorphisms in IL-6/JAK/STAT3 pathway genes and CRC risk. Eighteen eligible studies with a total of 13,795 CRC cases and 18,043 controls were identified by searching PubMed, Web of Science, Embase, and the Cochrane Library databases for the period up to September 15, 2015. Odds ratios (ORs) and their 95 % confidence intervals (CIs) were used to calculate the strength of the association. Our results indicated that IL-6 genetic variants in allele additive model (OR = 1.05, 95 % CI = 1.00, 1.09) and JAK2 genetic variants (OR = 1.40, 95 % CI = 1.15, 1.65) in genotype recessive model were significantly associated with CRC risk. Moreover, the pooled data revealed that IL-6 rs1800795 polymorphism significantly increased the risk of CRC in allele additive model in Europe (OR = 1.07, 95 % CI = 1.01, 1.14). In conclusion, the present findings indicate that IL-6 and JAK2 genetic variants are associated with the increased risk of CRC while STAT3 genetic variants not. We need more well-designed clinical studies covering more countries and population to definitively establish the association between genetic variants in IL-6/JAK/STAT3 pathway and CRC susceptibility.

Importance Hypertension is prevalent among patients with psoriasis. The effect of psoriasis and its severity on hypertension control is unknown. Objective To determine the association between uncontrolled blood pressure and psoriasis, both overall and according to objectively measured psoriasis severity, among patients with diagnosed hypertension. Design, Setting, and Participants Population-based cross-sectional study nested in a prospective cohort drawn from The Health Improvement Network (THIN), an electronic medical records database broadly representative of the general population in the United Kingdom. The study population included a random sample of patients with psoriasis (n = 1322) between the ages of 25 and 64 years in THIN who were included in the Incident Health Outcomes and Psoriasis Events prospective cohort and their age- and practice-matched controls without psoriasis (n = 11 977). All included patients had a diagnosis of hypertension; their psoriasis diagnosis was confirmed and disease severity was classified by their general practitioners. Main Outcomes and Measures Uncontrolled hypertension was defined as a systolic blood pressure of 140 mm Hg or higher or a diastolic blood pressure of 90 mm Hg or higher based on the blood pressure recorded closest in time to the assessment of psoriasis severity. Results There was a significant positive dose-response relationship between uncontrolled hypertension and psoriasis severity as objectively determined by the affected body surface area in both unadjusted and adjusted analyses that controlled for age, sex, body mass index, smoking and alcohol use status, presence of comorbid conditions, and current use of antihypertensive medications and nonsteroidal anti-inflammatory drugs (adjusted odds ratio [aOR], 0.97; 95% CI, 0.82-1.14 for mild psoriasis; aOR, 1.20; 95% CI, 0.99-1.45 for moderate psoriasis; and aOR, 1.48; 95% CI, 1.08-2.04 for severe psoriasis; P = .01 for trend). The likelihood of uncontrolled hypertension among psoriasis overall was also increased, although not statistically significantly so (aOR, 1.10; 95% CI, 0.98-1.24). Conclusions and Relevance Among patients with hypertension, psoriasis was associated with a greater likelihood of uncontrolled hypertension in a dose-dependent manner, with the greatest likelihood observed among those with moderate to severe psoriasis defined by 3% or more of the body surface area affected. Our data suggest a need for more effective blood pressure management, particularly among patients with more severe psoriasis.

To date, many studies have shown that microRNAs (miRNA) exhibit altered expression levels in various cancers and may play a potential role as diagnostic and prognostic biomarkers of cancers. This meta-analysis was designed to evaluate the exact role of microRNA-31 (miR-31) for survival and discuss the possibility of utilizing miR-31 to predict the prognosis of patients with various human cancers. Electronic literature databases including PubMed, Web of Science, and Embase were searched for articles published until May 2014. The articles only written in English were considered. Data were extracted from studies comparing overall survival (OS), cancer-specific survival (CSS), or postoperative survival (PS) in patients with multiple cancers, which showed higher miR-31 expression than with similar patients. Pooled hazard ratios (HRs) of miR-31 for survival and 95 % confidence intervals (CI) were calculated. Ten studies with a total of 1,648 participants were included for the meta-analysis. For OS, the pooled HRs of higher miR-31 expression in cancers indicated significant predictor poorer survival in general cancers in either univariate analysis (HR=2.34, 95 % CI=1.15-3.52, P

Interleukin (IL)-6 and the downstream Janus kinase (JAK)/signal activator of transcription (STAT) pathway have been found to be important in the development of colorectal cancer (CRC). To develop novel therapies for CRC, we have explored the effects of a novel small-molecule JAK inhibitor (AZD1480) on IL-6/JAK/STAT3 pathway and its potential anti- tumor activity on the human CRC cell lines (HCT116, HT29 and SW480). The results showed that, AZD1480 effectively prevents constitutive and IL-6-induced JAK2 and STAT-3 phosphorylation and exerted antitumor functional effects by a decrease in proliferation and an increase in apoptosis in CRC cells. The inhibition of tumorigenesis was consistent with the decreased phosphorylated JAK2 and phosphorylated STAT3, and the decreased expression of STAT3-targeted genes c-Myc, cyclin D2 and IL-6. Thus, AZD1480 is a potential new clinical therapeutic agent for patients with CRC.

