Objective To explore the association between the different forms of in vivo ghrelin—Acyl ghrelin( AG) ,Des-acyl ghrelin( DAG) and AG/DAG with insulin resistance( IR) in patients with type 2 diabetes mellitus. Methods From June 2017 to November 2017,eighty-three patients with type 2 diabetes hospitalized in (group T2DM) and 40 healthy subjects (group NC) were hospitalized in Jinling Clinical Medicine were selected. Height body mass,blood pressure,blood lipid,glycated hemoglobin ( HbA1c) and fasting blood glucose (FPG),fasting insulin (FINS),and fasting C peptide (F-C-p) were measured,and all subjects were left with fasting serum,and the concentration of AG and DAG were measured by enzyme linked immunosorbent assay (ELISA). The body mass index (BMI),total gastric starvation (T-ghrelin) level,AG/DAG,insulin resistance index ( HOMA-IR) , insulin sensitivity index ( HOMA-IS ) and islet beta cell function ( HOMA- beta ) were calculated. The differences of the above indexes between the two groups were compared, and the relationship between AG,DAG,T-ghrelin,AG/DAG and FPG,HOMA-IR,HOMA-IS and HOMA- beta in T2DM patients were analyzed. Results ( 1) There were no significant difference in SBP、DBP、TC、LDL-C、AG between group NC and group T2DM(P>0. 05). Compared with NC group,the age、TG、BMI、HbA1c、FPG、FINS、HOMA-IR、HOMA-β、AG/DAG were significantly higher in T2DM group ( t=2. 690,-1. 990, 0. 873, 14. 257, 10. 528, Z=2. 885,-3. 483,-2. 284;P<0. 01,P<0. 05) . The HDL-C,F-C-p,HOMA-IS,HOMA-beta,DAG and T-ghrelin in group T2DM were lower than those of NC group( t or Z=0. 477,-3. 812,-3. 395,-4. 4,-2. 916,-2. 834;P<0. 05 or P<0. 01) . ( 2) The correlation analysis showed that there was a positive correlation between AG and FPG in T2DM group (r=0. 252,P<0. 05),DAG and T-ghrelin were negatively correlated with HOMA-IR (r=-0. 394,-0. 384,P<0. 05),and AG/DAG was positively correlated with HOMA-IR (r=0. 394,0. 384,P<0. 05),but is negatively correlated with HOMA-IS (r=-0. 292,P<0. 05). (3) multivariate linear regression analysis showed that FPG in T2DM patients were the influencing factors of AG ( t=2. 865,P<0. 05) ,while FINS and BMI were the influencing factors of DAG( t=-2. 808、-0. 330,P<0. 05) andT-ghrelin( t=-2. 725、-0. 330, P<0. 05) . HOMA-IR and BMI are the influencing factors of AG/DAG ( t=3. 718,3. 069,P<0. 05) . Conclusion The levels of DAG and T-ghrelin in group T2DM were significantly lower than those in the normal population, and was negatively correlated with the insulin resistance index,and the ratio of AG/DAG was closely related to insulin resistance,and the level of AG was mainly affected by fasting blood glucose.

Objective To analyze horizontal transmission patterns of Streptococcus mutans among caries-active preschool children for early interventions of dental caries. Methods Plaque samples obtained from 20 caries-active preschool children between 4 and 5 years of age were cultured under anaerobic conditions for isolating S. mutans, which were identified by morphological and biochemical analyses and PCR using primers homologous to the surface protein glucosyltransferase B (gtfB). The genotypes of the isolated S. mutans strains were determined by arbitrarily primed PCR (AP-PCR). Results Of the 200 S. mutans isolates obtained, 19 were excluded by biochemical analysis, and the remaining 181 isolates were identified as S. mutans by PCR with primers of gtfB, showing 37 different genotypes as identified by AP-PCR. Six children were found to carry S. mutans of a single genotype, 11 carried 2 genotypes, 2 had 3 genotypes, and 1 had 4 genotypes;2 children from different classes were found to carry S. mutans of the same single genotype. Conclusion We identified 37 genotypes of S. mutans in these caries-active preschool children, among whom horizontal transmissions of the strains were not found.

Objective To analyze horizontal transmission patterns of Streptococcus mutans among caries-active preschool children for early interventions of dental caries. Methods Plaque samples obtained from 20 caries-active preschool children between 4 and 5 years of age were cultured under anaerobic conditions for isolating S. mutans, which were identified by morphological and biochemical analyses and PCR using primers homologous to the surface protein glucosyltransferase B (gtfB). The genotypes of the isolated S. mutans strains were determined by arbitrarily primed PCR (AP-PCR). Results Of the 200 S. mutans isolates obtained, 19 were excluded by biochemical analysis, and the remaining 181 isolates were identified as S. mutans by PCR with primers of gtfB, showing 37 different genotypes as identified by AP-PCR. Six children were found to carry S. mutans of a single genotype, 11 carried 2 genotypes, 2 had 3 genotypes, and 1 had 4 genotypes;2 children from different classes were found to carry S. mutans of the same single genotype. Conclusion We identified 37 genotypes of S. mutans in these caries-active preschool children, among whom horizontal transmissions of the strains were not found.

Objective To study the relationship between ubiquitin carboxyl terminal hydrolase L1(UCH-L1),peroxiredox protein 1(PRDX1),and the therapeutic effect of ateplase in patients with acute ischemic stroke(AIS).Methods A total of 198 AIS patients admitted to the First Hospital of Jiaxing from June 2020 to June 2022 were selected as the study group,and 100 healthy individuals who underwent physical examinations during the same period were selected as the healthy group.The serum levels of UCH-L1 and PRDX1 were detected using enzyme linked immunosorbent assay(ELISA).AIS patients were all treated with intravenous thrombolysis with ateplase.After 10 days of treatment,the clinical efficacy was evaluated.Clinical data of patients with different therapeutic effects and differences in serum UCH-L1 and PRDX1 levels were compared,and the relationship between UCH-L1,PRDX1 and the therapeutic effect of ateplase was analyzed.Results The UCH-L1 and PRDX1 of patients in the study group were higher than those in the healthy group(P<0.05).The levels of UCH-L1 and PRDX1 in severe AIS patients were higher than those in moderate and mild AIS patients(P<0.05).UCH-L1 and PRDX1 in complete anterior circulation infarction type AIS patients were higher than those in lacunar infarction,posterior circulation infarction,and partial anterior circulation infarction types(P<0.05).After treatment with ateplase,the levels of UCH-L1 and PRDX1 in AIS patients were lower than before treatment,and the worse the treatment effect of ateplase,the higher the levels of UCH-L1 and PRDX1(P<0.05).The combined prediction of UCH-L1 and PRDX1 for the efficacy of ateplase therapy in AIS patients was better than a single prediction(P<0.05).Age,severity of disease,National Institute of Health stroke scale score at admission,UCH-L1,PRDX1 are risk factors that affect the recovery of AIS patients after ateplase treatment(P<0.05).Conclusion The elevated levels of UCH-L1 and PRDX1 in the serum of AIS patients are related to the severity of the disease and stroke classification,and are risk factors affecting the efficacy of ateplase treatment in AIS patients.They can be used for early prediction of the efficacy of ateplase treatment.

OBJECTIVETo analyze horizontal transmission patterns of Streptococcus mutans among caries-active preschool children for early interventions of dental caries.

METHODSPlaque samples obtained from 20 caries-active preschool children between 4 and 5 years of age were cultured under anaerobic conditions for isolating S. mutans, which were identified by morphological and biochemical analyses and PCR using primers homologous to the surface protein glucosyltransferase B (gtfB). The genotypes of the isolated S. mutans strains were determined by arbitrarily primed PCR (AP-PCR).

RESULTSOf the 200 S. mutans isolates obtained, 19 were excluded by biochemical analysis, and the remaining 181 isolates were identified as S. mutans by PCR with primers of gtfB, showing 37 different genotypes as identified by AP-PCR. Six children were found to carry S. mutans of a single genotype, 11 carried 2 genotypes, 2 had 3 genotypes, and 1 had 4 genotypes; 2 children from different classes were found to carry S. mutans of the same single genotype.

CONCLUSIONWe identified 37 genotypes of S. mutans in these caries-active preschool children, among whom horizontal transmissions of the strains were not found.

Child, Preschool ; Dental Caries ; microbiology ; Dental Plaque ; Genotype ; Glucosyltransferases ; Humans ; Polymerase Chain Reaction ; Streptococcal Infections ; transmission ; Streptococcus mutans ; classification

OBJECTIVETo select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.

METHODSSubtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.

RESULTSDetection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.

CONCLUSIONWe have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.

Aptamers, Nucleotide ; genetics ; Cloning, Molecular ; DNA Primers ; Dental Caries ; microbiology ; Gene Library ; Humans ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Species Specificity ; Streptococcus mutans ; classification ; genetics ; isolation & purification

Objective To observe the clinical effect of traditional Chinese medicine (TCM) characteristic lung rehabilitation in treatment of patients with chronic obstructive pulmonary disease (COPD) and TCM syndrome of lung and kidney qi deficiency at stable period. Methods Sixty patients with stable COPD and lung and kidney qi deficiency syndrome admitted to the First Affiliated Hospital of Anhui University of Chinese Medicine from June to August 2017 were enrolled, and they were divided into routine treatment group and lung rehabilitation treatment group according to the random number table method, each group 30 cases. The routine treatment group was given Seretide (serevent/futicasone) dry powderi nhalation therapy; on the basis of therapy in the routine treatment group, the lung rehabilitation treatment group was treated with TCM characteristic lung rehabilitation technology (acupoint application + Chinese medicine ionic induction + oral administration of Chinese medicine Liuweibuqi granules, delivery at appropriate intervals); both groups were treated for 2 months. The changes of TCM syndrome score, western medicine symptom score, the times of acute exacerbation of COPD, COPD assessment test (CAT) score, lung function indexes: forced expiratory volume in one second (FEV1), FEV1/forced vital capacity (FVC) were observed before and after treatment in two groups. Results After treatment, TCM syndrome score, western medicine symptom score, CAT score, and after treatment the times of acute exacerbation of COPD in both groups were significantly lower than those before treatment, and the above indexes in the lung rehabilitation treatment group were markedly lower than those in routine treatment group [TCM syndrome score:11.93±1.80 vs. 14.27±2.88, western medicine symptom score: 14.20±2.75 vs. 11.93±4.23, CAT score: 14.87±2.60 vs. 16.23±4.39, the times of acute exacerbation of COPD (times): 0.63±0.49 vs. 0.95±0.83, all P < 0.05]. The improvement of FEV1 in the two groups was not significant; but FEV1/FVC in lung rehabilitation treatment group was obviously higher than that before treatment, FEV1/FVC in lung rehabilitation treatment group was significantly higher than that in the routine treatment group [(57.93±7.27)% vs. (52.49±6.61)%, P < 0.05]. Conclusion The application of TCM characteristic lung rehabilitation in the treatment of COPD patients with stable lung and kidney qi deficiency syndrome based on bronchodilators and glucocorticoids can reduce the number of acute exacerbation, improve the patients' clinical symptoms and living quality, but the improvement of lung function is not significant.

Objective:To investigate the intervention effect of topical shikonin on an imiquimod-induced psoriasis-like mouse model and its effect on expression of CCAAT enhancer binding protein δ (CEBPD) .Methods:Twenty specific pathogen-free BALB/c male mice were randomly and equally divided into model group, shikonin 1 group, shikonin 2 group and blank control group by using simple random sampling. Mice in the model group, shikonin 1 group and shikonin 2 group were topically treated with 50 mg of 5% imiquimod cream every day on the shaved back to establish the psoriasis-like mouse model. After 6-hour treatment, mice in the shikonin 1 group and shikonin 2 group were treated with 0.5 ml of shikonin at concentrations of 0.576 and 5.76 g/L respectively in the modeling area for 8 consecutive days; the blank control group received no treatment. Changes in the skin lesions of these mice were observed by naked eyes every day, and evaluated by using psoriasis area severity index (PASI) ; after 8-day treatment, the mice were sacrificed by cervical dislocation, the dorsal skin tissues were resected, and immunohistochemical study and Western blot analysis were performed to determine the expression of CEBPD in the mouse epidermis. Statistical analysis was carried out with SPSS 16.0 software by using one-way analysis of variance for comparisons of observation indices among different groups, as well as least significant difference- t test for multiple comparisons. Results:On day 8, the mice in the model group presented with obvious erythema, scales, and infiltrative and thickened skin lesions; compared with the model group, the skin lesions were markedly improved in the shikonin 1 group and shikonin 2 group, and the improvement was more obvious in the shikonin 2 group. On day 8, the PASI score significantly differed among the blank control group, model group, shikonin 1 group and shikonin 2 group (0, 11.0±1.22, 8.6±0.55, 5.8±1.30 points, respectively; F=128.21, P<0.01) , and there were significant differences between any two groups (all P < 0.01) . Immunohistochemical study showed a significant difference in the expression of CEBPD ( A value) among the model group, shikonin 1 group, shikonin 2 group and blank control group (0.072±0.026, 0.177±0.036, 0.290±0.062, 0.407±0.051, respectively; F=48.895, P < 0.01) , and there were also significant differences between any two groups (all P < 0.01) . Western blot analysis showed that the CEBPD expression in the mouse epidermis was highest in the blank control group, followed in descending order by the shikonin 2 group, shikonin 1 group and model group, and significantly differed among the above 4 groups ( F=10.237, P<0.05) ; moreover, there were significant differences in the CEBPD expression between the model group and blank control group, as well as between the shikonin 1 group and blank control group (both P<0.05) , while no significant difference was observed between the shikonin 2 group and the blank control group ( P > 0.05) . Conclusion:Topical shikonin could effectively interfere with the development of imiquimod-induced psoriasis-like mouse model; CEBPD expression decreased in the psoriasis-like mouse model, and could be markedly upregulated by topical application of shikonin.

Objective:To explore the effect of dialectical diet on traditional Chinese medicine (TCM) syndrome score of cirrhotic ascites patients based on "Gu Ben Kai Qu" theory.Methods:From March 2019 to January 2020, 84 patients with liver cirrhosis and ascites admitted to Shuguang Hospital Affiliated to Shanghai University of TCM were randomly divided into two groups according to the different dialectical types of the subjects, 14 cases in each group. Three non-syndrome differentiation diet groups were given routine nursing care of liver cirrhosis ascites. On the basis of routine nursing, the corresponding medicinal diet was selected according to syndrome differentiation based on "Gu Ben Kai Qu" theory. Patients with spleen and kidney yang deficiency syndrome selected Shenqi lean meat decoction. Patients with Yin deficiency of liver and kidney selected Wolfberry and ophiopogon spareribs decoction. Patients with qi stagnation and blood stasis syndrome selected Danggui Sanqi spareribs decoction. The TCM syndrome score scale for liver disease and the curative effect evaluation of cirrhosis ascites were used to evaluate the effect.Results:Eighty effective cases were included. On the first day of admission, the 14th day and the second week after discharge, the TCM syndrome scores of liver disease were as follows: the group (a1b1) with the spleen and kidney yang deficiency syndrome was 46.38±8.56, 34.20±8.42, 31.40±4.22, respectively. The group (a1b2) with the liver kidney yin deficiency syndrome was 41.50±8.71, 31.35±8.63, 31.12±4.94. The group(a1b3) with the qi stagnation and blood stasis syndrome was 45.92±7.86, 35.17±7.57, 30.83±7.32, respectively. The non-syndrome differentiation diet group (a2b1) with the spleen and kidney yang deficiency syndrome was 46.29±8.38, 39.79±7.65, 36.64±6.83, respectively. The non-syndrome differentiation diet group (a2b2) with the liver and kidney yin deficiency syndrome was 40.50±8.12, 38.10±8.93, 35.38±8.24, respectively. The non-syndrome differentiation diet group (a2b3) with the qi stagnation and blood stasis syndrome was 45.62±7.99, 41.83±7.31, 38.83±7.96, respectively. The comparison of TCM syndrome scores of liver disease at three time points was statistically significant ( χ2 value was 63.998, P<0.05), and the comparison between groups was statistically significant ( χ2 value was 20.993, P<0.05). On the 14th day and the second week after discharge, there were significant differences between the groups with the syndrome differentiation diet and another three groups with non-syndrome differentiation diet ( F values were 3.244, 3.489, all P<0.05). Conclusions:Based on the theory of "strengthening the foundation and opening channels", the syndrome differentiation group can effectively reduce the TCM syndrome score of patients with cirrhosis ascites, improve the symptoms and enhance the curative effect. With the development of time, the score of TCM syndrome in patients with liver disease become lower. On the 14th day of admission, patients with Yin deficiency of liver and kidney given medicated diets had significant effect; patients with spleen kidney yang deficiency syndrome or qi stagnation and blood stasis had significant effect in 2 weeks after discharge; which can effectively improve the clinical symptoms of patients with cirrhosis ascites to worthy of clinical application.

With the increasing severity of global aging, aging-related issues have become the hotspot in the field of health. In recent years, animal aging models have been widely developed and applied, which is of great significance in the study of aging mechanism. Animals with short life span, such as Caenorhabditis Elegans and Drosophila Melanogaster, have natural advantages in the study of aging. Various rat and mouse aging models have been used in aging studies. In recent years, new animal aging models have been developed, such as the African turquoise killifish. The authors reviewed main animal models used in the study of aging, and analyzed the establishment methods, evaluation indexes, advantages and disadvantages of each model in order to provide reference for related research.