Aged, 80 and over ; Blood Vessel Prosthesis Implantation ; adverse effects ; methods ; Humans ; Male ; Pacemaker, Artificial ; Subclavian Artery ; surgery

Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca~(2+)]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca~(2+)]i was examined using confocal laser scanning microscopy (CLSM) and the Ca~(2+) sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca~(2+)]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca~(2+)]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca~(2+) occurred,which resulted in an increase of [Ca~(2+)]i.Extracellular bound calcium was therefore converted into intracellular free calcium.

Objective To investigate the radiosensitization effects of the combination treatment of clioquinol (CQ) and zinc on human cervical cell line HeLa in vitro.Methods Cells were divided into the 4 groups:controls,drug,radiation,and combined drug and radiation group.Cytotoxic effect of CQ and zinc on cell viability was determined by CCK-8 assay.Radiosensitization effect of CQ and zinc on HeLa cells was detected by colongenic assay,and the single-hit multi-target model was used to stimulate the doseresponse curve of survival and to calculate radiosensitization parameters.The cell cycle and apoptosis of HeLa cells were analyzed with flow cytometry.Luciferase reporter assay was used to study NF-κB activity of HeLa cells.Results The combination of CQ and zinc inhibited cell growth in a dose-dependent manner (F =188.00,P ＜ 0.01).The mean lethal dose was 3.16 and 2.04 Gy for radiation group and combined drug and radiation group,respectively,and hence the SER was 1.55.Compared with the radiation group,the ratio of G2-phase cells in the combined drug and radiation group decreased(t =10.39,P ＜ 0.05),the apoptosis rate increased at 24 h post-irradiation (t =5.64,P ＜ 0.01),and the NF-κB activity decreased (t =21.42,P ＜ 0.05).Compared to the control group,the NF-κB activity increased in the radiation group(t=6.23,P＜0.05),but decreased in the drug group(t =12.48,P＜0.05).Conclusions The combination of CQ and zinc could increase the radiosensitivity of HeLa cells by decreasing the ratio of G2-phase cells,increasing apoptosis and the inhibiting of NF-κB activity.

Objective To investigate the effect of early low-glucocorticoid on hemodynamics and prognosis in the patients with septic shock.Methods Sixty patients with septic shock failing in active fluid resuscitation and vasoactive drugs in our hospital from June 2013 to August 2015 were selected and divided into the control group,early-hormone group and late-hormone group.MAP,HR,PO2/FIO2 and serum lactic acid levels were monitored in all selected patients before treatment and at 12,24,48 h after treatment.Apache Ⅱ,SOFA scores were assessed before treatment and on 1,3,7 d after treatment.The ventilation time,ICU stay time,hospital stay time and intravenous use time of vasoactive agents(VDNT) were recorded.Results The Apache Ⅱ scores and SOFA scores on 3,7 d after treatment in the early-hormone group were significantly decreased compared with the late-hormone group and control group (P＜0.05).MAP and HR at 24,48 h after treatment in the early-hormone group were significantly improved compared with the late-hormone group and control group (P＜0.05).The level of serum lactic acid at 12,24 h after treatment in the early-hormone group and late-hormone group were obviously lower than that in the control group,the levels of serum lactic acid at 12,24 h after treatment in the early-hormone group were obviously lower than those in the late-hormone group (P＜ 0.05).PO2/FIO2 at 12 h after treatment in the early-hormone group and late-hormone group were obviously better than that in the control group,and PO2/FIO2 at 12 h after treatment in the early-hormone group was obviously better than that in the late-hormone group(P＜0.05).The ventilation time,ICU stay time,hospital stay time and VDUT in the early-hormone group were significantly shortened compared with the late-hormone group and control group.The ventilation times,ICU stay time and VDUT in the latehormone group were significantly shortened compared with the control group (P＜0.05).Conclusion Early using low-dose glucocorticoid may restore hemodynamics more quickly,protects the organ function and improves the prognosis in the patients with septic shock.

Objective: To analyze the mortality trend of bladder cancer among residents in Qidong, Jiangsu Province from 1972 to 2016, so as to provide the basis for the prevention and treatment strategy of bladder cancer in Qidong. Methods: The data of bladder cancer was collected from Qidong Cancer Registry.The crude mortality rate ( CR ), age-standardized rate by Chinese population in 2000 (CASR) and world population in 1960 ( WASR ), truncated rate (35-64 years) and cumulative rate ( 0-74 years ) were calculated. The annual percent change ( APC ) was used to analyze the trend of mortality in bladder cancer. Results: During from 1972 to 2016, There were 1 497 deaths due to bladder cancer in Qidong from 1972 to 2016. The CR, CASR and WASR were 2.96/105, 1.83/105 and 1.80/105, respectively. The APCs in CR, CASR, WASR of bladder cancer were 5.29%, 1.86% and 1.81%, respectively ( P<0.05 ), showing upward trends. The truncated rate, cumulative rate and cumulative risk were 1.47/105, 0.17% and 0.17%, respectively. The CR, CASR and WASR in males were 4.71/105, 2.97/105 and 3.31/105, respectively, which was higher than that of 1.26/105, 0.75/105, and 0.66/105 in females ( P<0.05 ). The APC of CR, CASR and WASR in males were 5.71%, 1.96% and 2.17%, respectively ( P<0.05 ), all showed upward trends. For females, the APC of CR was 4.47% ( P<0.05 ), showing an upward trend, but there was no significant change in CASR and WASR ( P>0.05 ). The CR of bladder cancer was high among people aged more than 55 years. The CR in 55-64-year-old group, 65-74-year-old group and more than 75-year-old group showed upward trends, with APC of 4.50%, 2.22% and 4.51%, respectively ( P<0.05 ). Conclusions From 1972 to 2016, the mortality of bladder cancer in Qidong showed an upward trend, which was relatively high in men and people aged over 55 years.