Objective To determine the risk of chronic kidney disease in patients with psoriasis. Design Population based cohort study and nested cross sectional study. Setting Electronic medical records database based in United Kingdom. Participants Cohort study: patients with psoriasis aged 18-90 each matched to up to five patients without psoriasis based on age, practice, and time of visit. Nested study: patients with psoriasis aged 25-64 with confirmed data on psoriasis severity, each matched to up to 10 patients without psoriasis based on age and practice. Main outcome measures Cohort study: incident moderate to advanced (stage 3 through 5) chronic kidney disease. Nested study: baseline prevalence of chronic kidney disease. Results 136 529 patients with mild psoriasis and 7354 patients with severe psoriasis based on treatment patterns were matched to 689 702 unaffected patients. The adjusted hazard ratios (95% confidence intervals) for incident chronic kidney disease were 1.05 (1.02 to 1.07), 0.99 (0.97 to 1.02), and 1.93 (1.79 to 2.08) in the overall, mild, and severe psoriasis groups, respectively. Age was a significant effect modifier in the severe psoriasis group, with age specific adjusted hazard ratios (95% confidence intervals) of 3.82 (3.15 to 4.64) and 2.00 (1.86 to 2.17) for patients aged 30 and 60, respectively. In the nested analysis of 8731 patients with psoriasis with measurements of affected body surface area matched to 87 310 patients without psoriasis, the adjusted odds ratios (95% confidence intervals) for chronic kidney disease were 0.89 (0.72 to 1.10), 1.36 (1.06 to 1.74), and 1.58 (1.07 to 2.34) in the mild, moderate, and severe psoriasis groups, respectively. Conclusions Moderate to severe psoriasis is associated with an increased risk of chronic kidney disease independent of traditional risk factors.

Long non-coding RNAs (lncRNAs) are emerging as key molecules in human cancer. Highly upregulated in liver cancer (HULC), an lncRNA, has recently been revealed to be involved in hepatocellular carcinoma development and progression. It remains unclear, however, whether HULC plays an oncogenic role in human gastric cancer (GC). In the present study, we demonstrated that HULC was significantly overexpressed in GC cell lines and GC tissues compared with normal controls, and this overexpression was correlated with lymph node metastasis, distant metastasis and advanced tumor node metastasis stages. In addition, a receiver operating characteristic (ROC) curve was constructed to evaluate the diagnostic values and the area under the ROC curve of HULC was up to 0.769. To uncover its functional importance, gain- and loss-of-function studies were performed to evaluate the effect of HULC on cell proliferation, apoptosis and invasion in vitro. Overexpression of HULC promoted proliferation and invasion and inhibited cell apoptosis in SGC7901 cells, while knockdown of HULC in SGC7901 cells showed the opposite effect. Mechanistically, we discovered that overexpression of HULC could induce patterns of autophagy in SGC7901 cells; more importantly, autophagy inhibition increased overexpression of HULC cell apoptosis. We also determined that silencing of HULC effectively reversed the epithelial-to-mesenchymal transition (EMT) phenotype. In summary, our results suggest that HULC may play an important role in the growth and tumorigenesis of human GC, which provides us with a new biomarker in GC and perhaps a potential target for GC prevention, diagnosis and therapeutic treatment.

We thank Mr. Davis and Dr. Feldman for their insightful comments in response to our study. They addressed how the Hawthorne effect, which occurs when patients' behaviors change (such as improving adherence) after knowing they are being watched in ways that may improve outcomes, could be leveraged in clinical practice. In our cross-sectional study, patients were evaluated at a single time point under real world conditions (e.g. routine follow-up visit) and did not know that the effectiveness of their psoriasis treatment was going to be formally assessed until the day of their regularly scheduled clinic visit 1. Thus the Hawthorne effect could not have affected adherence and was unlikely to have impacted our estimates of physician reported outcomes but may have influenced patient reported outcomes. The degree to which suboptimal adherence with systemic psoriasis treatments influences effectiveness remains largely unknown. Nevertheless, optimizing patient adherence remains an important treatment goal and thus dermatologists may consider attempts at replicating the Hawthorne Effect in their clinical practices. However, achieving the Hawthorne effect in clinical practice may not be feasible for many practitioners and patients. More frequent follow-up office visits may be impractical due to various factors including time and expense for the patient and scheduling difficulties for the physician. Promising novel approaches to mimicking the type of close follow-up which occurs in the clinical trial setting include mobile tele-dermatology for home monitoring of psoriasis patients, which has been shown to have high acceptance ratings in both patients and dermatologists, with great potential for increasing patient motivation and compliance 2. We also note that the factors which explain differences in efficacy observed in clinical trials and effectiveness observed in real world settings are complex and multi-factorial. For example, patients on systemic psoriasis treatments often face issues of access (treatment expense, delays in obtaining insurance approvals to continue treatment or lack of insurance approval to increase the dose of treatment due to sub-optimal response, difficulty in traveling to offices that provide psoriasis treatment, etc.), tachyphylaxis, and often patients express safety concerns that result in treatment discontinuation 3. Additionally, prior treatment experience, which may inform response rates may not be reflected in clinical trials (i.e CHAMPION and ACCEPT trials included only TNF inhibitor naive patients whereas nearly 30% of our patients had used a different TNF inhibitor prior to their current TNF treatment) 4, 5. The determinants of psoriasis treatment effectiveness in real world settings remain an important scientific knowledge gap. Prospective effectiveness studies are urgently needed in order to optimize the quality of care for patients with psoriasis.

The aim of this study was to observe the protein changes of intestinal epithelial cells induced in vitro by Trichinella spiralis infective larvae and their excretory-secretory (ES) or surface antigens and identity the proteins related with invasion. HCT-8 cells were incubated for 2 h in the culture medium contained ES or surface antigens of infective larvae, and observed by Immunofluorescent test (IFT). The infective larvae were inoculated into culture of HCT-8 cells to incubate for 18 h, and the lysates of HCT-8 cells were analyzed by SDS-PAGE and Western blot. IFA showed that normal HCT-8 cells had positively reactions with sera of the infected mice and mice immunized with ES or surface antigens. However, after incubating with ES or surface antigens, HCT-8 cells had stronger positively reaction with the above sera. On Western blot, after cultured with infective larvae, additional seven protein bands (66, 61, 57, 45, 34, 21, and 17 kDa) of HCT-8 cells were recognized by sera of the infected or immunized mice, but three protein bands (48, 43, and 23 kDa) of HCT-8 cells were not recognized by the above sera, compared with normal HCT-8 cells. Our results showed that after cultured with infective larvae the protein components of HCT-8 cell changed, suggesting that additional seven proteins recognized by sera of the infected or immunized mice may be related with invasion of intestinal epithelial cells by infective larvae, these proteins might mediate or facilitate entry into the cells, while the three proteins not recognized by the above sera may be the specific mediators released from the cells which permit invasion.

ADAMTS13 limits platelet-rich thrombosis by cleaving von Willebrand factor at the Tyr(1605)-Met(1606) bond. Previous studies showed that ADAMTS13 truncated after spacer domain remains proteolytically active or hyperactive. However, the relative contribution of each domain within the proximal carboxyl terminus of ADAMTS13 in substrate recognition and specificity is not known. We showed that a metalloprotease domain alone was unable to cleave the Tyr-Met bond of glutathione S-transferase (GST)-VWF73-H substrate in 3 h, but it did cleave the substrate at a site other than the Tyr-Met bond after 16-24 h of incubation. Remarkably, the addition of even one or several proximal carboxyl-terminal domains of ADAMTS13 restored substrate specificity. Full proteolytic activity, however, was not achieved until all of the proximal carboxyl-terminal domains were added. The addition of TSP1 2-8 repeats and two CUB domains did not further increase proteolytic activity. Furthermore, ADAMTS13 truncated after the spacer domain with or without metalloprotease domain bound GST-VWF73-H with a K(d) of approximately 7.0 or 13 nm, comparable with full-length ADAMTS13 (K(d) = 4.6 nm). Metalloprotease domain did not bind GST-VWF73-H detectably, but the disintegrin domain, first TSP1 repeat, Cys-rich domain, and spacer domain bound GST-VWF73-H with K(d) values of 489, 136, 121, and 108 nm, respectively. These proximal carboxyl-terminal domains dose-dependently inhibited cleavage of fluorescent resonance energy transfer (FRETS)-VWF73 by full-length ADAMTS13 and ADAMTS13 truncated after the spacer domain. These data demonstrated that the proximal carboxyl-terminal domains of ADAMTS13 determine substrate specificity and are all required for recognition and cleavage of von Willebrand factor between amino acid residues Asp(1595) and Arg(1668).