Objective To detect the levels of five trace elements in whole blood of patients with Keshan disease(KSD) and dilated cardiomyopathy(DCM) and explore their role in the pathogenesis of KSD.Methods One hundred and four patients with chronic KSD were selected from Keshan diseased areas in Shandong,Sichuan and Inner Mongolia.Thirty patients with DCM were selected from Qilu Hospital of Shandong University,Jinan Central Hospital,The First People's Hospital.Ninety-one healthy people from KSD endemic areas and 39 healthy people from Jinan were selected as endemic healthy controls and non-endemic healthy controls,respectively.Blood samples were collected to determinate the level of selenium (Se),copper (Cu),zinc (Zn),chromium (Cr) and manganese (Mn) with fluorescence method and atomic absorption spectrometry,according to the principle of informed consent.Results The level of Se,Zn and Cr of KSD group[(36.0 + 4.9)μg/L,(22.73 + 4.62)mg/L,(0.56 + 0.17)mg/L] was significantly lower than that of non-endemic healthy controls [(56.4 ± 6.8)lμg/L,(25.35 ± 4.44)mg/L,(0.71 ± 0.17)mg/L,all P ＜ 0.05],but the level of Cu of KSD group[(0.95 ± 0.24)mg/L] was significantly higher than that of non-endemic healthy controls[(0.73 ± 0.13) mg/L,all P ＜ 0.05].The level of Se and Cr of KSD was significantly lower than that of endemic healthy controls[(54.5 ± 5.4)μg/L,(0.87 ± 0.02)mg/L,P ＜ 0.05],and Cu was significantly higher than that of endemic healthy controls[(0.66 ± 0.02)mg/L,P ＜ 0.05].The level of Cu and Zn of KSD was significantly lower than that of DCM [(1.21 ± 0.23)mg/L,(27.09 ± 7.10)mg/L,all P ＜ 0.01].The level of Se and Cr of DCM group[(39.6 ± 3.5)μg/L,(0.58 ± 0.14)mg/L] was significantly lower than that of non-endemic healthy controls(all P ＜ 0.01),but Cu[(1.21 + 0.23)mg/L] was significantly increased (P ＜ 0.01).Compared with non-endemic healthy controls,the level of Se of endemic healthy control group was significantly decreased (P ＜ 0.01),while Cu was significantly increased (P ＜ 0.01).Se,Zn and Cr level of KSD decreased gradually following elevated heart function level,but the level of Cu gradually increased.Conclusions The metabolism of Se,Cr,Cu and Zn is unbalanced in KSD patients,whose Se level is still lower than that of people in non-endemic areas.The change of Se,Cr,Cu and Mn level between KSD and DCM is consistent.

Objective To evaluate the clinical value of color doppler flow image (CDFI) and three-dimension color power angiography(3D-CPA) in the diagnosis of thyroid nodules. Methods A total of 62 pathologically confirmed thyroid nodules of 30 patients were analyzed. All the patients underwent ultrasound examination and operation in the Second Affiliated Hospital of Harbin Medical University between January, 2008 and May, 2009. Both CDFI and 3D-CPA were performed before the operation. All the nodules were divided into three groups including nodular goiters, thyroid adenomas and thyroid cancers according to the pathological results. The hemodynamic features and the vascular morphology characteristics of nodules in different groups were compared. Results 3D-CPA showed that blood vessels of nodular goiters commonly distributed surrounding the mass and the inner vessels were thin and regular, thyroid adenomas were bulb-shaped and netted structure, and malignant thyroid nodules displayed distorted and irregular distributed vessels. Peak systolic velocity (PSV)of the three groups were (39.43±11.17a), (46.39±12.98) and (65.17±9.23)cm/s, respectively. Resistance index(RI) of the three groups were (0.32±0.08), (0.41±0.06) and (0.69±0.07)cm/s, respectively. Both PSV and RI in malignant thyroid nodules were higher than in nodular goiters and in thyroid adenomas and the difference were statistical significant (all P < 0.05). The blood flow grade of malignant nodules was also higher in malignant nodules than in other two groups(χ2 = 17.11, 12.79, 23.05, 15.41, P< 0.01). Conclusions CDFI and 3D-CPA could visually demonstrate the characteristic and distribution of the inner and outer blood vessels, display the vessels structures, and they are benefit the differential diagnosis of thyroid nodules.

