OBJECTIVEThe aim of this study was to investigate the expression of connexin (Cx) and the function of gap junction intercellular communication (GJIC) in the carcinogenesis, progression and metastasis of gastric cancers.

METHODSImmunohistochemistry was used to detect the expression of Cx32 and Cx43 proteins in tissue samples. Indirect immunofluorescence assay was used to investigate the expression of Cx32 and Cx43 proteins in several gastric cancer lines of various differentiation grades. The expression of Cx43 in samples of gastric cancer tissue, adjacent normal tissue and in the gastric cancer cell lines of various differentiation grades was detected by Western blot. Scrape-loading dye transfer (SLDT) technique was used to detect the function of gap junction intercellular communication (GJIC) in the various cell lines.

RESULTSIn the normal gastric mucosa the expression rates of both Cx32 and Cx43 were 100%. In gastric cancers, the expression rates of Cx32 andCx43 were 49.5% (55/111) and 39.6% (44/111), respectively. There was a significant difference between their expression in normal and cancer tissues (P < 0.05). Age of the patients was not significantly correlated with the expression level of Cx32 and Cx43 (P > 0.05). Cx43 expression was significantly associated with the TMN stage, histological type, depth of infiltration and distant metastasis (P < 0.05), but Cx32 expression was not significantly correlated with depth of infiltration ( P > 0.05). In the cancer cell lines, a positive expression of Cx32 and Cx43 was detected in transfected human stomach mucosal cell line (CES-1) and human well differentiated stomach cancer cell line (N87), but negative in the poorly differentiated stomach cancer cell line (BGC-823) at all. Both Cx32 and Cx43 expression rates were 100% in the cell line GES-1. Cx32 expression rate was 49.0% and Cx43 expression rate was 55.0% in the cell line N87. But in the poorly differentiated cancer cell line BGC-823 both Cx32 and Cx43 expression was negative. GJIC function detection showed: GES-1 showed well GIJC function but no GIJC function in the cell lines N87 and BGC-823. The intensity of fluorescence was gradually decreasing from GES-1 cells to N87 cells and almost no fluorescence in BGC-823 cells. Western blotting showed that Cx43 expression in normal tissue was higher than that in gastric cancer tissue, and in the cell lines GES-1, N87 and BGC-823, the bands seemed decreasing progressively. There was very low expression in BGC-823 cells.

CONCLUSIONThe decreasing expression of connexin Cx32 and Cx43 is obviously correlated with the occurrence, development and metastatic potential of stomach cancers.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Cell Communication ; physiology ; Cell Line, Tumor ; Connexin 43 ; metabolism ; Connexins ; metabolism ; Down-Regulation ; Female ; Gap Junctions ; physiology ; Gastric Mucosa ; metabolism ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo explore the effect of changes of processing time on the surface properties of titanium coating formed by micro-arc oxidation (MAO).

METHODSForty-four disc-shaped pure titanium specimens with 10 mm diameter and 1 mm thickness were equally divided into 4 groups and processed by MAO technique in electrolytes containing 0.2 mol/L calcium acetate (CA) and 0.02 mol/L β-glycerol phosphate disodium salt pentahydrate (β-GP). The processing time were set at 1 min, 5 min, 10 min and 15 min respectively. The topograph of the MAO film surface and the film-substrate interface was observed by a scanning electron microscopy (SEM), and the composition was analyzed by an energy dispersive spectroscope (EDS) incorporated in the SEM. The phase and the microstructure of the film were analyzed by X-ray diffraction (XRD). The roughness of the film was measured using a roughness tester. The surface static contact angle was detected by a contact angle measurement instrument and the surface energy was calculated accordingly.

RESULTSWith the increase of processing time from 1 min to 15 min, the pore size increased from (1.30 ± 0.07) µm to (1.55 ± 0.09) µm, and film thickness increased from (10.2 ± 1.1) µm to (20.9 ± 2.9) µm. The content of the Ca in the film increased accordingly, and Ca/P increased from 1.99 to 2.45, and the surface energy increased from 24.62 mJ/m(2) to 39.49 mJ/m(2). Meanwhile, the XRD pattern indicated that rutile increased but anatase and titanium decreased gradually. At the time of 15 min, part of the MAO film peeled off.

CONCLUSIONSProcessing time has impact on the thickness, surface topography, crystal component and surface energy of titanium MAO coating. MAO film treated for 5 - 10 min demonstrated favorable surface properties.

Dental Implants ; Dental Materials ; chemistry ; Materials Testing ; Oxidation-Reduction ; Surface Properties ; Titanium ; chemistry

Since the first outbreak of avian influenza A(H7N9) virus in humans was identified in 2013, there have been five seasonal epidemics observed in China. An earlier start and a steep increase in the number of humans infected with H7N9 virus was observed between September and December 2016, raising great public concern in domestic and international societies. The epidemiological characteristics of the recently reported confirmed H7N9 cases were analysed. The results suggested that although more cases were reported recently, most cases in the fifth epidemic were still highly sporadically distributed without any epidemiology links; the main characteristics remained unchanged and the genetic characteristics of virus strains that were isolated in this epidemic remained similar to earlier epidemics. Interventions included live poultry market closures in several cities that reported more H7N9 cases recently.