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

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.

OBJECTIVEStudying the gene expression profiling of different stage hairy root of Salvia miltiorrhiza, in order to find functional genes.

METHODThe contents of second metabolites were determined by HPLC and gene expression profiling was detected by cDNA microarray. cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine's linear RNA amplification method and subsequent enzymatic reaction. The microarrays were scanned with a ScanArray Express scanner using ScanArray 2.0 software and quantified by signal intensities of individual spots from the 16-bit TIFF images using GenePix Pro 4.0. The linear normalization method was used for data analyze. Northern blot was used to test the gene expression results obtained by microarray. Different expressed genes were sequenced and analyzed by gap4 software, and then they were analyzed with BLASTX, BLASTN, GO and KEGG.

RESULTGrowth rate and second metabolites analysis indicated that the stage from 30 d to 45 d was the growth stage, while the stage from 45 d to 60 d was the second metabolites accumulation stage. Accordingly 30 d hairy root was chosen as a reference, which was hybridized with 45 d and 60 d hairy root separately. Total 203 different expressed genes were obtained. Northern blot showed that the result was identical with the microarray result. After sequenced, there were 172 genes clustered into 114 clusters (Unigenes). Among them, 62 unigenes had known functions, 34 unigenes were hypothetical protein, 9 unigenes were homologues with no similarity and 9 unigenes were unidentified protein with low similarity. Total 67 genes were classified into cellular component ontology, molecular function ontology and biological process ontology based on GO analysis. Total 26 genes, which represented 29 metabolic-related enzymes, were located in metabolic maps based on KEGG pathway classification.

CONCLUSIONSeveral important functional genes related to second metabolite synthesis were cloned such as P450 and copalyl diphosphate synthase genes. cDNA microarray was a useful tool for functional genomics of traditional Chinese medicine.

Alkyl and Aryl Transferases ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism

OBJECTIVETo study on the chemical stability of Salvia miltirrhiza hairy root.

METHODThe rolA gene was detected by PCR in DNA and the chemical contituent variances were detected by HPLC.

RESULTThe rolA gene was found in all the 10 batches of the culfured hairy root. The similarities of the chromatographic fingerprints of the 10 batches are higher than 0. 95.

CONCLUSIONThere are no significant differences of the chemical constituents in 10 hairy root samples.

Chromatography, High Pressure Liquid ; DNA, Plant ; genetics ; Drugs, Chinese Herbal ; chemistry ; Genes, Plant ; Plant Roots ; chemistry ; genetics ; Polymerase Chain Reaction ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; genetics ; Sensitivity and Specificity

OBJECTIVETo investigate the effects of excessive intake of fluoride on the expression of type II collagen gene and types and morphological change of collagen fiber in the bone tissues of rats.

METHODSA rat model with fluorosis was established by adding 221 mg/L of sodium fluoride (NaF) to drinking water for the rats for 15 days, 30 days and two months, respectively. Type II collagen alpha1 (II) cDNA probe was prepared, and cDNA-mRNA in-situ hybridization was employed to detect change in expression of type II collagen mRNA in the bone tissues of rats with excessive intake of fluoride (221 mg/L NaF). Picrosirius-polarization method was used to observe types of collagen and morphology of collagen fiber in the bone tissues.

RESULTSChondroblasts were found in the femur and other bone tissues of the rats after exposure to fluoride. cDNA-mRNA in-situ hybridization showed that expression of type II collagen gene could be observed in the cytoplasm of chondrocytic lacuna and chondrified bone tissues. mRNA in collagen of chondrocytes of the rib cartilage reached the peak level 15 days after exposure to fluoride, and decreased gradually one month and two months after exposure. Polychromatic type II collagen, breakage of collagen fiber, disorder array and reduced content of type II collagen could be found in the bone tissues with picrosirius-polarization method.

CONCLUSIONSExcessive intake of fluoride could lead to changes in types and structure of collagen (cross-linkage) of bone tissues, which caused expression of type II collagen gene in the chondrified bone tissues and enhanced its expression in the rib cartilage tissues.

Animals ; Bone Diseases ; metabolism ; pathology ; Chondrocytes ; metabolism ; Collagen Type II ; biosynthesis ; genetics ; Fluoride Poisoning ; genetics ; metabolism ; pathology ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar