Objective:To evaluate the efficiency of using artificial intelligence reading label system in diabetic retinopathy (DR) grading training among junior ophthalmologists and medical students.Methods:520 diabetic fundus images were randomly divided into 8 groups with 65 images in each group. 13 junior ophthalmologists and medical students were selected as the research objects. Each of them read 8 groups of pictures and evaluated the DR grading of each fundus image. The sensitivity, specificity and diagnostic test consistency (Q-kappa value) of grading results were analyzed with the DR grading given by 3 senior ophthalmologists as the gold standard. The average Q-kappa values of 13 subjects were compared between the first four times and the last four times.Results:Through 8 round reading, the average Q-kappa was elevated from 0.67 to 0.81. Average Q-kappa of round 1 to 4 was 0.77, and average Q-kappa of round 5 to 8 was 0.81. The participants were divided into two groups. Participants in group 1 were junior ophthalmologists and participants in group 2 were medical students. Average Q-kappa of group 1 was elevated from 0.71 to 0.76. Average Q-kappa of group 2 was elevated from 0.63 to 0.84.Conclusions:The artificial intelligence reading label system was a useful tool in training junior ophthalmologists and medical students in doing diabetic retinopathy grading.

Objective To establish the reference intervals of hemoglobin A 1c( HbA1c ) and fasting plasma glucose ( FPG ) in the first and second trimester of pregnancy in Chongqing , and to evaluate the viability of the combination of HbA 1c and FPG in screening gestational diabetes mellitus (GDM).Methods The study retrospectively selected the pregnant women seen at the Department of Obstetrics and Gynecology in Daping Hospital between September 2014 and August 2015.The results of FPG during 10-13 pregnant weeks and 75 g oral glucose tolerance test ( OGTT ) and HbA1c during 24-28 pregnant weeks were available.Totally 185 cases were assigned into GDM group , and 269 cases were assigned into normal group based on the American Diabetes Association ( ADA) guidelines.Reference intervals of HbA 1c and FPG in normal pregnant woman were developed .The difference of HbA 1c , FPG and OGTT results between two groups was analyzed.T-student test, NcMemar test,signed rank sum test, ROC curve were used for statistical analysis.Results The reference intervals of HbA 1c and FPG in first and second trimester were 4.58%-5.52%,4.21-5.49 mmol/L and 4.03-5.08 mmol/L.The FPG level in first and second trimester and HbA 1c level in GDM group vs normal group were(5.06 ±0.37) vs(4.85 ±0.32)mmol/L(t=6.569,P=0.000), 5.23(5.11,5.4) vs 4.74(4.54,4.91) mmol/L(z=-14.31,P=0.000)and 5.3(5.1,5.4)% vs 5.2(5.0, 5.3)%( z=-5.79,P=0.000) respectively.The area under receiver operating characteristic curve ( ROC) of HbA1c , and FPG in first and second trimester was 0.655, 0.659 and 0.890 respectively.When the cut-off value of HbA1c was 5.35%, the AUC of the combination of HbA 1c and FPG in second trimester was 0.898, the sensitivity was 0.838,and the specificity was 0.859.The kappa coefficient for identifying GDM between OGTT and the combined method was 0.692(P=0.000).Conclusion HbA1c combined with FPG is of some value in screening GDM.

Objective To study effects of low molecular weight heparin on liver fibrosis and the serum levels of TGF-?_1 in patients with chronic hepatitis B virus.Methods 45 patients with chronic hepatitis B virus were randomized into control group(the routine strategy)and trial group(the routine strategy + low molecular weight heparin).The period of treatment is 3 weeks.Serum hepatic fibrosis indices before and after heparin treatment were examined by RIA,the levels of serum TGF-?_1 before and after heparin treatment were examined by ELISA.Results Hepatic functions became significantly better in trial groups,serum PⅢP and type IV collage levels and the levels of serum TGF-?_1 in trial group decreased significantly after treatment.Conclusions The mechanism of anti-fibrosis action of LMWH may inhibited production of TGF-?_1 in patients with HBV.

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

AIMTo investigate the inhibition of amyloid beta-protein 42 (Abeta42) production in M146L cells by gamma-schisandrin.

