Objective To measure the concentration of serum transthyretin (TTR) of patients with different stages of diabetic retinopathy (DR).Methods A total of 176 patients with diabetes mellitus were included in this study.There were 104 males and 72 females.The patients aged from 21 to 74 years,with the mean age of(56± 11) years.The diabetes duration raged from 1 to 30 years,with the mean diabetes duration of (10 ± 7) years.The HbA 1C was 5.2％-14.1％,with the mean HbA 1C of (8.6 ± 2.0)％.According to the fundus examination,58 patients had DR (33.0％),but the other 118 patients not (67.0％).For these DR patients,10 patients were in stage Ⅰ (5.7％),26 patients in stage Ⅱ (14.8％),8 patients in stage Ⅲ (4.5％),and 14 patients in stage Ⅳ (8.0％).The concentration of serum TTR was measured by enzyme-linked immunosorbentassay kit.The differences in the concentration of serum TTR between different DR stages were compared.Bivariate analysis was used to analyze the influencing factors of TTR.Results The concentrations of serum TTR of the patients without DR or with DR of stage Ⅰ to Ⅳ were (224.96±65.47),(383.68± 102.99),(247.44±63.21),(228.2 ± 45.89),(189.34± 70.12) mg/L,respectively.The difference between different DR stages was statistically significant (F=14.690,P＜ 0.001).Bivariate analysis showed that the concentration of TTR was correlation to DR (r=0.179,P=0.017).There was no correlation between the concentration of TTR and diabetes duration (r=-0.027,P=0.727),hypertension (r=0.018,P=0.810),hyperlipoidemia (r=0.101,P=0.182),and the use of insulin (r=-0.032,P=0.675).Conclusion The concentration of serum TTR was increased in early DR patients,and gradually decreased with the progression of DR.The concentration of TTR is correlated to DR.

The present study was aimed to investigate the damage effect of hepatocyte growrh promoting substance (HGS) on the HL-60 cell DNA in vitro and to explore the possible mechanism underlying the effect. Experiment was divided into 3 groups: one was control group, in which 0.9% NaCl solution was added, and other two were experimental group 1 and group 2, where 22.5 microg/ml and 40 microg/ml HGS were added, respectively. HL-60 cell growths were compared between groups with and without HGS. Single cell gel electrophoresis (SCGE) was used to detect DNA damage of HL-60 cell. DNA electrophoresis was used to detect the apoptosis of HL-60 cells caused by HGS. The results showed that the inhibitory effects of HGS on growth of HL-60 cells were observed in group with 22.5 microg/ml and group with 40 microg/ml after culture for 2 days, the DNA ladder and the apoptosis of HL-60 cells occurred in these 2 groups on day 2 after addition of HGS, the counts of HL-60 cells with comet tail in these experimental groups were found to be more increased in comparison with control group. In conclusion, the HGS can inhibit the growth of HL-60 cell and the apoptosis of HL-60 cells should be induced through pathway of DNA damage caused by HGS.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; genetics ; Cattle ; Cell Proliferation ; drug effects ; Comet Assay ; DNA Damage ; DNA, Neoplasm ; analysis ; genetics ; HL-60 Cells ; Humans ; Peptides ; pharmacology

OBJECTIVETo investigate whether Ca2+ contribute to cardiomyocyte hypertrophy induced by tumor necrosis factor-alpha (TNF-alpha) through PI3-kinase pathway.

METHODSThe protein content was assayed with Lowry's method. The cardiomyocytes volumes were measured by computer photograph analysis system. The protein synthesis was assayed with [3H]-leucine incorporation method. [Ca2+]i transient was measured by Till image system by cell-loading Fura-2/AM.

RESULTS(1) TNF-alpha significantly induced the increase of protein content, [3H]-leucine incorporation and cell size. These responses were significantly suppressed by LY294002, a selective PI3-kinase inhibitor. Verapamil, L-type calcium channels antagonist, slightly attenuated TNF-alpha-induced these responses. (2) TNF-alpha increased the amplitude of the spontaneous Ca2+ transients in cultured ventricular myocytes from the neonatal rat; PI3-kinase inhibitor LY294002 could suppress the elevation induced by TNF-alpha, but calcium antagonist verapamil took the minor effects of TNF-alpha on [Ca2+]i metabolism.

CONCLUSIONIncreasing the intercellular free Ca2+ level may play an essential role in TNF-alpha-induced cardiomyocyte hypertrophy through PI3-kinase pathway in rats, while L-type calcium channel takes the minor effects on it.

Animals ; Calcium ; metabolism ; Calcium Channels, L-Type ; metabolism ; Chromones ; pharmacology ; Female ; Hypertrophy ; Male ; Morpholines ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo investigate the dynamic changes of myocardial matrix metalloproteinases (MMPs) activities in mice with viral myocarditis (VM) and their relationships with cardiac function and myocardial collagen amount and to explore the role of MMPs in the pathologic lesion of VM.

METHODSSixty-five six-week-old male DBA/2 mice were obtained from the Chinese Academy of Medical Sciences. They were divided into two groups randomly. Mice in infected group (n=50) were inoculated intraperitoneally with 0.14 ml of coxsackievirus B3 (CVB3, Nancy strain). Control mice (n=15) were inoculated intraperitoneally with 0.14 ml of Eagle's solution. Eight infected mice were sacrificed on day 3, 7, 10, 21 and 30, respectively and fifteen control mice were killed on day 30 after inoculation. Total protein concentration was determined according to the method of Bradford, while MMPs activities were measured with SDS-PAGE type substrate gels embedded with type I gelatin (zymography). Echocardiographic studies were performed under anesthesia with 3% chloralhydrate intraperitoneally (0.01-0.015 ml/g). Cardiac systolic function indexes, such as peak velocity of aorta (Vp) and flow velocity integral of aorta (Vi) were determined by echocardiography. Histological cross sections of hearts were stained with hematoxylin-eosin and myocardial histopathologic scores were counted under optical microscope. Myocardial collagen amount was measured by determination of hydroxyproline quantification.

RESULTSIn virus-infected mice, both MMP-2 and MMP-9 activities were increased significantly compared with those in controls and reached the peak on day 10 (P < 0.01). On day 10, cardiac systolic function indexes (Vp and Vi) were all significantly lower than those at other stages after virus inoculation and in control group (P < 0.05). There was no obvious elevation in myocardial collagen amount in mice with VM at acute stage (P > 0.05). While the myocardial collagen amount in infected group at recovery stage (on day 21 and 30) increased significantly compared with controls. MMP-2 and MMP-9 activities positively correlated with myocardial histopathological scores, respectively (r =0.801, 0.821 P < 0.01), while they negatively correlated with Vp (r = -0.649, -0.683, P < 0.01) and Vi, respectively (r = -0.711, -0.755, P < 0.01). However, Vp and Vi negatively correlated with myocardial histopathological scores (r = -0.756, -0.584, P < 0.01).

CONCLUSIONSIn mice with VM, the activities of myocardial MMP-2 and MMP-9 at acute stage increased significantly, then myocardial collagen amount elevated in recovery stage. These changes were associated with myocardial remodeling and cardiac dysfunction. Myocardial MMP activities are important markers of myocardial pathologic lesion. They are of value in the evaluation of the severity of myocardial damage and cardiac dysfunction in mice with VM.

Animals ; Collagen ; metabolism ; Coxsackievirus Infections ; complications ; Disease Models, Animal ; Echocardiography ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinases ; metabolism ; Mice ; Mice, Inbred DBA ; Myocarditis ; diagnostic imaging ; pathology ; physiopathology ; virology ; Myocardium ; metabolism ; pathology ; Systole ; Ventricular Dysfunction ; diagnostic imaging ; physiopathology ; Ventricular Remodeling

Objective To explore the source of human infection H9N2 avian influenza virus (AIV). Methods Environmental AIV nucleic acid monitoring was conducted for live poultry markets in Changsha city from 2014 to 2015, and data of human infection H9N2 subtype AIV cases worldwide were collected. Phylogenetic trees of hemagglutinin(HA), neuraminidase(NA)and non-structural protein(NS)genes from human infection H9N2 subtype AIV, the live poultry markets environmental H9N2 subtype AIV and partial avian H9N2 subtype AIV were constructed using the MEGA 6.06 software, respectively. Results In 2014-2015, H9 subtype AIV had the highest nucleic acid positive rate (44.76%) in the live poultry markets environment of Changsha city, and the pollution was serious. A total of 27 cases of human infection with H9N2 subtype AIV had been reported worldwide, and most of these patients recovered after treatments.Epidemiological survey showed that 59.26% (16/27) of cases had a clear history of exposure to poultry or live poultry markets. The phylogenetic trees of HA, NA and NS genes showed that the human infection H9N2 subtype AIV isolates isolated from Hunan and Guangdong were closely related to the H9N2 subtype AIV isolated from the live poultry markets environment in Hunan and Guangdong provinces from 2013 to 2016. The nucleotide similarity was as high as 97%-99%. Conclusion Live poultry market is one of the sources of human infection with H9N2 influenza virus.

Objective To study the analytical sensitivity on 31 HBsAg enzyme immunoassy (EIA) test kits.Methotis Thirty one HBsAg EIA kits produced by domestic or overseas manufactories and applied for approval during May 2007 to May 2008,were evaluated using the national reference panels.The hyperbolic curve of the log A value and log concentration for the national sensitivity standards was established.The cut-off value of each kit was substituted into the curvilinear equation to determine the analytical sensitivity which was compared between different HBsAg EIA kits.Results Twenty seven(351 lots) domestic and 4(27 lots) overseas kits were compared.Among 378 lots of the 31 HBsAg EIA kits,only 2 lots of the domestic kits had a lower sensitivity when tested with the national HBsAg reference panels,with an average approvalr ate of 99.43%(349/351).The mean analytical sensitivity of the domestic kits for adr,adw,ay serotypes were 0.307,0.419,0.513 ng/ml,respectively.There was a significant difierence between serotypes (F=97.30,P<0.01).The mean analytical sensitivity of the overseas kits for adr,adw,ay serotypes were 0.054,0.066,0.050 ng/ml respectively,with no significant difference between serotypes(F=0.65,P>0.05).The analytical sensitivity of the overseas kits for all the three serotypes was higher than that of the domestic kits(P<0.01).There was no significant difference found between the analytical sensitivities of the kits produced by the same manufactory using 30- or 60- minute incubation of detection(P>0.05).In contrast,there was significant diffefence noticed between the analytical sensitivities of the kits produced by the same manufactory when tested for 10 or 15- minute coloration of the results(P<0.01).Conclusion Analytical sensitivity of the HBsAg EIA domestic kits should be further improved,especiatry for detecting adw and ay serotypes.

BACKGROUNDMatrix metalloproteinases (MMPs) are the major regulators of collagen degradation involved in the pathogenesis of several diseases of the heart. The purpose of this study was to investigate the dynamic changes in myocardial MMP activity in mice with viral myocarditis (VM), the relationship between MMP activity and both cardiac function and the quantity of myocardial collagen, and the role MMPs playing in the pathological lesions of VM.

METHODSSixty-five six-week-old male DBA/2 mice were divided into two groups. Mice in the infected group (n = 50) were inoculated intraperitoneally with 0.14 ml of Coxsackievirus B3 (CVB3, Nancy strain). Control mice (n = 15) were inoculated intraperitoneally with 0.14 ml of Eagle's medium. Eight infected mice and three control mice were sacrificed on each of days 3, 7, 10, 21 and 30 after inoculation. MMP activity was measured on an SDS-PAGE substrate gel embedded with type I gelatin (zymography). Echocardiographic studies were performed under anesthesia with 3% chloralhydrate administered intraperitoneally (0.01 ml/g - 0.015 ml/g). Cardiac systolic function indices, such as peak velocity of the aorta (Vp), flow velocity integral of the aorta (Vi), ejection fraction (EF), and fractional shortening (FS) were determined by echocardiography. Histological cross sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope. The amount of myocardial collagen was measured by means of hydroxyproline quantification.

RESULTSIn virus-infected mice, both MMP-2 and MMP-9 activities were significantly higher than in control mice, reaching a peak on day 10 (P < 0.01). On day 10, cardiac systolic function indices (EF, FS, Vp, and Vi) were all significantly lower compared both to other stages following viral inoculation and to the control group (P < 0.05). In the acute stage, the amount of myocardial collagen in mice with VM was not significantly different from normal control mice (P > 0.05). However, the amount of myocardial collagen in infected mice at the recovery stage (on days 21 and 30) was significantly greater than those of the control mice. MMP-2 and MMP-9 activities positively correlated with myocardial histopathological scores (r = 0.801, 0.821, P < 0.01) and negatively correlated with Vp (r = -0.649, -0.683, P < 0.01) and Vi (r = -0.711, -0.755, P < 0.01). However, Vp negatively correlated with myocardial histopathological scores (r = -0.756, P < 0.01).

CONCLUSIONSIn mice with VM, the activities of myocardial MMP-2 and MMP-9 increase significantly during the acute stage, and the total quantity of myocardial collagen increases by the time of recovery. These changes are associated with myocardial interstition remodeling and cardiac dysfunction. MMP activity is an important reference marker for myocardial pathological lesions and can be used to evaluate the severity of myocardial interstitial damage and cardiac dysfunction.

Animals ; Collagen ; analysis ; Enterovirus B, Human ; Enterovirus Infections ; enzymology ; pathology ; physiopathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred DBA ; Myocarditis ; enzymology ; pathology ; physiopathology

Intestinal dysbiosis and disrupted bile acid(BA)homeostasis are associated with obesity,but the precise mechanisms remain insufficiently explored.Hepatic protein phosphatase 1 regulatory subunit 3G(PPP1R3G)plays a pivotal role in regulating glycolipid metabolism;nevertheless,its obesity-combatting potency remains unclear.In this study,a substantial reduction was observed in serum PPP1R3G levels in high-body mass index(BMI)and high-fat diet(HFD)-exposed mice,establishing a positive correlation between PPP1R3G and non-12α-hydroxylated(non-12-OH)BA content.Additionally,hepatocyte-specific overexpression of Ppp1r3g(PPP1R3G HOE)mitigated HFD-induced obesity as evidenced by reduced weight,fat mass,and an improved serum lipid profile;hepatic steatosis alleviation was confirmed by normalized liver enzymes and histology.PPP1R3G HOE considerably impacted systemic BA homeostasis,which notably increased the non-12-OH BAs ratio,particularly lithocholic acid(LCA).16S ribosomal DNA(16S rDNA)sequencing assay indicated that PPP1R3G HOE reversed HFD-induced gut dysbiosis by reducing the Firmicutes/Bacteroidetes ratio and Lactobacillus population,and elevating the relative abundance of Blautia,which exhibited a positive correlation with serum LCA levels.A fecal microbiome transplantation test confirmed that the anti-obesity effect of hepatic PPP1R3G was gut microbiota-dependent.Mechanistically,PPP1R3G HOE markedly suppressed hepatic cholesterol 7α-hydroxylase(CYP7A1)and sterol-12α-hydroxylase(CYP8B1),and concurrently upregulated oxysterol 7-α hydroxylase and Takeda G protein-coupled BA receptor 5(TGR5)expression under HFD conditions.Furthermore,LCA administration significantly mitigated the HFD-induced obesity phenotype and elevated non-12-OH BA levels.These findings emphasize the significance of hepatic PPP1R3G in ameliorating diet-induced adiposity and hepatic steatosis through the gut microbiota-BA axis,which may serve as potential ther-apeutic targets for obesity-related disorders.

This article reports the clinical and genetic features of two cases of cerebral creatine deficiency syndrome I (CCDSI) caused by SLC6A8 gene mutations. Both children were boys. Boy 1 (aged 2 years and 10 months) and Boy 2 (aged 8 years and 11 months) had the clinical manifestations of delayed mental and motor development, and convulsion. Their older brothers had the same symptoms. The mother of the boy 1 had mild intellectual disability. The genetic analysis showed two novel homozygous mutations, c.200G>A(p.Gly67Asp) and c.626_627delCT(p.Pro209Argfs*87), in the SLC6A8 gene on the X chromosome, both of which came from their mothers. These two novel mutations were rated as possible pathogenic mutations and were not reported in the literature before. This study expands the mutation spectrum of the SLC6A8 gene and has great significance in the diagnosis of boys with delayed development, and epilepsy.
Child ; Child, Preschool ; Creatine ; Epilepsy ; Genetic Testing ; Humans ; Male ; Mutation ; Nerve Tissue Proteins ; genetics ; Plasma Membrane Neurotransmitter Transport Proteins ; genetics ; Syndrome