Construction of EF-G knockdown strain of Mycobacterium smegmatis and drug resistance analysis / 生物工程学报

As the only translational factor that plays a critical role in two translational processes (elongation and ribosome regeneration), GTPase elongation factor G (EF-G) is a potential target for antimicrobial agents. Both Mycobacterium smegmatis and Mycobacterium tuberculosis have two EF-G homologous coding genes, MsmEFG1 (MSMEG_1400) and MsmEFG2 (MSMEG_6535), fusA1 (Rv0684) and fusA2 (Rv0120c), respectively. MsmEFG1 (MSMEG_1400) and fusA1 (Rv0684) were identified as essential genes for bacterial growth by gene mutation library and bioinformatic analysis. To investigate the biological function and characteristics of EF-G in mycobacterium, two induced EF-G knockdown strains (Msm-ΔEFG1(KD) and Msm-ΔEFG2(KD)) from Mycobacterium smegmatis were constructed by clustered regularly interspaced short palindromic repeats interference (CRISPRi) technique. EF-G2 knockdown had no effect on bacterial growth, while EF-G1 knockdown significantly retarded the growth of mycobacterium, weakened the film-forming ability, changed the colony morphology, and increased the length of mycobacterium. It was speculated that EF-G might be involved in the division of bacteria. Minimal inhibitory concentration assay showed that inhibition of EF-G1 expression enhanced the sensitivity of mycobacterium to rifampicin, isoniazid, erythromycin, fucidic acid, capreomycin and other antibacterial agents, suggesting that EF-G1 might be a potential target for screening anti-tuberculosis drugs in the future.

Protective Effect of Water Extract of Citri Grandis Exocarpium on Alcohol-induced Acute Liver Injury / 中国实验方剂学杂志

ObjectiveTo explore the pharmacodynamic effect of the water extract of Citri Grandis exocarpium （WEC） on mice with alcohol-induced acute liver injury and provide data support for the development of this medicinal for anti-alcoholism and liver protection. MethodThe main components of WEC were determined by high performance liquid chromatography （HPLC）. Sixty Balb/c mice were randomized into 6 groups： control group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， model group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， low-， medium-， and high-dose WEC groups （0.5， 1.0， 2.0 g·kg-1）， and Haiwang Jinzun tablet positive control group （2.0 g·kg-1）. The administration lasted 14 days. One day before the end of the administration， mice were fasted for 12 h with free access to water. The mice， except the control group， were given 56° Chinese liquor （13 mL·kg-1）. After 2 h， blood was taken from eyeballs and the liver was dissected and weighed. Automatic biochemical analyzer was employed to detect the expression of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and alcohol dehydrogenase （ADH）. The pathological changes of liver tissues were observed based on hematoxylin-eosin （HE） staining， and apoptosis of hepatocytes based on TUNEL/DAB staining. The expression of proteins related to apoptosis was detected by Western blot. ResultAccording to the HPLC fingerprint， the main components of WEC were rhoifolin and naringin. Compared with the control group， the model group showed increase in liver/body weight ratio （P<0.01） and the expression of ALT and AST （P<0.05， P<0.01）， decrease in the expression of ADH （P<0.05）， blurred structure of hepatic lobules， pathological changes of liver tissue， loose cytoplasm with edema， severe steatosis， rise of the TUNEL-positive rate （P<0.01）， reduction in expression of Bcl-2 （P<0.01）， and increase in Bax and Caspase-3 （P<0.01）. Compared with the model group， medium-dose WEC lowered liver/body weight ratio （P<0.05）. All doses of WEC depressed the activity of ALT and AST （P<0.05， P<0.01）， up-regulated the expression of ADH （P<0.05）， significantly improved the pathological features of alcohol-induced cytoplasmic porosity， edema， and steatosis， down-regulated the TUNEL-positive rate （P<0.05， P<0.01）， enhanced the expression of Bcl-2 （P<0.05）， and decreased Bax and Caspase-3 （P<0.01）. ConclusionWEC regulates the expression of ALT， AST， and ADH and improves hepatic steatosis and hepatocyte apoptosis to fight against acute liver injury.