METHODSM146L cells which can produce considerable Abeta42 in vitro were treated with gamma-schisandrin (1.67, 5.00 and 15.00 microg x mL(-1)), beta-secretase inhibitor (S4562, 100.00 microg x mL(-1)) and gamma-secretase inhibitor (S2188, 13.68 microg x mL(-1)), separately. Cell counting kit-8 (CCK-8) was used to assess cell viability. Enzyme-linked immunosorbent assay (ELISA) was carried out to determine the amount of Abeta42. Western blotting was used to examine C99, an intermediary product of APP cleaved by beta-secretase. beta-Secretase and gamma-secretase activities were assayed by commercial kits.

RESULTSThe CCK-8 assay indicated that different concentrations of gamma-schisandrin had no neurotoxicity on the cultured M146L. And the ELISA test showed that the amount of Abeta42 secreted by M146L cells treated with gamma-schisandrin (5.00 and 15.00 microg x mL(-1)) decreased obviously as compared with solvent control. The results of Western blotting test indicated that there was no change of C99 contents and beta-secretase activity in gamma-schisandrin treated cells, while gamma-secretase activity decreased obviously.

CONCLUSIONgamma-Schisandrin inhibited production of Abeta42 in M146L cells through inhibiting gamma-secretase.

Alzheimer Disease ; drug therapy ; Amyloid Precursor Protein Secretases ; antagonists & inhibitors ; metabolism ; Amyloid beta-Peptides ; antagonists & inhibitors ; biosynthesis ; Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Cyclooctanes ; Dose-Response Relationship, Drug ; Humans ; Lignans ; Peptide Fragments ; antagonists & inhibitors ; biosynthesis ; Polycyclic Compounds ; pharmacology

As a crucial signaling molecule, calcium plays a critical role in many physiological and pathological processes by regulating ion channel activity. Recently, one study resolved the structure of the transient receptor potential melastatin 2 (TRPM2) channel from Nematostella vectensis (nvTRPM2). This identified a calcium-binding site in the S2–S3 loop, while its effect on channel gating remains unclear. Here, we investigated the role of this calcium-binding site in both nvTRPM2 and human TRPM2 (hTRPM2) by mutagenesis and patch-clamp recording. Unlike hTRPM2, nvTRPM2 cannot be activated by calcium alone. Moreover, the inactivation rate of nvTRPM2 was decreased as intracellular calcium concentration was increased. In addition, our results showed that the four key residues in the calcium-binding site of S2–S3 loop have similar effects on the gating processes of nvTRPM2 and hTRPM2. Among them, the mutations at negatively charged residues (glutamate and aspartate) substantially decreased the currents of nvTRPM2 and hTRPM2. This suggests that these sites are essential for calcium-dependent channel gating. For the charge-neutralizing residues (glutamine and asparagine) in the calcium-binding site, our data showed that glutamine mutating to alanine or glutamate did not affect the channel activity, but glutamine mutating to lysine caused loss of function. Asparagine mutating to aspartate still remained functional, while asparagine mutating to alanine or lysine led to little channel activity. These results suggest that the side chain of glutamine has a less contribution to channel gating than does asparagine. However, our data indicated that both glutamine mutating to alanine or glutamate and asparagine mutating to aspartate accelerated the channel inactivation rate, suggesting that the calcium-binding site in the S2–S3 loop is important for calcium-dependent channel inactivation. Taken together, our results uncovered the effect of four key residues in the S2–S3 loop of TRPM2 on the TRPM2 gating process.

OBJECTIVETo analyze the sex chromosome meiotic segregation in inv(Y) patients by fluorescence in situ hybridization (FISH).

METHODSConventional cytogenetic procedures (GTG and CBG banding) and FISH were performed on metaphase chromosome. Three-color FISH was performed on sperm samples using a probe mixture containing CEPX, Tel Xp/Yp and Tel Xq/Yq to investigate the sex chromosome segregation of five inv(Y) (p11.1q11.2) carriers. A healthy man with normal semen parameters was used as control.

RESULTSThere was no statistical difference in the abnormal sex chromosome number and recombination frequencies in each spermatozoon from the patient in comparison with that in the control.