In the spirit of curriculum-centered principle, the research group actively explored the content construction, operation mode and teaching method reform of comprehensive experimental course of basic medicine. Guided by diseases and application, experimental teaching courses based on the preparation and intervention process of animal models of diseases were extensively carried out, and "innovative" experimental items were added to ensure that the course contents reflected the frontier and the times. The course construction was guided by the training of biomedical frontier methods, making efforts to cultivate students' learning ability, practical ability and innovative ability. In addition, on the basis of optimizing the course content construction oriented to practical application and innovation, a virtual simulation experiment project is built, combined with online-offline hybrid teaching, to improve the comprehensive practical ability of trainees.

Background/Aims: Nonalcoholic steatohepatitis (NASH) is a progressive form of nonalcoholic fatty liver disease (NAFLD) characterized by hepatic steatosis, inflammation, hepatocellular injury, and fibrosis. We aimed to investigate the usefulness of a key biomarker, lipocalin-2 (LCN2), for the detection of NASH progression. Methods: A mouse NASH model was established using a high-fat diet and a high-sugar drinking water. Gene expression profile of the NASH model was analyzed using RNA sequencing. Moreover, 360 NAFLD patients (steatosis, 83; NASH, 277), 40 healthy individuals, and 87 patients with alcoholic fatty liver disease were recruited. Results: Inflammatory infiltration, focal necrosis in the leaflets, steatosis, and fibrosis were documented in the mouse liver. In total, 504 genes were differentially expressed in the livers of NASH mice, and showed significant functional enrichment in the inflammation-related category. Upregulated liver LCN2 was found to be significantly interactive with various interleukins and toll-like receptors. Serum LCN2 levels were significantly increased in NAFLD patients. Serum LCN2 levels were correlated with steatosis, intralobular inflammation, semiquantitative fibrosis score, and nonalcoholic fatty liver disease activity score. The area under the curve of serum LCN2 was 0.987 with a specificity of 100% and a sensitivity of 93.5% for NASH diagnosis, and 0.977 with almost the same specificity and sensitivity for steatosis. Conclusions LCN2 might be involved in the transition from NAFL to NASH by mediating inflammation. Serum LCN2 levels might be a novel biomarker for the diagnosis of NASH.

Background/Aims: Nonalcoholic steatohepatitis (NASH) is a progressive form of nonalcoholic fatty liver disease (NAFLD) characterized by hepatic steatosis, inflammation, hepatocellular injury, and fibrosis. We aimed to investigate the usefulness of a key biomarker, lipocalin-2 (LCN2), for the detection of NASH progression. Methods: A mouse NASH model was established using a high-fat diet and a high-sugar drinking water. Gene expression profile of the NASH model was analyzed using RNA sequencing. Moreover, 360 NAFLD patients (steatosis, 83; NASH, 277), 40 healthy individuals, and 87 patients with alcoholic fatty liver disease were recruited. Results: Inflammatory infiltration, focal necrosis in the leaflets, steatosis, and fibrosis were documented in the mouse liver. In total, 504 genes were differentially expressed in the livers of NASH mice, and showed significant functional enrichment in the inflammation-related category. Upregulated liver LCN2 was found to be significantly interactive with various interleukins and toll-like receptors. Serum LCN2 levels were significantly increased in NAFLD patients. Serum LCN2 levels were correlated with steatosis, intralobular inflammation, semiquantitative fibrosis score, and nonalcoholic fatty liver disease activity score. The area under the curve of serum LCN2 was 0.987 with a specificity of 100% and a sensitivity of 93.5% for NASH diagnosis, and 0.977 with almost the same specificity and sensitivity for steatosis. Conclusions LCN2 might be involved in the transition from NAFL to NASH by mediating inflammation. Serum LCN2 levels might be a novel biomarker for the diagnosis of NASH.

OBJECTIVETo investigate the time course of calpain activity changes in rat neurons following fluid percussion injury (FPI) under normothermia (37 degrees celsius;) and mild hypothermia (32-/+0.5) degrees celsius;.

METHODSIn vitro cultured rat neurons were subjected to FPI followed by application of mild hypothermia for intervention at different time points, and the changes in intraneuronal calpain activity following FPI and the interventional effect of mild hypothermia on calpain activity were evaluated by UV-spectrophotometry at different time points.

RESULTSRemarkable changes occurred in calpain activity in the neurons following FPI at 37 degrees celsius;, and mild hypothermia produced obvious interventional effect on calpain activity in close relation to the timing of intervention initiation.

CONCLUSIONIntraneuronal calpain activity changes following FPI are involved in the pathological process of cellular injury, and mild hypothermia might offer protection against traumatic brain injury to some extent by regulating calpain activity. The interventional effect of mild hypothermia is associated with the timing of the intervention initiation.

Animals ; Calpain ; metabolism ; Female ; Hypothermia, Induced ; Neurons ; metabolism ; pathology ; Percussion ; Pregnancy ; Rats ; Rats, Wistar ; Time Factors

OBJECTIVETo investigate the role of specificity protein 1 (Sp1) in regulating radiosensitivity of cervical cancer cell lines.

METHODSWe analyzed Sp1 expression in 6 different cervical cancer cell lines (SiHa, HeLa, Caski, Me180, Ms751, and C33a) using Western blotting and real-time PCR. Clonogenic survival assay and curve fitting were used to assess the changes in radiosensitivity of Me180 cells transfected with lentivirus-mediated shRNA vector targeting sp1 and HeLa cells transfected with sp1 over-expression vector.

RESULTSIn the 6 cell lines tested, the cellular expression levels of Sp1 decreased gradually in the order of Me180, Caski, C33a, SiHa, Ms751, and HeLa. SP1 knockdown with lentivirus-mediated shRNA significantly lowered the survival rate of Me180 cells following radiation exposure (P<0.05), and obviously lowered the values of SF2, D0 and Dq but significantly increased α/β of the cells. Compared with the cells transfected with the mock vector, HeLa cells with sp1 over-expression showed a significantly increased survival following radiation exposure (P<0.05) with obviously increased values of SF2, D0 and Dq but significantly lowered α/β.

CONCLUSIONSilencing Sp1 can increase the radiosensitivity while Sp1 overexpression enhances the radioresistance of cervical cancer cell lines, suggesting an important role of Sp1 in radiotherapy for cervical cancer.

BACKGROUNDStroke occurs upon obstruction of cerebral blood circulation and is clinically characterized by sudden onset symptoms. Advanced age is the main risk factor of stroke, but cases of pediatric stroke have been rarely reported. This study aimed to determine the etiology, clinical presentation, and radiologic features of neurological deficit for pediatric arterial ischemic stroke (PAIS).

METHODSThe medical records of 42 PAIS patients (age range: 9 months to 13 years) treated at Wuhan Children's Hospital between July 2007 and January 2011 were retrospectively reviewed. Infarction location was first determined by craniocerebral computed tomography and magnetic resonance (MR) imaging. The stenotic or occluded main cerebral arteries and/or branches were determined by MR angiography and digital subtraction angiography.

RESULTSThe majority of the 42 PAIS cases (66.7%, n = 28) were ≤ 3 years old (vs. >3 years old: 33.3%, n = 14; P<0.05), but the male: female ratio was similar in both groups (P > 0.05). The most frequently reported signs and symptoms for both age groups were limited physical activity followed by convulsions and delirium, but convulsions were more prevalent in children ≤ 3 years-old. Children > 3 years-old mainly experienced the limited physical activity symptoms, including hemiparalysis, aphasia, and ataxia. For all 42 cases, the most frequent etiologies were infections (38.1%, n = 16), iron deficiency anemia (16.7%, n = 7), and moyamoya syndrome (11.9%, n = 5). The predominant infarcts among all cases were middle cerebral artery (63.6%, n = 21) and basal ganglia (64.3%, n = 27).

CONCLUSIONSPAIS occurs more frequently in younger children and this group most frequently presents with convulsion as the initial symptom. The overall etiologies of PAIS may be different from those of adult stroke and the involved regions may be distinguishing features of PAIS or its different forms, but more research is required.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Radiography ; Retrospective Studies ; Stroke ; diagnosis ; diagnostic imaging ; etiology

Lynch syndrome (LS), an autosomal dominantly inherited disease previously known as hereditary non-polyposis colorectal cancer (HNPCC), leads to a high risk of colorectal cancer (CRC) as well as malignancy at certain sites including endometrium, ovary, stomach, and small bowel (Hampel et al., 2008; Lynch et al., 2009). Clinically, LS is considered the most common hereditary CRC-predisposing syndrome, accounting for about 3% of all CRC cases (Popat et al., 2005). LS is associated with mutations of DNA mismatch repair (MMR) genes such as MLH1, MSH2, MSH6, PMS2, and EPCAM (Ligtenberg et al., 2009; Lynch et al., 2009), which can trigger a high frequency of replication errors in both microsatellite regions and repetitive sequences in the coding regions of various cancer-related genes. Immunohistochemistry (IHC) tests followed by genetic analysis of these mutations play a significant role in diagnosis, treatment determination, and therapeutic response prediction of LS (Lynch et al., 2009; Alex et al., 2017; Ryan et al., 2017). Here, we report substitution of one base-pair in exon 1 of MLH3 (c.1397C>A) and a frameshift mutation in exon 19 of MLH1 (c.2250_2251ins AA) in a 43-year-old Chinese male with an LS pedigree.
Adult ; Asian People/genetics* ; China ; Colorectal Neoplasms, Hereditary Nonpolyposis/genetics* ; Exons ; Female ; Frameshift Mutation ; Germ-Line Mutation ; Humans ; Male ; MutL Protein Homolog 1/genetics* ; MutL Proteins/genetics* ; Pedigree