OBJECTIVETo observe the effect of calorie restriction on the high fat diet rats mRNA expressions of liver forkhead box O1(FoxO1), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G-6-P) and to explore the possible mechanisms.

METHODS24 normal 6-week-old male Wistar rats were randomly divided into three groups: normal chow group (NC, n = 7), high fat diet group (HF, n = 9) and calorie restriction group (CR, n = 8). They were fed for 12 weeks. At the end of the experiment, the rats were sacrificed and their fasting blood glucose (FBG), insulin (INS), triglycerides (TG), total cholesterol (TC) were measured. Their visceral fat (VF) and body weight (BW) were also measured and VF/BW was calculated. Gene expression was investigated by using semi-quantitative RT-PCR methods. Liver histology was studied with HE stained slides.

RESULTSCompared with the NC group, HF group rats developed visceral obesity which was accompanied by higher FBG, plasma INS, TG, and TC. The levels of FoxO1, PEPCK, and G-6-P increased by 18.9%, 33.8%, and 24.6%, respectively (P less than 0.01). Liver steatosis was observed with microscopy. The BW, VF FBG, INS, TG and TC of the CR group rats were lower in comparison to those of the HF group. The levels of FoxO1, PEPCK and G-6-P were lower by 26.6%, 35.0%, 34.3% (P less than 0.01). Meanwhile, liver steatosis was also milder.

CONCLUSIONCalorie restriction can inhibit the expressions of FoxO1, PEPCK and G-6-P, strengthen insulin signal conduction, suppress gluconeogenesis and thus regulate glycometabolism.

Animals ; Caloric Restriction ; Dietary Fats ; Forkhead Transcription Factors ; genetics ; Gene Expression Regulation ; Gluconeogenesis ; genetics ; Glucose-6-Phosphatase ; genetics ; Liver ; metabolism ; Male ; Nerve Tissue Proteins ; genetics ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Rats ; Rats, Wistar

To develop a method for the detection of surface-confined peptides containing cysteine residues or oligodeoxynucleotides (ODNs) whose 3' ends modified with thiol groups, and a thiol-specific fluorescent cross-linker, N-(9-acridinyl) maleimide (NAM) was used. The peptides studied herein include both the oxidized and reduced forms of glutathione, and a hexapeptide (FT). Peptides are first attached onto the activated 11-mercaptoundecanoic acid (MUA)-terminated alkanethiol self-assembled monolayers (SAMs) and then derivatized with NAM. The cysteine residues was determined by using electrochemical desorption and fluorescence detection. GSH concentration as low as 40 pmol x L(-1) can be measured. The fluorescence intensity in the case of FT is about 3 times as high as that for GSH, which is consistent with the molar ratio of cysteine residues in these two molecules. The analytical performance of gene analysis was also evaluated through the analyses of a complementary target and targets with varying numbers of mismatching bases. The method described here is simple, sensitive, reproducible, and does not require sophisticated analytical instrumentation and separation procedures.
Biosensing Techniques ; methods ; Cysteine ; analysis ; Electrochemistry ; methods ; Fluorescence ; Glutathione ; analysis ; chemistry ; Maleimides ; chemistry ; Oligodeoxyribonucleotides ; analysis ; Reproducibility of Results ; Sensitivity and Specificity