Protective Effect of Water Extract of Citri Grandis Exocarpium on Alcohol-induced Acute Liver Injury / 中国实验方剂学杂志

ObjectiveTo explore the pharmacodynamic effect of the water extract of Citri Grandis exocarpium （WEC） on mice with alcohol-induced acute liver injury and provide data support for the development of this medicinal for anti-alcoholism and liver protection. MethodThe main components of WEC were determined by high performance liquid chromatography （HPLC）. Sixty Balb/c mice were randomized into 6 groups： control group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， model group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， low-， medium-， and high-dose WEC groups （0.5， 1.0， 2.0 g·kg-1）， and Haiwang Jinzun tablet positive control group （2.0 g·kg-1）. The administration lasted 14 days. One day before the end of the administration， mice were fasted for 12 h with free access to water. The mice， except the control group， were given 56° Chinese liquor （13 mL·kg-1）. After 2 h， blood was taken from eyeballs and the liver was dissected and weighed. Automatic biochemical analyzer was employed to detect the expression of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and alcohol dehydrogenase （ADH）. The pathological changes of liver tissues were observed based on hematoxylin-eosin （HE） staining， and apoptosis of hepatocytes based on TUNEL/DAB staining. The expression of proteins related to apoptosis was detected by Western blot. ResultAccording to the HPLC fingerprint， the main components of WEC were rhoifolin and naringin. Compared with the control group， the model group showed increase in liver/body weight ratio （P<0.01） and the expression of ALT and AST （P<0.05， P<0.01）， decrease in the expression of ADH （P<0.05）， blurred structure of hepatic lobules， pathological changes of liver tissue， loose cytoplasm with edema， severe steatosis， rise of the TUNEL-positive rate （P<0.01）， reduction in expression of Bcl-2 （P<0.01）， and increase in Bax and Caspase-3 （P<0.01）. Compared with the model group， medium-dose WEC lowered liver/body weight ratio （P<0.05）. All doses of WEC depressed the activity of ALT and AST （P<0.05， P<0.01）， up-regulated the expression of ADH （P<0.05）， significantly improved the pathological features of alcohol-induced cytoplasmic porosity， edema， and steatosis， down-regulated the TUNEL-positive rate （P<0.05， P<0.01）， enhanced the expression of Bcl-2 （P<0.05）， and decreased Bax and Caspase-3 （P<0.01）. ConclusionWEC regulates the expression of ALT， AST， and ADH and improves hepatic steatosis and hepatocyte apoptosis to fight against acute liver injury.

Effects of Crataegolic Acid on Inflammatory Response and Oxidative Stress in Non-alcoholic Fatty Liver Disease Model Mice Induced by High-fat Diet / 中国药房

OBJECTIVE： To investigate the effects of crataegolic acid on inflammatory response and oxidative stress in non-alcoholic fatty liver disease （NAFLD） model mice induced by high-fat diet. METHODS： Totally 72 C57BL/6 mice were randomly divided into normal group （normal saline）， model group （normal saline）， simvastatin group （positive control， 3 mg/kg） and crataegolic acid low-dose， medium-dose and high-dose groups （25， 50 and 100 mg/kg）， with 12 mice in each group. Normal group was standard feed， while other groups were given high-fat diet to induce NAFLD model . At the same time of modeling， rats in each group was given relevant dose of drugs intragastrically， once a day， for consecutive 12 weeks. 12 h after last administration， body weight and liver weight of mice were determined， and liver index was calculated. The changes of serum biochemical indexes （activities of AST， ALT and contents of HDL-C， LDL-C， TG， TC） were determined by fully automatic biochemical analyzer. The pathological changes of liver tissue were observed after HE staining. The changes of inflammatory response indexes （contents of NF-κB， TNF-α， IL-6） in liver tissue were determined by ELISA， and the changes of oxidant stress indexes （contents of MDA， SOD and activity of GSH-Px） were determined by spectrophotometry. RESULTS： Compared with normal group， the liver index of mice in model group was increased significantly （P＜0.05）. The serum content of HDL-C was decreased significantly， and the activities/contents of other serum biochemical indexes were increased significantly （P＜0.05）. The boundary of hepatic lobules was not clear， and the liver tissue had obvious pathological changes. Inflammatory response indexes and the contents of MDA were increased significantly in liver tissue （P＜0.05）， the activities of SOD and GSH-Px were decreased significantly （P＜0.05）. Compared with model group， except that the increase of SOD activity and the decrease of MDA content of liver tissue were not significantly in crataegolic acid low-dose group （P＞0.05）， while above indexes of mice in other administration groups were improved significantly （P＜0.05）. The globular lipid droplets and inflammatory cell infiltration were decreased in liver tissue. CONCLUSIONS： Crataegolic acid can effectively alleviate the degree of liver lesions in NAFLD model mice induced by high-fat die through anti-inflammatory and inhibiting oxidative stress.

Whole exome sequencing identifies an AMBN missense mutation causing severe autosomal-dominant amelogenesis imperfecta and dentin disorders / 国际口腔科学杂志·英文版

Tooth development is a complex process that involves precise and time-dependent orchestration of multiple genetic, molecular, and cellular interactions. Ameloblastin (AMBN, also named "amelin" or "sheathlin") is the second most abundant enamel matrix protein known to have a key role in amelogenesis. Amelogenesis imperfecta (AI [MIM: 104500]) refers to a genetically and phenotypically heterogeneous group of conditions characterized by inherited developmental enamel defects. The hereditary dentin disorders comprise a variety of autosomal-dominant genetic symptoms characterized by abnormal dentin structure affecting either the primary or both the primary and secondary teeth. The vital role of Ambn in amelogenesis has been confirmed experimentally using mouse models. Only two cases have been reported of mutations of AMBN associated with non-syndromic human AI. However, no AMBN missense mutations have been reported to be associated with both human AI and dentin disorders. We recruited one kindred with autosomal-dominant amelogenesis imperfecta (ADAI) and dentinogenesis imperfecta/dysplasia characterized by generalized severe enamel and dentin defects. Whole exome sequencing of the proband identified a novel heterozygous C-T point mutation at nucleotide position 1069 of the AMBN gene, causing a Pro to Ser mutation at the conserved amino acid position 357 of the protein. Exfoliated third molar teeth from the affected family members were found to have enamel and dentin of lower mineral density than control teeth, with thinner and easily fractured enamel, short and thick roots, and pulp obliteration. This study demonstrates, for the first time, that an AMBN missense mutation causes non-syndromic human AI and dentin disorders.

Effect of Ronghuang Granule on serum FGF23, FGFRs and Klotho in non-dialysis patients with CKD-MBD and kidney deficiency and damp-heat syndrome / 南方医科大学学报

OBJECTIVE@#To observe the effect of Ronghuang granule on serum fibroblast growth factor 23 (FGF23), fibroblast growth factor receptor (FGFRs) and Klotho protein levels in non-dialysis patients with chronic kidney disease-mineral and bone disorder (CKD-MBD) and kidney deficiency and damp heat syndrome.@*METHODS@#Seventy non-dialysis CKD-MBD patients with kidney deficiency and dampness-heat syndrome were randomized into control group (=35) and treatment group (=35). All the patients were given routine treatment combined with traditional Chinese medicine retention enema, and the patients in the treatment group received additional Ronghuang granule treatment (3 times a day). After the 12-week treatments, the patients were examined for changes of TCM syndromes. Serum levels of Ca, P, parathyroid hormone (iPTH), FGF23, FGFRs and Klotho proteins were detected before and after treatment. These parameters were also examined in 20 healthy volunteers.@*RESULTS@#Sixty-five patients completed the study, including 33 in the control group and 32 in the treatment group. The patients in the treatment group showed significantly better treatment responses than those in the control group ( < 0.05 or 0.01). At 4, 8, and 12 weeks of treatment, the patients in the treatment group had significantly lowered scores of TCM syndromes compared with the score before treatment ( < 0.05 or 0.01), while in the control group, significant reduction of the scores occurred only at 12 weeks ( < 0.05); at each of the time points, the treatment group had significantly greater reductions in the score than the control group ( < 0.01). Significant improvements in serum Ca, P and iPTH levels were observed at 4, 8, and 12 weeks in the treatment group ( < 0.05) but only at 12 weeks in the control group ( < 0.05). The patients in the control and treatment groups all showed elevated serum levels of FGF23, FGFRs and Klotho protein compared with the normal subjects ( < 0.01); FGF23, FGFRs and Klotho levels were significantly reduced in the treatment group ( < 0.05) but remained unchanged in the control group (>0.05), showing significant differences between the two groups.@*CONCLUSIONS@#Ronghuang granule improves the clinical symptoms of non-dialysis CKD-MBD patients with kidney deficiency and dampness heat syndrome by reducing serum levels of FGF23, FGFRs and Klotho, improving calcium and phosphorus metabolism disorder, and inhibiting secondary hyperparathyroidism.