CONCLUSIONThere was no apparent sex chromosome abnormality in the sperm of the inv(Y) (p11.1q11.2) carriers. Sperm-FISH allows further understanding of the sex chromosome segregation pattern and an accurate genetic counseling.

Case-Control Studies ; Chromosome Inversion ; Chromosomes, Human, Y ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Meiosis ; genetics ; Recombination, Genetic ; Sex Chromosome Aberrations ; Spermatozoa ; metabolism ; pathology

OBJECTIVETo explore whether PI3K/Akt/mdm2 signalling pathway affect the sensitivity of gastric cancer cell line SGC7901 cells to doxorubicin.

METHODSGastric cancer cell line SGC7901 cells were exposed to doxorubicin and specific PI3K inhibitor wortmannin. Cell apoptosis was detected using flow cytometry. PI3K activity was detected by radioactive immunoprecipitation-kinase assay. Western blotting was employed to evaluate the expressions of PI3K-p85, pAkt-S473, Akt, pmdm2-S166 and p53.

RESULTSThe level of apoptosis in gastric cancer SGC7901 cells treated with doxorubicin was gradually increasing. wortmannin enhanced its effects significantly. PI3K activity and the expression of pAkt-S473 increased in a time-dependent manner, pmdm2-S166, p53 were also increased wortmannin inhibited phosphorylation of mdm2 and improved the p53 expression.

CONCLUSIONPI3K/Akt/mdm2 signalling pathway can be activated by doxorubicin and suppress apoptosis by promoting phosphorylation of mdm2. PI3K inhibitor wortmannin can enhance sensitivity of gastric cancer cells to chemotherapy.

Androstadienes ; pharmacology ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Enzyme Activation ; Humans ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Phosphorylation ; Protein Kinase Inhibitors ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-mdm2 ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

Objective To observe the expressing changes of apolipoprotein M (ApoM),tumor necrosis factor-α(TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in human retinal vascular endothelial cells (HRECs) under the high glucose culture condition and investigate the inhibitory effects of ApoM overepression on the expressions of TNF-α and MCP-1.Methods HRECs were cultured in DMEM containing 10％ fetal bovine serum and 5.5 mmol/L D-glucose and assigned to 6 groups.The cells in the normal control group were cultured in above culture medium;the cells in the high glucose group were treated using the DMEM with 30 mmol/L D-glucose;ApoM was transfected into the cells using lentiviral vector in the ApoM transfected group;lentiviral vector without ApoM sequence was transfected in the empty vector group;the cells transfected by empty vector were cultured in high glucose culture medium in the empty vector+high glucose group;the cells in the ApoM transfection+high glucose group were treated by ApoM sequence transfection and high glucose incubation.The relative expression of ApoM,TNF-α and MCP-1 mRNA was detected using real-time quantitative PCR,and the relative expression of ApoM protein was evaluated using Western blot assay.Results Compared with the normal control group,the mRNA expression levels of ApoM,TNF-α and MCP-1 in the high glucose group were significantly increased (t=5.517,3.295,2.555;all P＜0.05).HRECs grew well after infected with lentivirus.The relative expression level of ApoM mRNA in the ApoM transfected group was 236.400±39.270,which was significantly higher than 1.000±0.153 in the empty vector group (t=5.995,P＜0.01).An enhanced protein band of ApoM was seen in the ApoM transfected group,and the protein band was absent in the empty vector group.The relative expression band in the ApoM transfected group was 1.000± 0.249 and 2.978 ± 0.285 in the cells cultured with normal culture medium or high glucose culture medium,respectively,with a significant difference between them (t =5.056,P＜0.01).The relative expressions of TNF-α and MCP-1 in the mRNA levels were significantly different among the empty vector group,empty vector+high glucose group,ApoM transfected group and ApoM transfection + high glucose group (F =5.966,P =0.026;F =14.410,P =0.002).Compared with the empty vector+high glucose group,the relative expressions of TNF-α and MCP-1 mRNA were considerably reduced in the ApoM transfection+high glucose group (P=0.017,0.004).Conclusions High glucose environment up-regulates the expression of ApoM,MCP-1 and TNF-α in HRECs.Overexpression of ApoM inhibits the up-regulation of MCP-1 and TNF-α expression induced by high glucose.

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.