Objective:To establish the best evidence-based approach for early fluid resuscitation management in patients with severe acute pancreatitis (SAP).Methods:A literature search was conducted utilizing evidence-based nursing methods to identify relevant evidence on the management of early fluid resuscitation in patients with SAP. The search followed the hierarchical order of the " 6S" evidence pyramid, including databases such as China National Knowledge Infrastructure (CNKI), China Biomedical Literature Database (SinoMed), Wanfang Database, UpToDate, NICE, RNAO Guidelines Network, Pancreatology International, WHO Association Website, JBI, Cochrane, PubMed, EMBASE, and CINAHL. The search was limited to articles published from the establishment of each database to March 2022. The literature quality evaluation tools and an evidence pre-grading system from the JBI Evidence-Based Health Care Center were employed to assess the quality of the literature included in the study. Additionally, the FAME structure was utilized to evaluate the feasibility, appropriateness, clinical significance, and validity of the evidence.Results:Nine articles were finally incorporated into the analysis, including four guidelines, one evidence summary, two systematic reviews, and two expert consensus articles. 21 pieces of evidence pertaining to early fluid resuscitation management in patients with SAP was summarized, encompassing five key aspects: resuscitation timing, type of fluid infusion, total volume and speed of fluid infusion, dynamic monitoring, and fluid resuscitation goals. It was advisable for patients diagnosed with SAP to promptly receive fluid resuscitation, ideally within 72 hours of diagnosis. The initial choice for fluid resuscitation was lactated Ringer′s solution, with the addition of human albumin as a supplementary colloid solution. The quantity of fluid administered within the first 24 hours of rehydration should constitute approximately 33.3% of the total rehydration volume within the 72 hours time-frame. In the case of patients experiencing early shock or dehydration, it was advised that the fluid rate administered should be 5-10 ml·kg -1·h -1 within the first 24 hours of admission. Additionally, an infusion of 20 ml/kg of fluid can be administered within the initial 30-45 minutes. It was recommended to assess the adequacy of early fluid resuscitation every 4-6 hours, ensuring that the resuscitation objective could meet at least two of the following criteria: urine output of 0.5-1 ml·kg -1·h -1, mean arterial pressure of 65-85 mmHg, central venous pressure of 8-12 mmHg, heart rate below 120 beats/min, central venous oxygen saturation of at least 70%, and a decrease in hematocrit levels to 30%-35%. Conclusions:The most compelling evidence supporting the implementation of early fluid resuscitation management in patients with SAP is derived from an evidence-based nursing approach, which could effectively improve patient care outcomes.

4-(Cytidine-5-diphospho) -2-C-methyl-D-erythritol kinase is a key enzyme in the biosynthesis pathway of terpenoids. According to the transcriptome database, the specific primers were designed and used in PCR. The bioinformatic analysis of the sequenced TwCMK gene was performed in several bioinformatics software. The Real-time fluorescence quantification polymerase chain reaction (RT-qPCR) were used to detect the expression levels of TwCMK from T. wilfordii after elicitor MeJA supplied. The results showed that the full length of TwCMK cDNA was 1 732 bp encoding 387 amino acids. The theoretical isoelectric point of the putative TwCMK protein was 5.79 and the molecular weight was about 42.85 kDa. MeJA stimulated the rising of TwCMK expression in suspension cell and signally impacted at 24 h. The research provides a basis for further study on the regulation of terpenoid secondary metabolism and biological synthesis.
Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; Gene Expression Regulation, Plant ; Models, Molecular ; Molecular Sequence Data ; Phosphotransferases (Alcohol Group Acceptor) ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Tripterygium ; chemistry ; enzymology ; genetics

