AIM:To explore the expression of anoctamin 1 (ANO1), one of calcium-activated chloride chan-nels ( CaCCs) , in mouse cardiomyocytes and its functional properties.METHODS:The cardiomyocytes from the myocar-dial tissues of C57BL/6 mice were isolated with enzyme and purified by the differential adherent method.The cells were stained with monoclonal anti-sarcomeric actin and Cy3 to evaluate the purity of the myocardial cells.RT-PCR was used to detect the mRNA expression of ANO1 in the mouse cardiomyocytes.The protein expression of ANO1 in the mouse cardio-myocytes was determined by Western blotting analysis.The fluorescence quenching kinetics experiment was used to identify the ion transport properties of ANO1 in the mouse cardiomyocytes.RESULTS: The results of RT-PCR confirmed that ANO1 was expressed in freshly isolated myocardial cells.The results of Western blotting clearly demonstrated the protein expression of ANO1 in primarily cultured myocardial cells.Fluorescence quenching kinetics experiment on freshly isolated single myocardial cell revealed a pronounced outward rectifying property of the ANO1.The functional properties were simi-lar to the classic CaCCs.CONCLUSION:ANO1 expression was identified in the mouse myocardial cells.The function of CaCCs was generated by ANO1, suggesting that ANO1 is the molecular basis of CaCCs.

We wished to assess the role of chlamydia micro virus capsid protein Vp3 in recombinant molecules, chart its molecular evolution, screen the wild-type strain, and reveal its value in clinical research. Using a protein BLAST multiple-alignment program, we compared various strains of Chlamydia micro virus capsid protein Vp3 sequences. Using a "distance tree" of those results, we created a phylogenetic tree. We applied the Karplus-Schulz method of flexible-region analyses for highly conserved alignments of amino-acid sequences. Gamier-Robson and Chou-Fasman methods were employed to analyze two-level structures of sequences. The Emini method was used for analyses of the accessibility of surface epitopes. Studies of hydrophilic proteins were undertaken using Kyte-Doolittle and Hopp-Woods methods. Analyses of antigen epitopes helped to reveal the antigen index using the Jameson-Wolf method. All sequences in the six strains of chlamydia micro virus capsid protein Vp3 were highly conserved, with the main differences being between Vp3 protein in Chp1 and the other five strains of the micro virus. The viral strain of Vp3 protein was based mainly on micro-alpha helix structures, and multiple epitopes were noted in highly conserved regions. Vp3 protein was highly conserved structurally, and was an important protein of the chlamydiaphage capsid. Vp3 protein has a complicated molecular structure, highly conserved regions with strong immunogenicity, and has considerable research value.
Amino Acid Sequence ; Capsid Proteins ; chemistry ; genetics ; immunology ; Chlamydia ; genetics ; immunology ; Conserved Sequence ; Epitope Mapping ; Evolution, Molecular ; Molecular Sequence Data ; Recombination, Genetic

The fibronectin was purified from human plasma by Gelatin-Sepharose 4B affinity chromatography. The rabbits were immunized with the fibronectin. the raw anti-fibronectin sera were absorbed bythe solid phase of the fibronectin free human serum. Then the specific anti-fibronectin sera were obtained. The fibronectins deposited on human bullet wounds, incised wounds as well as on blunt forceinJuries were demonst.ated by PAP immunohistochemical study using diluted anti-fibronectin serum.The results were quite sati factory.

OBJECTIVETo analyze the mutations in a pedigree with maternally inherited sensorineural hearing loss, and to investigate whether 235delC heterozygote mutation in gap junction protein beta 2 (GJB2) gene modulates the severity of hearing loss associated with the A1555G mitochondrial mutation.

METHODSThe PCR products were digested with the Alw26 I restriction enzyme, followed by direct sequencing to detect the mitochondrial mutations in 72 members of a core pedigree of an extensive family with matrilineal nonsyndromic deafness; 235delC mutation of the GJB2 gene was screened in this family by using the Apa I restriction enzyme and direct sequencing.

RESULTSThe A1555G mutation of the mitochondrial DNA was present in all 27 members of maternal line, out of them, 21 members had phenotype of deafness (77.8%), with a high penetrance. Only three maternal line members of 72 members possessed 235delC heterozygote mutations, and the three had different phenotypes.

CONCLUSIONThe A1555G homozygous mutation of mitochondrial DNA is the susceptive etiological factor of nonsyndromic deafness in this family, but in the study of this pedigree, the 235delC heterozygous mutation in GJB2 gene may not aggravate the symptoms of hearing loss associated with the A1555G mitochondrial mutation.

Base Sequence ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Female ; Hearing Loss, Sensorineural ; genetics ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction

Objective To study the cytomorphologic features of anaplastic lymphoma kinase ( ALK )-rearranged pulmonary adenocarcinoma.Methods The morphologic features in 153 pulmonary adenocarcinoma cytology specimens encountered during the period from September, 2011 to April, 2015 in Shanghai Cancer Hospital were retrospectively reviewed.Fluorescence in-situ hybridization ( FISH) and/or immunohistochemistry ( Ventana D5F3) for ALK gene rearrangement were carried out.The samples studied included 34 pleural effusion specimens, 40 endobronchial ultrasound-guided transbronchial needle aspirates ( EBUS-TBNA) and 79 fine needle aspirates of palpable masses on body surface.Results Thirty-nine cases (25.5%) of ALK-rearranged samples were identified by FISH and/or immunohistochemistry, including 3 cases diagnosed by FISH and 36 cases by both technologies.The median age of the ALK-positive group was 50 years, significantly younger than that of the ALK-negative group (60 years old,P=0.002) .Only 4 of the ALK-positive patients were smokers, which was significantly less than that of the ALK-negative group (P<0.01).In ALK-positive group, 3 cases showed cribriform pattern with prominent nucleoli, 3 cases showed cribriform pattern with mucin-rich cells and 8 cases showed extracellular mucus with mucin-rich cells.The above cytomorphologic patterns were significantly less common in ALK-negative tumors ( P <0.01 ) . Conclusions ALK-rearranged lung adenocarcinoma is associated with certain distinctive morphologic patterns, including cribriform architecture, presence of prominent nucleoli, mucin-rich cells and extracellular mucus, which can be observed in cytology specimens ( including conventional smears and cell block sections) .These findings, when combined with clinical features, may give clues to detection of ALK-positive cases.

Objective:Analyzing the relationship between negative life events and depressive symptoms in university freshmen,and the mediating effects of neuroticism and the moderating role of exercise frequency.Meth-ods:A sampling of 8 079 university freshmen,and the Patient Health Questionnaire was used to assess depressive symptoms,the Eysenck Personality Inventory-Neuroticism subscale to assess neuroticism,the self-administered questionnaire to assess the number of negative life events that the participants had experienced and the exercise fre-quency.Model 4 in the Process plug-in was used to test the mediating effect of neuroticism,and Model 7 to test the moderating role of exercise frequency.Results:The numbers of negative life events were positively correlated with the depressive symptoms scores(r=0.16,P＜0.01),and were positively correlated with the neuroticism scores(r=0.26,P＜0.01).The neuroticism scores were positively correlated with the depressive symptoms scores(r=0.52,P＜0.01).Neuroticism score partially mediated between negative life events and depressive symptoms score,with a mediating effect of 78.4％,and exercise frequency score moderated between negative life events and neuroti-cism scores(β=-0.05,P=0.032).Conclusion:Negative life events are associated with depressive symptoms,neuroticism plays a mediating role,and exercise frequency could moderate negative life events and neuroticism.

OBJECTIVETo study the cytomorphologic features of anaplastic lymphoma kinase (ALK)-rearranged pulmonary adenocarcinoma.

METHODSThe morphologic features in 153 pulmonary adenocarcinoma cytology specimens encountered during the period from September, 2011 to April, 2015 in Shanghai Cancer Hospital were retrospectively reviewed. Fluorescence in-situ hybridization (FISH) and/or immunohistochemistry (Ventana D5F3) for ALK gene rearrangement were carried out. The samples studied included 34 pleural effusion specimens, 40 endobronchial ultrasound-guided transbronchial needle aspirates (EBUS-TBNA) and 79 fine needle aspirates of palpable masses on body surface.

RESULTSThirty-nine cases (25.5%) of ALK-rearranged samples were identified by FISH and/or immunohistochemistry, including 3 cases diagnosed by FISH and 36 cases by both technologies. The median age of the ALK-positive group was 50 years, significantly younger than that of the ALK-negative group (60 years old, P = 0.002). Only 4 of the ALK-positive patients were smokers, which was significantly less than that of the ALK-negative group (P < 0.01). In ALK-positive group, 3 cases showed cribriform pattern with prominent nucleoli, 3 cases showed cribriform pattern with mucin-rich cells and 8 cases showed extracellular mucus with mucin-rich cells. The above cytomorphologic patterns were significantly less common in ALK-negative tumors (P < 0.01).

CONCLUSIONSALK-rearranged lung adenocarcinoma is associated with certain distinctive morphologic patterns, including cribriform architecture, presence of prominent nucleoli, mucin-rich cells and extracellular mucus, which can be observed in cytology specimens (including conventional smears and cell block sections). These findings, when combined with clinical features, may give clues to detection of ALK-positive cases.

Adenocarcinoma ; genetics ; pathology ; Biopsy, Fine-Needle ; China ; Gene Rearrangement ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; pathology ; Receptor Protein-Tyrosine Kinases ; genetics ; Retrospective Studies