Analysis of distribution characteristics of clinical isolates of nontuberculous mycobacteria in a general hospital / 中华传染病杂志

Objective To investigate the current prevalence of nontuberculous mycobacteria (NTM) in Shanghai,and to study the distribution characteristics of NTM clinical isolates,which may help to improve the diagnostic level of NTM and provide guidance for effective prevention and treatment of NTM infection.Methods Culture-positive isolates of clinical mycobacteria were collected from 2008 to 2013 in Huashan Hospital affiliated to Fudan University.All isolates were heat inactivated,and the genomic DNA was extracted and the species were identified by comprehensive comparative analysis of 16S rDNA,hsp65 and rpoB target genes sequencing.Results From January 2008 to December 2013,the overall mycobacterial culture-positive rate was 4.1 ％ (411/10 015).After excluding the repeated isolates,a total of 253 culture-positive mycobacteria isolates were collected for the species identification.By genes sequencing analysis,140 isolates were identified as mycobacterium tuberculosis complex (MTBc),102 NTM and 11 Nocardias,accounting for 55.3％,40.3％ and 4.4％,respectively.Positive rate of NTM isolates had an increasing trend from 25.0％ in 2008 to 42.7％ in 2013,reaching a highest rate of 54.9％ in 2012.In further analysis of 102 NTM isolates,16 species were identified.Among them,28 were M.abscesses,18 strains of M.marinum,17 strains of M.avium-intracellulare complex and 10 strains of M.fortuitum,accounted for 27.5％,17.6％,16.7％ and 9.8％,respectively.Conclusions Both of the isolation number and isolation rate of NTM in the general hospital are increasing.NTM related cases are also increasing in recent years,which mainly caused by M.abscess,M.marinum,M.aviumintracellulare complex and M.fortuitum.

Pathogen isolation and whole-genome sequence analysis of human adenovirus type 55 outbreak on plateaus / 军事医学

Objective To investigate the variation characteristics of adenovirus type 55 (HAdV-B55) gene on plateaus.Methods Throat swabs were collected from HAdV-B55 infected patients and used for virus isolation in HEp-2 cells.The whole-genome sequence was obtained by PCR and sequencing.HAdV-B55 gene sequence was blast with the previously reported virus.Results HAdV-B55 strains were isolated from throat swabs, which were named LS89/Tibet/2016.The whole-genome sequence was obtained and submitted to GenBank with the accession number of KY002683.No large fragment gene recombination was found between this HAdV-B55 strain and previous strains, and the sequence similarity with QS-DLL strain was 99.9%.Conclusion This study provides more information for the evolution patterns of adenovirus 55 and will contribute to the prevention and control of HAdV-B55 infection in the future.

Correlation between optical density and colony forming units of Mycobac-terium tuberculosis suspension / 中国感染控制杂志

Objective To establish a reliable approach for quantification of colony forming unit(CFU)of Mycobac-terium tuberculosis (M.tb)by measuring optical density(OD).Methods M.tb suspension H37Ra was prepared using low-power ultrasonic or glass bead beating methods,and was two-fold serially diluted,OD at 600nm (OD600)of each dilution ratio was measured respectively,OD600 and dilution curve were analyzed to determine the optimum approach for preparing bacterial suspension,linear range of OD600,as well as linear relationship between OD600 and CFU.Results OD600 was 0.1 -0.6,linear regression analysis of OD600 and dilution ratio within linear range revealed that correlation coefficient (R2 )of glass bead beating and low-power ultrasonic methods were 0.98 and 1 .00 respectively,both presented a good correlation,low-power ultrasonic method was better than glass bead beat-ing method,bacterial suspension dispersed more evenly.Linear regression analysis results of OD600 and CFU val-ues showed that the regression equation of glass bead beating method and low-power ultrasonic method were CFU=2.35×107 ×OD600+4.42×105 and CFU=3.26×107 ×OD600+6.89×105 respectively.Conclusion Low-power ultrasonic method is a good method for preparation of M.tb suspension,combined the measurement of OD600 value, it can be a reliable and rapid method for quantitative analysis of M.tb.

Impact of subclinical hypothyroidism on macro-vascular complications in elderly type 2 diabetic patients / 中华老年医学杂志

Objective To investigate the impact of subclinical hypothyroidism (SCH) on macrovascular complications in elderly type 2 diabetic patients.Methods A total of 1170 hospitalized elderly patients with type 2 diabetes mellitus were enrolled in the study through systematic sampling and underwent testing for blood biochemical indicators, thyroid function and C peptide.Parameters for macro-vascular complications, including the ankle/brachial index (ABI), transcranial Doppler vascular ultrasound (TCD), electrocardiogram (ECG), ejection fraction (EF), history of coronary heart disease, and hypertension grading were also monitored.Results All the subjects were divided into two groups based on the thyroid stimulating hormone (TSH) level: the euthyroid group (4 mU/L≥TSH＞0.4 mU/L) and the SCH group (TSH＞4 mU/L), and the latter was further sub-grouped into the mild SCH group (10 mU/L≥TSH＞4 mU/L) and the severe SCH group (TSH＞10 mU/L).ABI was significantly decreased in SCH (R/L: 0.86/0.92, P＜0.01).Levels of basal C-peptide (CP0) and post glucose-challenge C-peptide (CP1-3) were higher in the SCH group than in the euthyroid group [(2.16±0.93)pg/L vs.(1.56±1.05)pg/L, (0.53±0.25)pg/L v, (0.38±0.37),(0.72±0.23) pg/L vs.(0.56 ±0.32) pg/L, (6.21± 2.69) pg/L vs.(4.46 ± 2.62) pg/L,respectively, P＜0.01 for all].EF was higher in the SCH group than in the euthyroid group[(70.87± 6.66)％ vs.(65.10 ± 8.08％), P＜ 0.01].There were no significant differences in other biochemical indicators, ECG, TCD, history of coronary heart disease, hypertension grading and intervention treatment (P＞0.05 for all).Conclusions Lower extremity atherosclerotic disease has a higher incidence in elderly type 2 diabetic patients with SCH and occurs earlier than other macrovascular complications.Elevated TSH levels and insulin resistance may be the major causes.

A study on Qingshen granule for treatment of renal fibrosis in patients with chronic renal failure accompanied by damp-heat syndrome and its mechanism / 中国中西医结合急救杂志

Objective To observe the anti-fibrosis therapeutic effect and mechanism of Qingshen granule for treatment of patients with chronic renal failure (CRF) accompanied by damp-heat syndrome.Methods Sixty-eight patients with CRF accompanied by damp-heat syndrome were randomly divided into a control group and a observation group, and the study was completed only in 61 patients, 31 in the control group and 30 in the observation group. Thirty subjects having taken physical health examination were assigned in a healthy control group. All the patients in both treatment groups were treated with conventional western medical therapy and traditional Chinese medicine (TCM) retention enema, and for patients in observation group, Qingshen granule was given additionally, 1 bag (10 g) thrice a day taken orally. The therapeutic course was 8 weeks. The clinical therapeutic effect, the levels of serum creatinine (SCr), the glomerular filtration rate (eGFR), serum interleukin-17 (IL-17), collagen type Ⅲ (Col-Ⅲ) and nuclear factor-κB p65 (NF-κB p65) in peripheral blood mononuclear cells (PBMC) were measured before and after treatment in the two treatment groups, and the above results were compared with those in healthy control group.Results Clinically, the total effective rates of the disease and of the TCM syndrome in observation group were significantly higher than those in the control group (86.67% vs. 58.06%, 83.33% vs. 45.16%, bothP < 0.01). In the observation group, the level of SCr was obviously lower, and the level of eGFR was markedly higher after treatment, and compared with the control group, the changes in above data after treatment in observation group were more significant [SCr (μmol/L): 250.62±164.97 vs. 393.72±183.64, eGFR (mL·min-1·1.73 m-2): 33.42±17.24 vs. 39.72±23.85, bothP < 0.05]. After treatment, the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC were obviously lowered in both treatment groups compared with those before treatment, the therapeutic effect in observation group being superior to that in the control group [IL-17 (ng/L): 17.47±8.87 vs. 25.51±16.69, Col-Ⅲ (μg/L): 17.06±8.76 vs. 23.77±10.44, NF-κB p65 (μg/L): 0.58±0.34 vs. 0.83±0.30, allP < 0.05].Conclusion The Qingshen granule can ameliorate the clinical symptoms, improve renal function, decrease the levels of serum IL-17, Col-Ⅲ and NF-κB p65 in PBMC, intervene renal fibrosis in patients with CRF and damp-heat syndrome, ultimately delaying the progress of CRF.

The Experimental Study on Dual Antiplatelet Drugs-Induced Intestinal Damage in Rats / 天津医药

Objective To investigate the effect of dual antiplatelet drugs on the intestinal damage in rats. Meth-ods Eighty SD rats were randomly allocated into four groups:control group (normal saline, n=20), aspirin group (10.41 mg/kg, n=20), clopidogrel group (7.81 mg/kg, n=20) and clopidogrel combined aspirin group (n=20). Each group was given intra-gastric administration of drugs once per day for 14 days. All rats received operation after the final intragastric administration. The intestinal injury was observed. The jejunal fluid was taken for bacterial culture. The intestinal permeability was detected by ethidium bromide (EB) method. The small intestinal mucosal injury was estimated by Chiu method. Results The differ-ent degrees of small intestinal mucosal injury were found in four groups. The scores of pathological lesions were significantly higher in aspirin group, clopidogrel group and clopidogrel combined aspirin group than those of control group (P<0.05). The dual antiplatelet group showed the highest score (3.450±1.356). The small intestinal permeability was significantly increased in experiment groups compared with that of control group (54.012±3.513μg/g, P<0.05). There was a significantly higher small intestinal permeability in dual antiplatelet group than that of aspirin group and clopidogrel group (μg/g:130.533 ± 29.631 vs 90.965±3.765 vs 66.800±4.853, P<0.001). The total jejunal bacteria was significantly increased in dual antiplate-let group than that of control group (CFU/mL:61924.805±1751.159 vs 18154.280±1153.376, P<0.001). The ratio of entero-bacterium and enterococcus was significantly decreased in dual antiplatelet group compared with that of control group (0.220±0.089 vs 1.007±0.148, P<0.001). Conclusion The routine dose of dual antiplatelet drug aggravates the small intes-tinal injury in rats compared with that of single drug. The manifestations of intestinal mucosal injury include increased intes-tinal bacteria, dysbacteriosis, and increased small intestinal permeability.

The relationship between subclinical hypothyroidism and diabetic retinopathy in elderly type 2 diabetic patients / 中华老年医学杂志

Objective To investigate the relationship between subclinical hypothyroidism (SCH) and diabetic retinopathy (DR) in elderly type 2 diabetic patients.Methods 1170 hospitalized patients with type 2 diabetes mellitus were enrolled in our study.After identified by systematic sampling,all the elderly patients received examinations of thyroid function,biochemical indexes,steamed bread meal test and C-peptide releasing test.The serious degree of DR was observed through digital fundus photography and ophthalmofundoscope.The relationship between SCH and DR was analyzed.Results All the subjects were divided into two subgroups:euthyroid group(TSH≤4mU/L)and subclinical hypothyroidism (SCH) group (mild SCH group,TSH ＞4～10 mU/L; serious SCH group,TSH＞10 mU/L).Free thyroxine (FT4) was significantly lower in SCH group than in euthyroid group[(13.91±2.17) pmol/L vs.(16.55±2.81)pmol/L,P＜0.001].The levels of basal (CP0) and post glucose-challenge (CP1-3) C-peptide were significantly higher in SCH group than in euthyroid group (P0-3 value:0.001,0.012,0.004,0.001).There were significant differences in the progress of diabetic retinopathy between SCH and euthyroid groups (retinopathy-L,P=0.018; retinopathy-R P=0.013).Conclusions Elderly type 2 diabetic patients with SCH have a higher incidence of diabetic retinopathy (DR).The elevated FT4 level and decreased basal C-peptide level are probably the main reasons.

Effects of hypoxia on circulating tumor cells in gastric adenocarcinoma in vivo / 中华消化杂志

Objective To explore the correlation between the change of hypoxic in microenvironment and the genesis of circulating tumor cells (CTCs) in gastric adenocarcinoma.Methods After human gastric adenocarcinomn BGC-823 cells treated with different concentration of CoCl2 (0,100,200,300,400 μmol/L)for four hours,the expression levels of hypoxia inducible factor 1 alpha (HIF-1α)were detected by Western blot to select the optimal concentration.A total of 40 BALB/c mice were divided into two equal groups,named hypoxic group and normoxic group.Optimal concentration of CoCl2 treated BGC-823 gastric cancer cells and non hypoxic induced BGC 823 gastric cancer cells were injected through the tail vein.After three days blood samples were collected through the angular vein.CTCs were enriched by density gradient centrifugation,identified with cytokeratin19 (CK19) antibody marker by cell immunofluorescence and cell number was counted.Statistical significace test were performed by analysis of variance and Dunnett-T test.Results The expression of HIF-1α reached a maximun of 1.40±-0.16 when BGC-823 cells were treated with 200 μmol/L CoCl2 for four hours.The number of CTCs in hypoxic group was higher than that of normoxic group ((4.70± 1.72)/high power field (HPF) vs (1.15±0.56)/HPF,Z=-4.824,P＜0.01).Conclusion Hypoxic microenvironment might promote the genesis of CTC in gastric adenocarcinoma.

Analysis of 5 Dyes Residues in Navel Orange with Temperature-assisted Ionic Liquid Dispersive Liquid-liquid Microextraction and High Performance Liquid Chromatography Detection / 分析化学

A fast method composed of the quick, easy, cheap, effective, rugged and safe ( QuEChERS) and temperature-assisted ionic liquid dispersive liquid-liquid microextraction ( TA-IL-DLLME) sample preparation coupled with high performance liquid chromatography ( HPLC ) for the analysis of 5 dyes residues in navel orange was developed. The QuEChERS sample preparation involved the quick extraction with acetonitrile in the presence of anhydrous MgSO4 and NaCl and the purification with primary secondary amine ( PSA ) sorbent. The TA-IL-DLLME sample preparation was processed using 1 mL of the extract obtained by QuEChERS as dispersive solvent and 60 μL of 1-octyl-3-methylimidazolium hexafluorophosphate as extractive solvent under 55 ℃ of water-bath temperature and 12 min of water-bath time. The ultimate solution was detected by HPLC-UV and the contaminated sample was further confirmed by UPLC-MS/MS under multiple reactions monitoring (MRM) mode. The recoveries of five dyes were in the range from 70. 3% to 93. 6% at two spike levels of 0. 01 and 0. 05 mg/kg, the relative standard deviations (RSDs) were between 3. 5% and 9. 2% and the limits of quantification (LOQs) were between 1. 1 and 2. 8 μg/kg.

Transplantation of adipose-derived stem cells with fibrin glue for treatment of acute myocardial infarction in rat / 中华急诊医学杂志

Objective To investigate the cell survival of the combination of fibrin glue and adiposederived stem cells (ADSCs) in rats when implanted into ischemic myocardium and the improvement of heart function.Methods The rat ADSCs were isolated from the subcutaneous adipose tissues.The surface phenotype of these cells was analyzed by flow cytometry.Myocardial infarction was induced in female rats using coronary artery ligation.One week after MI,surviving rats were randomized (random nuber) into 4 groups,control group (n =10),fibrin group (n =10),cell group (n =10) and combination group (n =10).100 μl of PBS was injected into the ischemic myocardium in control group.100 μl of Fibrin glue were injected into ischemic myocardium in fibrin group.100 μl of ADSCs labeled with DAPI were injected into the infract along the border zone in cell group.ADSCs in 100 μl of fibrin glue were injected into the infract in combination group.Four weeks after the injection the surviving rats underwent examination of heart functions by the Hemodynamics.The rats were killed and their hearts were taken out to undergo immunohistochemistry with 4,6-diamidino-2-phenylindole (DAPI) and actin and factor Ⅶ to measure the area of cardiac infarction and the capillary density.The heart infarcted size was calculated by masson trichrome staining.All data was analyzed by software SPSS 15.0,ANOVA comparison tests and the student t test were used,and P ＜ 0.05 was considered as statistically significant.Results Four weeks after the cells were transplanted,LVSP and + dp/dtmax of combination group were highest among all groups.The heart infarcted size of the combination group was (28.5 ± 3.6) ％,significantly less than those of the cell group (33.33 ± 2.3) ％ and fibrin group (35.96 ± 2.11) ％,both P ＜ 0.05.The capillary density of the combination group was (108.7 ± 11.38) /mm2,significantly greater than those of the cell group and that of the fibrin group,and greater than that of the control group.DAPI and actin double staining detected a varied increase in the number of surviving cardiomyoctyes at the heart infarcted area.Conclusions Transplantation of ADSCs with fibrin glue brings better improvement in cell survival and in restoration of heart function than either cellular or fibrin therapy alone.

The diagnostic value of multi-slice CT on the congenital malformation of coronary sinus / 中华放射学杂志

Objective To investigate the application value of multi-slice spiral CT on the congenital malformation of coronary sinus. Methods MSCT finding of 98 patients with coronary sinus malformation confirmed by surgery were retrospectively analyzed,and the cases were divided into four categories based on the Mantini theory and comparison was made between the diagnosis from ultrasound and CT.A 2 × 2 table for Chi-square test was also used for statistics analysis.Results Among 98 patients,there were 72 patients with persistent left superior vena cava reflowed to right atria through coronary sinus,with 48 patients diagnosed by ultrasound and 72 patients by MSCT; there were 13 patients with anomalous pulmonary venous connection to coronary sinus,with 12 patients diagnosed by ultrasound and 13 patients by MSCT diagnosis; there were 10 patients with unroofed coronary sinus syndrome,with 6 patients diagnosed by ultrasound and 8 patients by MSCT,there were 2 patients with coronary sinus atresia,all diagnosed by MSCT; there were 1 patient with coronary sinus anomaly reflow to left arita.The significant difference between 2 modalities (x2 =22.7,P＜0.01) shows that CT is superior to ultrasound.Conclusion MSCT is much more better than ultrasound in the diagnosis of malformation of coronary sinus and it can provide reliable diagnosis prior to surgery or interventional therapy.

Comparison of measurements for ALT and AST among four domestic reference laboratories / 中华检验医学杂志

Objective To investigate the intralaboratury and interlabomtory variations of measurements for ALT and AST among four domestic reference laboratories. Methods The International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) reference procedures and IFCC procedures without pyridoxal 5-phosphate (PLP) were performed in the reference laboratories. Intralaboratory and interlaboratory CVs were compared with those in 2006 and 2007 IFCC External Quality Assessment Scheme for Reference Laboratories (RELA). Meanwhile, deviations of results for ALT, AST and AST/ALT between two methods were calculated. Results Interlaboratory CVs were generally higher than intralaboratory CVs. Interlaboratory CVs among the 41 laboratories were lower than these in RELA. Results of ALT and AST using method with PLP were higher than those using method without PLP. Difference of AST/ALT ratio between the two methods was significant. Conclusions For reference measurement of the 2 enzymes, interlaboratory CVs of < 3.5 are achievable on frozen serum materials. Measurements on lyophilized materials may have higher CVs. Further studies are needed for the investigation of the differences between results obtained in the absence and presence of PLP.

Efficacy and safety of glargine combined with repaglinide in treatment of type 2 diabetes mellitus in the elderly / 中华老年医学杂志

Objective To compare the blood glucose level and associated hypoglycemia risks between group of insulin glargine combined with repaglinide and group of mixed insulin in the treatment of type 2 diabetes in the elderly. Methods Sixty four type 2 diabetes patients (age≥60 years) with inadequate glyeaemic control by drugs were divided into 2 groups randomly: glargine group (32 cases) and mixed insulin group (32 cases). In glargine group, 32 patients were given repaglinide before breakfast, lunch, supper respectively and injection of glargine hypodermically once at 22 o'clock every night, while the 32 patients in mixed insulin group were injected with the mixed insulin (Novolin 30R) hypodermically before breakfast and supper respectively for 16 weeks. The dose of repaglinide and insulin were adjusted every 3 days according to the level of fasting glucose (FPG)and postprandial glucose (PPG), reaching the aim of FPG less than 7.2 mmol/L and PPG less than 10mmol/L. The blood glucose level and the incidence of hypoglycemia were observed. Results The daily glucose profile and the level of HbAlc of the 2 groups dropped obviously after 16 weeks treatments (all P<0. 05). At the end of the experiment, the PPG of lunch and supper, and the level of HbAlc were markedly lower in glargine group than in mixed insulin group (all P<0. 05), and the body mass index (BMI) increased markedly in mixed insulin group compared with pre-experiment (P<0. 05), but no significant change was found in glargine group (P>0. 05). The incidence of hypoglycemia in glargine group was lower than that in mixed insulin group (2 patients in glargine group, 12 patients in mixed insulin group, P<0.05). Conclusions Both of the mixed insulin and glargine combined with repaglinide have visible effects on controlling the blood glucose, but the latter has better efficacy, lower risk of hypoglycemia and lower increase of BMI than the former.

Expression of angiotensin-converting enzyme 2 in rat pancreas / 基础医学与临床

Objective To investigate the expression of angiotensin-converting enzyme 2(ACE2) in rat endocrine and exocrine tissue of pancreas and to compare its expression level with that in other visceral tissues. Methods The expression of ACE2 in rat visceral organs was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Results ACE2 was positively expressed in all the test tissues, including pancreas, heart, liver, kidney, lung, spleen, duodenum, ileum and bladder. The expression of ACE2 mRNA was detected in whole pancreas and ACE2 protein was found in both endocrine tissue and exocrine tissue of rat pancreas, while in the endocrine tissue much less ACE2 protein was expressed. The absorbance value of exocrine tissue of rat pancreas increased significantly as compared to that in pancreatic islet. Conclusion Our data demonstrated a tissue specific pattern of ACE2 expression in rat pancreas and this indicated ACE2 might be present in islet of pancreas of human, which might cause the elevation of blood glucose levels in SARS patients. Since ACE2 works as a negative regulator of the angiotensin system, the expression of ACE2 protein in endocrine tissue of pancreas may facilitate the treatment of diabetes.