Seashore landfill aquifers are environments of special physicochemical conditions (high organic load and high salinity), and microbes in leachate-polluted aquifers play a significant role for intrinsic bioremediation. In order to characterize microbial diversity and look for clues on the relationship between microbial community structure and hydrochemistry, a culture-independent examination of a typical groundwater sample obtained from a seashore landfill was conducted by sequence analysis of 16S rDNA clone library. Two sets of universal 16S rDNA primers were used to amplify DNA extracted from the groundwater so that problems arising from primer efficiency and specificity could be reduced. Of 74 clones randomly selected from the libraries, 30 contained unique sequences whose analysis showed that the majority of them belonged to bacteria (95.9%), with Proteobacteria (63.5%) being the dominant division. One archaeal sequence and one eukaryotic sequence were found as well. Bacterial sequences belonging to the following phylogenic groups were identified: Bacteroidetes (20.3%), beta, gamma, delta and epsilon-subdivisions of Proteobacteria (47.3%, 9.5%, 5.4% and 1.3%, respectively), Firmicutes (1.4%), Actinobacteria (2.7%), Cyanobacteria (2.7%). The percentages of Proteobacteria and Bacteroides in seawater were greater than those in the groundwater from a non-seashore landfill, indicating a possible influence of seawater. Quite a few sequences had close relatives in marine or hypersaline environments. Many sequences showed affiliations with microbes involved in anaerobic fermentation. The remarkable abundance of sequences related to (per)chlorate-reducing bacteria (ClRB) in the groundwater was significant and worthy of further study.
Bacteria ; genetics ; isolation & purification ; Environmental Monitoring ; methods ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; analysis ; genetics ; Seawater ; analysis ; microbiology ; Sewage ; microbiology ; Water Microbiology ; Water Pollutants ; analysis

OBJECTIVETo investigate the differentially expressed genes in rat in the process of regression of vascular calcification by using the suppression subtractive hybridization (SSH).

METHODS24 SD male rats which aged 6 weeks and specific pathogen free grade were selected and randomly divided into 3 groups (n = 8): control group, calcification group and regression group respectively. Vascular calcification model (vitamin D3 plus nicotine, VDN) were made from rats in calcification group and regression group, and rats in control group were intragastric administered with normal saline and lavaged with peanut oil. Rats were bred for 8 weeks in calcification group and control group, while rats in regression group were fed for 16 weeks. All rats were killed to measure concentration of calcium in the arterial tissue and examine the pathological lesion changes. Subtractive hybridization among vascular cDNA sequences from calcification group and regression group were established. The cDNA fragments which expressed higher or lower in regression group than those in calcification group were isolated. Differentially expressed genes with cDNA fragment were inserted into PMD18-T plasmid vector and transformed competent DH-5alpha, cDNA libraries of differentially expressed gene between calcification group and regression group were then constructed. Recombinant vectors were analyzed by colony PCR, positive genes were randomly selected for sequencing and analyzed by BLAST. 4 genes were randomly selected for RT-PCR certification combined with semi-quantitative analysis of DNA bands.

RESULTSVDN model of rats were successfully constructed. Concentration of tissue calcium in calcification group (15.34 mg/g +/- 2.51 mg/g) was significantly increased compared to that in control group (5.20 mg/g +/- 0.75 mg/g, P < 0.001), while in comparison with calcification group (15.34 mg/g +/- 2.51 mg/g), calcium in regression group was relatively lower (12.73 mg/g +/- 1.89 mg/g, P < 0.05). 28 up-regulated genes and 22 down-regulated genes were gained through sequencing and BLAST analysis among positive clones. RT-PCR validation indicated that 4 genes such as prdx3 and Ank2 had increasedly expressed in regression group than those in calcification group, the average fold change was 1.7.

CONCLUSIONRat vascular calcification tissue had characteristic of active regression. Genes in relation to pyrophosphoric acid synthesis, glutamate signal peptides, anti-oxidant and ant-apoptosis were up-regulated, at the same time many genes related to ossification and oxidation activity were down-regulated in the process of calcification regression. Increased expression of calcification suppressor genes accompanying decreased expression of calcification promoting genes might be the intrinsic mechanisms which initiated the active regression of calcified tissues.

Animals ; Aorta ; metabolism ; pathology ; Gene Expression Profiling ; Gene Expression Regulation ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Calcification ; genetics ; physiopathology

OBJECTIVETo study the prevalence of primary HIV drug resistance in antiretroviral therapy (ART) areas of Henan province.

METHODSA total of 121 drug-naive long-term infected individuals and 154 patients with newly diagnosed from January 2011 to March 2012 were recruited, the questionnaires were surveyed and whole blood were collected to analyze the CD4(+)T cell counts and viral load. In-house method for genotypic resistance test was determined in those with viral load > 1000 copies/ml samples, the differences of demographic characteristics, immunological parameters and primary drug resistance were compared between the two groups.

RESULTSA total of 121 cases of long-term individuals who had infected (12.50 ± 3.21) years were mainly previous paid blood donors, and the age was (46.61 ± 9.32) years old. The infection route of the newly diagnosed were diversity, including blood, sexual transmission and others, the cases were 73, 73, 8, respectively, the confirmatory year was (0.91 ± 0.28) years, and average age was (22.21 ± 3.11) years old. The difference were statistically significant in the route of transmission, age and infection time from demographic analysis of the two groups (P < 0.05). The absolute M(P(25)-P(75)) counts of CD4(+)T lymphocytes of long-term group was 322 (217 - 422) cell/µl, which was lower than the newly diagnosed was 434(308 - 578) cell/µl (P < 0.05), and viral load was 4.0 (2.96 - 4.64) copies/ml, 3.77 (2.94 - 4.53) copies/ml, the difference was not significant (P > 0.05). The prevalence of primary drug resistance in long-term group and newly diagnosed was 5.79% (7/121), 9.09% (14/154), respectively, and the difference was statistically different (P < 0.05), and one PI-resistant strain was found in the newly diagnosed group.

CONCLUSIONThe primary drug resistant strains in untreated patients were found in Henan province of ART areas, and there was difference in degree of resistance between long-term infected individuals and newly diagnosed.

Adult ; Anti-HIV Agents ; pharmacology ; therapeutic use ; China ; epidemiology ; Drug Resistance, Viral ; Female ; HIV Infections ; drug therapy ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Viral Load

BACKGROUNDEndothelial dysfunction is considered as the initiating process and pathological basis of cardiovascular disease. Cyclooxygenase-2 (COX-2) and prostacyclin synthase (PGIS), inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) are key enzymes with opposing actions in inflammation and oxidative stress, which are believed to be the major driver of endothelial dysfunction. And in hypoxia (Hx), Hx-inducible factor (HIF)-1α and HIF-2α are predominantly induced to activate vascular endothelial growth factor (VEGF), resulting in abnormal proliferation. Whether and how Tongxinluo (TXL) modulates COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α, and VEGF in Hx-stimulated human cardiac microvascular endothelial cells (HCMECs) have not been clarified.

METHODSHCMEC were treated with CoCl 2 to mimic Hx and the mRNA expressions of COX-2, PGIS, iNOS, eNOS, HIF-1α, HIF-2α, and VEGF were first confirmed, and then their mRNA expression and protein content as well as the cell pathological alterations were evaluated for TXL treatment with different concentrations. In addition, the effector molecular of inflammation prostaglandin E 2 (PGE 2 ) and the oxidative marker nitrotyrosine (NT) was adopted to reflect HCMEC injury.

RESULTSHx could induce time-dependent increase of COX-2, iNOS, HIF-2α, and VEGF in HCMEC. Based on the Hx-induced increase, TXL could mainly decrease COX-2, iNOS, HIF-2α, and VEGF in a concentration-dependent manner, with limited effect on the increase of PGIS and eNOS. Their protein contents verified the mRNA expression changes, which was consistent with the cell morphological alterations. Furthermore, high dose TXL could inhibit the Hx-induced increase of PGE 2 and NT contents, attenuating the inflammatory and oxidative injury.

CONCLUSIONSTXL could inhibit inflammation-related COX-2, oxidative stress-related iNOS, and HIF-2α/VEGF to antagonize Hx-induced HCMEC injury.

Blotting, Western ; Cell Hypoxia ; drug effects ; Cell Line ; Cobalt ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism